Phytochemical, Vitamin, and Actives testing

This assay quantifies 10-Hydroxy-2-decenoic acid (10-HDA), the primary fatty acid unique to royal jelly. Using HPLC, it verifies 10-HDA concentration in raw royal jelly, powders, and finished products to confirm authenticity, potency, and compliance with pharmacopeial quality standards.

Samples are extracted with methanol or other suitable organic solvents and analyzed by HPLC with UV detection (typically at ~215 nm). Quantification is performed using certified 10-HDA standards, with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in mg/g or % w/w of 10-HDA. Values are compared against specification targets (e.g., ≥1.4% 10-HDA in fresh royal jelly) and label claims to confirm product authenticity, detect adulteration, and verify consistency across batches.

This test quantifies 3’,3’-Diindolylmethane (DIM), a bioactive compound derived from indole-3-carbinol, important for assessing product potency and quality. The HPLC method accurately measures DIM levels in raw materials, powders, and finished dietary supplements. Results are reported in mg per gram or per serving, with a detection limit suitable for low-level quantification.

Samples are prepared by extracting 0.5 g of material with methanol using sonication for 30 minutes, followed by filtration. The extract is analyzed using High-Performance Liquid Chromatography equipped with a UV detector set at 280 nm. DIM is separated on a C18 reversed-phase column with a gradient mobile phase of water and acetonitrile. Quantification is performed against a certified DIM reference standard using a five-point calibration curve. Method accuracy is verified through duplicate injections, spiked recovery tests, and analysis of quality control samples.

Results are reported in mg/g (raw material) or mg/serving (finished products). Testing verifies standardized potency, supports label claims, and ensures consistency in botanical and nutraceutical formulations.

This assay quantifies 5-hydroxytryptophan (5-HTP), a serotonin precursor derived from griffonia seed extract, in dietary supplements. Using HPLC, it verifies the concentration of 5-HTP to confirm label claims and ensure consistent dosing in mood and relaxation products.

Samples are extracted in an acidified aqueous solution, then analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified 5-HTP standards, with internal standard correction and duplicate runs to ensure accuracy.

Results are reported in mg per g or per serving. Values are assessed against formulation targets and label declarations to ensure potency, quality, and stability throughout shelf life.

This test identifies and quantifies 5′-methylthioadenosine (MTA), also known as methylthioadenosine, in raw materials and finished dietary supplement products using High-Performance Liquid Chromatography (HPLC) with liquid chromatography–mass spectrometric (LC-MS) confirmation or quantitation, as appropriate to the matrix and reporting requirement. MTA is a naturally occurring sulfur-containing nucleoside formed in cellular methylation and polyamine-biosynthesis pathways. It may occur in fermentation-derived materials and in certain processed botanical matrices, including aged garlic preparations. Because MTA concentrations can vary with source material, cultivation, processing, fermentation, and storage conditions, quantitative testing is useful for raw-material characterization, specification control, lot-to-lot consistency, and substantiation of any accurately stated MTA-content claim. Results are reported as mg/g, mg/kg, or mg per serving, as specified by the product format and client requirement.

A representative sample is accurately weighed and extracted using a validated aqueous or aqueous-organic solvent system, commonly water, dilute acidified water, or aqueous methanol, selected to recover the polar nucleoside while minimizing co-extracted matrix interference. The extract is clarified by centrifugation and membrane filtration before chromatographic analysis. MTA is separated by reversed-phase HPLC, typically using a C18 column and an aqueous mobile phase with a volatile buffer or acid modifier compatible with both UV and mass-spectrometric detection. HPLC-UV detection is generally performed near the adenine chromophore absorption maximum (commonly approximately 260 nm). LC-MS or LC-MS/MS provides confirmatory molecular specificity through retention-time agreement and the characteristic ionization and fragmentation behavior of MTA; it may also serve as the primary quantitation technique when enhanced selectivity or sensitivity is required. Quantification is performed against a multi-point external calibration curve prepared from a qualified MTA reference standard, with standard traceability, purity correction, and calibration acceptance documented. Method blanks, duplicate preparations, matrix spikes or recovery controls, and quality-control samples are included to verify accuracy, precision, linearity, and freedom from material matrix interference

MTA is a polar, UV-absorbing adenine nucleoside that is well suited to chromatographic separation and quantification. HPLC provides efficient resolution of MTA from other nucleosides, amino acids, sugars, sulfur compounds, and formulation excipients, while LC-MS confirmation adds molecular selectivity that is especially valuable for complex matrices such as aged garlic, fermented powders, multicomponent botanicals, and finished supplements. The combined HPLC/LC-MS approach reduces the risk of assigning an MTA result to co-eluting UV-active compounds and provides defensible confirmation of both identity and concentration. This method supports establishment and verification of ingredient specifications, label-claim substantiation where applicable, and dietary supplement cGMP quality-control requirements under 21 CFR 111.

This test quantifies 6-shogaol — the principal dehydration product of 6-gingerol and the most abundant shogaol homolog in dried and heat-processed ginger (Zingiber officinale Roscoe) — in raw materials, dried ginger powders, extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). 6-Shogaol is formed from 6-gingerol through dehydration of the β-hydroxyl group during drying, heating, or prolonged storage of ginger, and is present at substantially higher concentrations in dried ginger and processed extracts than in fresh rhizome. Emerging research indicates that 6-shogaol exhibits potent antioxidant, anti-inflammatory, and neuroprotective activities that in several models exceed those of 6-gingerol, making its specific quantification increasingly important for characterizing the full bioactive profile of ginger materials. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and extracted using an appropriate organic solvent system (e.g., methanol or aqueous methanol) to ensure complete extraction of 6-shogaol and co-occurring ginger phenylalkylketones. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 282 nm, the characteristic absorption maximum of the phenylalkylketone chromophore of 6-shogaol. Chromatographic conditions are optimized to achieve baseline resolution of 6-shogaol from co-eluting gingerol and shogaol homologs, particularly 6-gingerol and 8-shogaol, which have similar retention characteristics. Quantification is performed against a multi-point external calibration curve prepared from a certified 6-shogaol reference standard. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

6-Shogaol and 6-gingerol share the same phenylalkylketone chromophore and similar chromatographic behavior, making their specific resolution and individual quantification by HPLC essential to ensure that potency results accurately reflect 6-shogaol content rather than total pungent compound content. The ratio of 6-shogaol to 6-gingerol is also a meaningful quality indicator of ginger processing history, with high shogaol-to-gingerol ratios characteristic of dried or heat-processed materials. Specific 6-shogaol quantification supports label claim accuracy for ginger extracts standardized to shogaol content, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay measures 7,8-dihydroxyflavone, a flavonoid compound, using High-Performance Liquid Chromatography (HPLC). The analysis provides accurate and selective quantification for quality control and standardization.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. 7,8-Dihydroxyflavone is separated from related flavonoids and detected via UV or diode-array detection. Quantitation is performed using certified reference standards with calibration curves and replicate injections ensuring accuracy and reproducibility.

Testing verifies standardized potency, confirms label claims, and ensures batch-to-batch consistency.

This test confirms the identity of acacia gum (gum arabic; Acacia senegal (L.) Willd., syn. Vachellia senegal, and Acacia seyal Del.) in raw materials, spray-dried powders, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Acacia gum is a complex, branched polysaccharide exudate derived from the stems and branches of Acacia senegal and related species, composed primarily of arabinogalactan with associated glycoproteins. It is widely used in dietary supplements as a prebiotic soluble fiber, encapsulation matrix, and excipient, and is recognized as a Generally Recognized as Safe (GRAS) ingredient by the FDA. HPTLC identity testing employs acid hydrolysis to release the characteristic monosaccharide constituents — primarily arabinose, galactose, rhamnose, and glucuronic acid — which are resolved and visualized as a diagnostic sugar fingerprint compared against an authenticated acacia gum reference standard to confirm identity and detect potential adulteration or substitution with other gum exudates or polysaccharide materials.

A representative sample is accurately weighed and subjected to complete acid hydrolysis (e.g., using dilute sulfuric or trifluoroacetic acid under reflux or sealed-vessel conditions) to hydrolyze the arabinogalactan polysaccharide into its constituent monosaccharides. The hydrolysate is neutralized, filtered, and applied alongside an authenticated acacia gum hydrolysate reference standard, individual monosaccharide reference standards (arabinose, galactose, rhamnose, glucuronic acid), and, where applicable, potential adulterant hydrolysates (e.g., gum ghatti, gum karaya, guar gum), onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated aqueous-organic solvent system optimized for monosaccharide resolution. After development, the plate is derivatized with an appropriate sugar-visualizing reagent (e.g., diphenylamine-aniline-phosphoric acid or anisaldehyde-sulfuric acid) and evaluated under white light and UV light. The resulting monosaccharide fingerprint — characterized by the characteristic arabinose, galactose, rhamnose, and glucuronic acid band pattern diagnostic of acacia arabinogalactan — is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Acacia gum is subject to adulteration and substitution with other gum exudates — including gum ghatti, gum karaya, gum tragacanth, and mesquite gum — as well as lower-cost polysaccharide fillers that may be visually indistinguishable in dried powder form. HPTLC monosaccharide fingerprinting following acid hydrolysis provides a practical and discriminating identity confirmation method that exploits the characteristic monosaccharide composition of acacia arabinogalactan — notably the co-presence of arabinose, galactose, rhamnose, and glucuronic acid in characteristic proportions — to distinguish authentic acacia gum from other gum exudates with different monosaccharide profiles. This method aligns with USP and Food Chemicals Codex (FCC) identity testing guidelines for acacia gum and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This assay quantifies acetic acid using High-Performance Liquid Chromatography (HPLC). The method provides accurate, selective determination of volatile organic acids across diverse matrices such as vinegar, kombucha, and fermented products.

Samples are filtered and injected into an HPLC system equipped with an organic acid or reverse-phase column. Acetic acid is separated and detected by UV or refractive index (RI) detection, quantified against certified reference standards. Duplicate runs and calibration curves ensure precision and reproducibility.

Results are reported in mg/mL (liquids) or mg/g (solids). Testing verifies fermentation quality, confirms product specifications, and ensures consistency across production batches.

This test quantifies adenosine in dietary supplements, botanical extracts, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Adenosine is a naturally occurring nucleoside found in mushroom extracts such as Cordyceps and in other botanical ingredients, where it serves as a key bioactive marker compound. It plays a fundamental role in cellular energy transfer, cardiovascular regulation, and neurotransmission. Results are reported in mg per serving or mg per gram to support label claim verification, standardization of botanical extracts, and cGMP compliance.

A representative sample is weighed and extracted using a methanol-water solvent system with sonication to ensure complete solubilization of adenosine from the matrix. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 260 nm, which corresponds to the characteristic UV absorbance of the purine ring system. Quantification is performed against a multi-point external calibration curve prepared from a certified adenosine reference standard, with system suitability and QC samples run concurrently to confirm method performance.

Adenosine is the primary bioactive marker used to standardize Cordyceps and related mushroom extracts, and its accurate quantification is essential for verifying extract potency and label claim compliance. HPLC-UV at 260 nm provides the selectivity needed to resolve adenosine from other nucleosides and co-extractives present in complex botanical matrices, delivering reliable and reproducible results for both raw material qualification and finished product release testing.

This assay quantifies agmatine sulfate, a metabolite of arginine commonly used in pre-workout and neuro-support supplements. Using LC-MS/MS, it verifies agmatine content to ensure correct dosing, label accuracy, and formulation integrity in sports and cognitive performance products.

Samples are extracted in aqueous or acidified solution and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed with certified agmatine standards, internal standard correction, and duplicate injections to ensure precision.

Results are reported in mg per g or per serving. Values are compared to formulation targets and declared label claims. Testing ensures proper inclusion and helps detect underdosing or degradation during manufacturing or storage.

This test quantifies 3-acetyl-11-keto-beta-Boswellic Acid (AKBA), a key anti-inflammatory compound found in Boswellia serrata extracts. Using High-Performance Liquid Chromatography (HPLC), the assay verifies that raw materials, powders, and finished products meet specified AKBA concentration levels, typically reported in mg/g. Accurate measurement of AKBA ensures product efficacy and compliance with quality standards.

Samples are first extracted with methanol to solubilize AKBA from the matrix. The extract is filtered and injected into an HPLC system equipped with a reversed-phase C18 column. Detection is performed using UV absorbance at 250 nm, where AKBA exhibits a characteristic peak. Quantification is achieved by comparing sample peak areas to a calibration curve constructed from certified AKBA reference standards. Method accuracy is confirmed through duplicate injections, inclusion of quality control samples, and spike recovery tests.

Results are reported in % w/w (raw material) or mg/serving (finished products). Testing verifies standardized potency, supports label claims, and ensures consistent quality of Boswellia-based supplements.

This test quantifies alliin (S-allyl-L-cysteine sulfoxide) — the principal sulfur-containing amino acid derivative and primary bioactive precursor compound in garlic (Allium sativum L.) — in raw materials, garlic powders, aged garlic extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Alliin is the stable, odorless precursor to allicin (diallyl thiosulfinate), which is enzymatically generated from alliin by the enzyme alliinase upon cell disruption. Alliin content is the primary standardization marker for garlic powder and extract ingredients, as it directly determines the allicin-generating potential (allicin yield) of the material. Accurate alliin quantification is essential for label claim substantiation, raw material qualification, and for predicting the allicin release capacity of garlic-based products. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and immediately dissolved in an appropriate acidic aqueous solvent (e.g., dilute hydrochloric acid or acidified methanol) to inhibit alliinase enzyme activity and prevent enzymatic conversion of alliin to allicin during sample preparation. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 210 nm, as alliin lacks a strong chromophore at higher UV wavelengths and is best detected at low UV wavelengths where the sulfoxide functional group and amino acid backbone provide adequate absorbance. Quantification is performed against a multi-point external calibration curve prepared from a certified alliin reference standard. Where applicable, related organosulfur compounds — including S-allylcysteine (SAC), S-methylcysteine sulfoxide, and γ-glutamyl-S-allylcysteine — may be monitored simultaneously to provide a broader organosulfur profile. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Accurate alliin quantification requires careful control of sample preparation conditions to prevent enzymatic conversion of alliin to allicin by endogenous alliinase, which is rapidly activated upon cell disruption and aqueous contact. Acidification of the extraction solvent effectively inhibits alliinase activity, ensuring that alliin is measured in its intact precursor form. HPLC with UV detection at 210 nm provides the sensitivity required for alliin quantification in the absence of a strong chromophore, with chromatographic separation resolving alliin from co-occurring organosulfur amino acids and matrix components. Alliin content is the internationally recognized potency specification for garlic powder and extract ingredients, referenced in the European Pharmacopoeia (Ph. Eur.) monograph for garlic powder, and supports label claim substantiation and cGMP compliance under 21 CFR 111.

This assay measures allulose (D-psicose), a rare monosaccharide used as a low-calorie sweetener, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high specificity and sensitivity for accurate quantification in complex food and supplement matrices.

This assay measures allulose (D-psicose), a rare monosaccharide used as a low-calorie sweetener, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high specificity and sensitivity for accurate quantification in complex food and supplement matrices.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Allulose is detected using multiple reaction monitoring (MRM) and quantified against certified reference standards. Internal calibration and quality control checks ensure precision and reproducibility.

This assay quantifies aloin, a naturally occurring anthraquinone glycoside found in Aloe vera latex. Aloin levels are regulated due to its laxative properties and potential safety concerns at high concentrations.

Samples are extracted and analyzed by High-Performance Liquid Chromatography (HPLC) with UV detection. Aloin A and B are separated, detected, and quantified against certified standards. Method validation includes calibration curves and quality control checks.

Results are reported in ppm (mg/kg) for raw materials or mg/serving for finished products. Monitoring aloin ensures compliance with safety guidelines (e.g., USP/EFSA limits), verifies raw material purity, and supports accurate labeling.

This assay quantifies Alpha-GPC (Alpha-Glycerylphosphorylcholine), a highly bioavailable choline donor used in nootropic and cognitive health supplements. Using LC-MS/MS, it verifies Alpha-GPC content in capsules, powders, and functional blends to confirm potency and ensure label compliance.

Samples are extracted using aqueous or methanolic solvents and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified Alpha-GPC standards, with internal standard correction and duplicate injections to ensure precision and reproducibility.

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm dosing accuracy and detect degradation or adulteration.

This assay determines the complete profile of individual amino acids present in a food sample following protein hydrolysis. The process breaks down proteins into their constituent amino acids, which are then separated and quantified. This profile is essential for evaluating nutritional quality and verifying that the amino acid composition meets product specifications.

The sample is hydrolyzed using acid (and optionally enzymatic pre-treatment) to release amino acids. After derivatization to enhance detection, the amino acids are separated by HPLC or ion-exchange chromatography and quantified against known standards. Internal standards and duplicate runs provide correction for recovery and ensure method precision.

Results are reported as mg per 100 g or as a percentage of total protein. Each amino acid’s value is compared against nutritional requirements or target formulation standards. Deviations (e.g., low lysine) may indicate processing losses or protein damage and prompt further investigation.

This test quantifies andrographolides — primarily andrographolide and its analogs — in Andrographis paniculata extracts, dietary supplements, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Andrographolides are the principal bioactive diterpene lactones responsible for the immune-modulating and anti-inflammatory properties of Andrographis, and their concentration is the standard marker used to define extract potency and standardization. Results are reported in mg per serving or as a percentage of extract weight to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a methanol or ethanol-water solvent system with sonication to ensure complete recovery of andrographolide and related diterpene lactones from the botanical matrix. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 223–254 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified andrographolide reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Andrographolide content is the defining quality marker for Andrographis paniculata extracts, and accurate quantification is essential for verifying that a raw material or finished product meets its stated standardization level. HPLC-UV provides the selectivity needed to resolve andrographolide and its key analogs from the complex mixture of co-extractives present in botanical matrices, delivering reliable potency data for both incoming raw material qualification and finished product release testing.

This test quantifies total anthocyanidins — the aglycone forms of anthocyanins responsible for the characteristic deep blue-purple pigmentation and antioxidant activity of blueberry (Vaccinium corymbosum and related species) — in blueberry extracts, raw materials, and dietary supplements using UV-Visible (UV-Vis) Spectrophotometry. Anthocyanidins are widely recognized for their antioxidant, anti-inflammatory, and neuroprotective properties, and total anthocyanidin content is a key potency marker used to standardize blueberry extracts. UV-Vis provides a rapid, cost-effective method for total anthocyanidin quantification appropriate for raw material screening and quality control. Results are reported as a percentage or in milligrams per gram, typically expressed as cyanidin-3-glucoside equivalents.

A representative sample is accurately weighed and extracted using acidified aqueous methanol or ethanol (e.g., 1% HCl in methanol) to ensure complete extraction and stabilization of the pH-sensitive anthocyanidin pigments. The extract is filtered and its absorbance is measured spectrophotometrically at the characteristic anthocyanidin absorption maximum of approximately 520–535 nm against a solvent blank. Total anthocyanidin concentration is calculated using the Beer-Lambert law with the molar extinction coefficient of the reference anthocyanidin (typically cyanidin-3-glucoside), or by comparison to a multi-point calibration curve prepared from a certified reference standard. All measurements are performed in triplicate and averaged for final quantification.

Total anthocyanidin content by UV-Vis is the standard quality control method for blueberry and berry-derived extracts, providing a rapid and accessible measure of pigment potency that is well correlated with antioxidant activity. While UV-Vis does not resolve individual anthocyanidin species, it is appropriate for standardized blueberry extracts where the anthocyanin profile is well characterized and total pigment content is the primary specification parameter. This method supports efficient raw material screening and supplier qualification, with HPLC available as a complementary method when individual anthocyanidin profiling is required.

This assay quantifies apigenin, a naturally occurring flavonoid found in chamomile, parsley, and citrus plants. Using LC-MS/MS, it verifies apigenin content in supplements, teas, and botanical blends to confirm label claims and ensure consistent dosing in formulations targeting mood, inflammation, and longevity.

Samples are extracted using alcohol or aqueous solvents depending on the matrix, then analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified apigenin standards with internal standard correction and duplicate injections for precision.

Results are reported in mg per g or per serving. Values are compared to formulation targets and declared label claims to confirm active compound presence and detect variability across batches.

This test quantifies total withanolides — a class of naturally occurring C28 steroidal lactones that are the principal bioactive constituents and primary standardization markers of ashwagandha (Withania somnifera (L.) Dunal) root and root extract — using HPLC with UV detection and/or LC-MS/MS. The withanolide family includes withaferin A, withanolide A, withanolide D, withanone, and numerous related steroidal lactone glycosides (withanosides), with withaferin A and withanolide A being the most pharmacologically characterized. Total withanolide content is the standard potency specification for ashwagandha extracts in the dietary supplement industry, with commercial standardized extracts typically containing 2.5–35% total withanolides depending on the extraction process and product tier. Accurate quantification is essential for label claim substantiation, raw material qualification, and ensuring batch-to-batch consistency. Results are reported as a percentage or in milligrams per gram or per serving of total withanolides, with individual withanolide species reported where LC-MS/MS profiling is applied.

A representative sample is accurately weighed and extracted using an appropriate organic solvent system (e.g., methanol or aqueous methanol) to ensure complete extraction of the withanolide fraction. For HPLC quantification, the extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 227 nm, the characteristic absorption maximum of the withanolide α,β-unsaturated δ-lactone chromophore. For LC-MS/MS profiling and quantification, the extract is analyzed using electrospray ionization (ESI) in positive ion mode with MRM transitions selected for individual withanolide species. Quantification is performed against a multi-point external calibration curve prepared from certified withanolide reference standards (e.g., withaferin A, withanolide A). Total withanolides are reported as the sum of all individually quantified withanolide species or, for HPLC-UV methods, as withaferin A equivalents. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Withanolides' α,β-unsaturated δ-lactone chromophore provides characteristic UV absorption at 227 nm, making HPLC with UV detection a practical and widely used method for total withanolide quantification in ashwagandha extracts. LC-MS/MS provides additional compound-specific selectivity for individual withanolide species profiling, enabling discrimination between pharmacologically distinct withanolides (e.g., withaferin A vs. withanolide A) and detection of adulteration or blending with withanolide-poor ashwagandha materials. The combination of both techniques supports comprehensive potency characterization, label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay qualitatively evaluates samples for the presence of Withania somnifera using High-Performance Thin-Layer Chromatography (HPTLC). The test compares characteristic chromatographic fingerprints to authenticated reference material to confirm botanical presence.

Samples are extracted and applied to an HPTLC plate alongside reference standards. After chromatographic development and visualization under appropriate conditions, the resulting banding pattern is evaluated for correspondence with the reference profile to determine presence or absence.

Testing supports ingredient verification, supplier qualification, and quality control for botanical materials without making potency claims.

This assay quantifies both free and esterified forms of astaxanthin, a powerful antioxidant carotenoid found naturally in algae and marine sources. Testing ensures standardized potency and verifies the presence of bioactive carotenoid compounds.

Samples are extracted and analyzed under validated chromatographic conditions to distinguish and quantify free astaxanthin and its mono- and di-ester derivatives. Certified reference standards and quality controls ensure accurate and reproducible results.

Testing confirms ingredient standardization, verifies label claims, and supports product consistency across batches.

This test measures the concentration of synthetic astaxanthin, a potent antioxidant carotenoid, in raw materials, finished products, capsules, and powders using High-Performance Liquid Chromatography (HPLC). The method accurately separates geometric isomers of astaxanthin to provide precise quantification, with results reported in milligrams per gram or per serving. This ensures quality control and label compliance for products containing synthetic astaxanthin.

Samples are first extracted with a mixture of acetone and hexane to efficiently solubilize astaxanthin and remove interfering substances. The extract is then filtered and injected into an HPLC system equipped with a C18 reversed-phase column. Detection is performed using a UV-Vis detector set at 478 nm, targeting the characteristic absorbance of astaxanthin. Quantification is achieved by comparing peak areas to a calibration curve constructed from certified synthetic astaxanthin reference standards. Method accuracy is verified through duplicate injections, inclusion of quality control samples, and spike recovery tests to confirm extraction efficiency.

Results are reported in mg/g (raw materials) or mg/serving (finished products). Testing confirms standardized potency, supports quality control, and ensures batch-to-batch consistency in synthetic astaxanthin products.

This test confirms the botanical identity of Bacopa monnieri in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic bacosides A and B fingerprint of the sample is compared against a certified Bacopa monnieri reference standard to confirm species authenticity and detect substitution with other Bacopa species or unrelated botanical materials. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or ethanol-water and applied alongside a certified Bacopa monnieri reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, derivatized with anisaldehyde-sulfuric acid or natural products reagent, and the fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Bacopa monnieri is a high-value nootropic botanical where accurate species identification is essential for confirming the presence of the characteristic bacoside compounds responsible for its cognitive benefits. HPTLC identity testing provides a rapid and defensible species confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies Bacosides A and B, saponin glycosides found in Bacopa monnieri, using High-Performance Liquid Chromatography (HPLC). Results verify extract standardization and ensure compliance with product specifications.

This test measures the concentrations of Bacosides A and B, key active saponin glycosides in Bacopa monnieri extracts, using High-Performance Liquid Chromatography (HPLC). Accurate quantification of these compounds in raw materials, powders, and finished products ensures extract standardization and compliance with quality specifications. The method reports results in percentage weight per weight (% w/w) with detection limits suitable for typical supplement formulations.

Samples are prepared by solvent extraction using methanol to isolate Bacosides A and B from the matrix. The extract is filtered and injected into an HPLC system equipped with a reversed-phase C18 column. Detection is performed using UV absorbance at 205 nm, where Bacosides A and B exhibit strong signals. Quantification is achieved by comparing peak areas to a calibration curve constructed from certified reference standards of Bacosides A and B. Method precision and accuracy are verified through duplicate injections, quality control samples, and spike recovery experiments.

This test quantifies bacosides — the primary bioactive triterpenoid saponin glycosides found in Bacopa monnieri (Brahmi), including bacoside A and bacoside B and their component glycosides — in botanical extracts, raw materials, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Bacosides are the principal marker compounds used to standardize Bacopa extracts and are directly linked to the herb's documented effects on memory consolidation, cognitive performance, and neuroprotection. HPLC provides greater specificity than UV-Vis colorimetric methods by resolving individual bacoside components, offering a more detailed and accurate measure of extract potency. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and extracted using aqueous methanol or ethanol, with sonication to ensure complete dissolution of the saponin glycosides. The extract is filtered through a 0.2 µm membrane and analyzed by reversed-phase HPLC on a C18 column, with UV or Photodiode Array (PDA) detection at approximately 205–210 nm. Individual bacoside peaks are resolved chromatographically and quantified against a multi-point external calibration curve prepared from certified bacoside A or total bacoside reference standards. System suitability and quality control standards are run concurrently to confirm method accuracy and precision throughout the analytical run.

Bacopa monnieri extracts are commercially standardized to a declared bacoside content, and HPLC quantification provides a more specific and reproducible measure of individual bacoside components than colorimetric UV-Vis methods, which measure total saponin content without distinguishing between individual glycosides. This level of specificity is important for premium extract qualification, where the relative composition of individual bacosides may influence biological activity. HPLC potency testing supports label claim accuracy, supplier qualification, and cGMP compliance under 21 CFR 111.

This test quantifies baicalin — the primary bioactive flavone glucuronide found in the roots of Scutellaria baicalensis (Chinese skullcap) — in botanical extracts, raw materials, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Baicalin is the principal marker compound used to standardize Scutellaria baicalensis root extracts and is associated with the herb's well-documented antioxidant, anti-inflammatory, and neuroprotective activity. Accurate quantification confirms that the extract meets its declared potency and that the correct botanical species is present, which is particularly important given the known risk of adulteration with other Scutellaria species or unrelated botanicals. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and extracted using aqueous methanol or ethanol, with sonication or reflux to ensure complete extraction of the flavonoid fraction from the botanical matrix. The extract is filtered through a 0.2 µm membrane and analyzed by reversed-phase HPLC on a C18 column, with UV or Photodiode Array (PDA) detection at approximately 277 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified baicalin reference standard. Peak identity is confirmed by retention time and UV spectral comparison to the reference standard, and system suitability and quality control standards are run concurrently to confirm method accuracy and precision throughout the analytical run.

Scutellaria baicalensis extracts are commercially standardized on the basis of baicalin content, making accurate HPLC quantification the primary means of verifying extract potency and authenticity. The genus Scutellaria has also been associated with adulteration using Teucrium species (germander), which contain hepatotoxic neo-clerodane diterpenes and lack baicalin — making baicalin quantification not only a potency check but also an indirect indicator of species authenticity. HPLC provides the chromatographic resolution needed to specifically quantify baicalin and distinguish it from related flavonoids such as baicalein and wogonoside in complex botanical matrices, supporting raw material qualification and label claim compliance under 21 CFR 111.

This assay quantifies the three key branched-chain amino acids (BCAAs)—leucine, isoleucine, and valine—using LC-MS/MS. Commonly found in protein powders, amino blends, and pre-workouts, this test verifies BCAA content and ratios to ensure compliance with label claims and prevent misformulation or ingredient dilution.

Samples are hydrolyzed or extracted depending on the matrix, then filtered and analyzed by LC-MS/MS. Detection is based on compound-specific mass transitions. Quantification is performed using high-purity amino acid standards, with internal standard correction and duplicate runs to ensure accuracy and reproducibility.

Results are reported in mg/g or mg per serving. Values are compared to declared label claims and standard 2:1:1 or other specified ratios. Testing confirms proper dosing and supports claims related to muscle recovery, endurance, and anabolic performance in sports nutrition products.

This test confirms the botanical identity of beet root (Beta vulgaris) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic betalain pigment fingerprint — including betanin and vulgaxanthin — of the sample is compared against a certified Beta vulgaris reference standard to confirm species authenticity and detect substitution with other red-pigmented botanicals or synthetic colorants. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using a methanol-water solvent system and applied alongside a certified beet root reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system and examined under visible light and UV (366 nm) to visualize the characteristic betalain bands. The resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Beet root powder is susceptible to adulteration with synthetic red dyes or other red-pigmented botanical powders that visually resemble the authentic material. HPTLC identity testing provides a rapid and defensible confirmation of botanical species, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies berberine, an isoquinoline alkaloid commonly sourced from plants like Berberis and used in supplements for metabolic and cardiovascular support. Using HPLC, it measures berberine content in capsules, tablets, and botanical extracts to verify potency and ensure formulation accuracy.

Samples are extracted using alcohol- or acidified aqueous solvents and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified berberine standards, with internal standard correction and duplicate runs to ensure precision.

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm consistent dosing and detect degradation or ingredient substitution.

This test quantifies berberine hydrochloride — an isoquinoline alkaloid and the primary bioactive constituent of botanical sources such as barberry (Berberis vulgaris), goldenseal (Hydrastis canadensis), and Oregon grape (Mahonia aquifolium) — in raw materials, extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Berberine HCl is one of the most widely studied botanical actives, with a substantial body of clinical evidence supporting its role in glycemic control, lipid metabolism, and cardiovascular health. Accurate potency verification is essential for label claim substantiation and for confirming that the declared amount of this high-value alkaloid is present, particularly given the prevalence of adulteration and potency variability in berberine-containing raw materials. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate solvent, typically dilute hydrochloric acid in methanol or aqueous methanol, to ensure complete extraction of berberine and its salt form. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 345 nm, the characteristic absorption maximum of berberine's quaternary ammonium chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified berberine hydrochloride reference standard. Where applicable, related alkaloids such as palmatine, coptisine, and jatrorrhizine may be monitored simultaneously to provide a more complete alkaloid profile. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Berberine's strong UV absorption at 345 nm makes HPLC with UV detection a highly sensitive and specific method for its quantification without the need for derivatization. The chromatographic separation resolves berberine from structurally related isoquinoline alkaloids that co-occur in berberine-containing botanicals, ensuring that potency results reflect the berberine HCl content specifically rather than total alkaloid content. This level of specificity is important for label claim accuracy and for detecting adulteration or dilution with lower-cost alkaloid-containing materials. The method supports raw material qualification, finished product release testing, and cGMP compliance under 21 CFR 111.

This assay quantifies beta-alanine, a non-essential amino acid widely used in sports supplements to buffer lactic acid and delay fatigue. Using LC-MS/MS, it confirms the presence and potency of beta-alanine in pre-workouts, amino blends, and performance formulas to support label accuracy and product efficacy.

Samples are extracted in aqueous solution, filtered, and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified beta-alanine standards with internal standard correction and duplicate injections to ensure precision.

Results are reported in mg per g or per serving. Values are compared with formulation targets and declared label claims. Testing ensures that efficacious doses are delivered consistently across batches and helps guard against under-dosing or mislabeling.

This test quantifies beta-caryophyllene, a key terpene known for its aromatic and therapeutic properties, using Gas Chromatography–Mass Spectrometry (GC-MS). It is applicable to essential oils, botanical extracts, and finished products where terpene profiling is critical for quality control. The method provides sensitive detection with quantification limits typically in the low microgram per gram range.

Samples are prepared by diluting the botanical extract or essential oil in an appropriate organic solvent such as hexane. The prepared solution is injected into the GC-MS system equipped with a non-polar capillary column. Beta-caryophyllene is separated based on retention time and identified by matching its mass spectral fragmentation pattern to certified reference standards. Quantification is performed using an external calibration curve constructed from known concentrations of beta-caryophyllene standards. Quality control includes duplicate injections, the use of internal standards to correct for variability, and spike recovery tests to confirm accuracy.

Results are reported in % w/w (raw materials) or mg/g (finished products). Testing confirms product standardization, supports quality control, and ensures compliance with labeling and purity specifications.

This assay quantifies beta-glucan content using the enzymatic Megazyme method, which is AOAC- and AACC-approved for measuring beta-glucans in cereal grains, mushrooms, and dietary supplements. It is commonly used for standardizing oat and barley extracts as well as immune-support formulations.

Samples are enzymatically digested using lichenase and β-glucosidase to release glucose from beta-glucan polymers. The liberated glucose is then measured spectrophotometrically using glucose oxidase/peroxidase reagents.

Results are reported in % w/w or mg/g of beta-glucan. Values are compared to standardization targets and label claims to ensure active fiber content and formulation consistency.

This test quantifies beta-glucan — a soluble polysaccharide found in oats, barley, and fungal sources such as yeast and medicinal mushrooms — in raw materials, extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Beta-glucan is one of the most extensively studied functional ingredients, with FDA-authorized health claims for oat and barley beta-glucan's role in reducing the risk of heart disease. Accurate potency verification is essential for substantiating label claims, confirming the source-appropriate beta-glucan structure (cereal vs. fungal), and ensuring that the declared amount of this functional fiber is present in the finished product. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and subjected to a controlled enzymatic hydrolysis protocol to selectively degrade beta-glucan into its constituent glucose units while minimizing interference from starch and other polysaccharides. Starch is first removed by treatment with amyloglucosidase and invertase, followed by specific hydrolysis of the beta-glucan fraction using lichenase (endo-1,3:1,4-β-glucanase). The resulting oligosaccharide hydrolysate — primarily tri- and tetrasaccharides characteristic of (1→3),(1→4)-β-D-glucan — is analyzed by HPLC on a suitable carbohydrate column with refractive index (RI) or evaporative light scattering detection (ELSD). Quantification is performed against a multi-point external calibration curve prepared from a certified beta-glucan reference standard. Quality control samples are run concurrently to confirm method accuracy and precision.

Beta-glucan is a structurally complex polysaccharide whose potency cannot be accurately measured by simple gravimetric or total fiber methods, which do not distinguish between beta-glucan and other dietary fiber components. HPLC following selective enzymatic hydrolysis provides a specific and reproducible measure of true beta-glucan content, distinguishing it from starch, cellulose, and other polysaccharides in complex cereal and fungal matrices. This method is aligned with AOAC-validated approaches for beta-glucan quantification and supports FDA health claim compliance, label claim substantiation, and raw material qualification under 21 CFR 111.

This test quantifies betaine (trimethylglycine, TMG) — a naturally occurring trimethyl derivative of the amino acid glycine found in foods such as beets (Beta vulgaris), wheat germ, and spinach — in raw materials and dietary supplements using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Betaine functions as an osmolyte and methyl donor in one-carbon metabolism, supporting homocysteine remethylation, liver fat metabolism, and cellular hydration. It is widely used in dietary supplements for cardiovascular health, liver support, and sports performance applications. Accurate potency verification by LC-MS/MS is essential for specifically quantifying betaine and distinguishing it from structurally related compounds — including betaine aldehyde, choline, and carnitine — that may co-occur in complex botanical and food-derived matrices. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate aqueous diluent. An isotopically labeled internal standard (e.g., betaine-d9 or ¹³C-labeled betaine) is added prior to sample preparation to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase or HILIC LC-MS/MS using electrospray ionization (ESI) in positive ion mode, with multiple reaction monitoring (MRM) transitions selected for the characteristic precursor and product ions of betaine (m/z 118 → 58 and 118 → 59 are commonly used transitions). Quantification is performed against a multi-point external calibration curve prepared from a certified betaine reference standard. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Betaine is a small, highly polar, permanently charged quaternary ammonium compound with no significant UV chromophore, making it poorly suited to standard reversed-phase HPLC with UV detection. LC-MS/MS with MRM detection provides the compound-specific selectivity and sensitivity required to accurately quantify betaine in complex food and supplement matrices, distinguishing it from isobaric and structurally related compounds such as choline, carnitine, and proline betaine that may co-occur and interfere with less specific analytical methods. The use of an isotopically labeled internal standard further ensures accurate quantification by correcting for matrix-dependent ionization effects. This method supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This test quantifies betaine nitrate — a salt combining trimethylglycine (betaine) and nitrate, used in sports nutrition formulations for its combined vasodilatory and performance-enhancing properties — in raw materials and dietary supplements using titration. Accurate quantification of betaine nitrate is essential for label claim verification and for confirming that the declared dose of this dual-function ingredient is present in the finished product. Titration provides a reliable, direct measure of the betaine nitrate content in high-purity raw material testing. Results are reported as a percentage purity or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in a suitable solvent. Depending on the validated protocol, either the nitrate component or the betaine component is targeted for titration. For nitrate quantification, a potentiometric or argentometric titration approach may be applied. For betaine quantification, a non-aqueous perchloric acid titration in glacial acetic acid may be used, exploiting the basic character of the trimethylglycine moiety. The endpoint is determined potentiometrically or by indicator color change, and the betaine nitrate content is calculated from the volume and molarity of titrant consumed relative to the sample weight. Reference standard checks and blank titrations are performed concurrently to confirm method accuracy.

Betaine nitrate is a relatively novel, premium sports nutrition ingredient where both the betaine and nitrate components contribute to its performance benefits, making accurate potency verification important for both label claim compliance and product efficacy. Titration provides a straightforward and cost-effective method for confirming the purity of betaine nitrate raw materials prior to use in formulation, supporting incoming material qualification and cGMP compliance under 21 CFR 111. For finished product testing in complex multi-ingredient matrices, complementary methods such as HPLC or IC may be required to provide adequate specificity.

This assay measures beta-sitosterol, the primary plant sterol found in seeds, nuts, and botanical oils. LC-MS/MS provides high sensitivity and specificity for distinguishing beta-sitosterol from other sterols in complex lipid matrices.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Beta-sitosterol is detected using multiple reaction monitoring (MRM) and quantified against certified reference standards. Internal calibration and quality control checks ensure consistent accuracy and reproducibility.

Testing verifies standardized potency, supports label claims, and confirms purity in plant-sterol–containing formulations.

This test quantifies piperine — the primary bioactive alkaloid of black pepper (Piper nigrum L.) and the active constituent of the proprietary standardized extract BioPerine® (Sabinsa Corporation) — in raw materials and dietary supplements using High-Performance Liquid Chromatography (HPLC). BioPerine® is standardized to a minimum of 95% piperine and is widely incorporated into dietary supplement formulations as a bioavailability-enhancing ingredient, with a body of clinical research supporting its role in increasing the absorption of a broad range of nutrients and pharmaceuticals through inhibition of intestinal and hepatic metabolism. Accurate piperine quantification is essential for verifying that BioPerine®-containing products meet the standardized potency specification and deliver the intended bioavailability-enhancing dose. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate solvent, typically methanol or aqueous methanol, to ensure complete extraction of piperine. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 343 nm, the characteristic absorption maximum of piperine's conjugated diene chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified piperine reference standard. Where applicable, related piperamide alkaloids (e.g., piperyline, piperettine) may be monitored to assess extract purity and confirm the characteristic alkaloid profile of P. nigrum. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Piperine's strong UV absorption at 343 nm makes HPLC with UV detection a highly sensitive and specific method for its quantification, providing the precision required to verify compliance with BioPerine®'s standardized ≥95% piperine specification. Chromatographic separation resolves piperine from structurally related piperamide alkaloids and matrix components, ensuring that potency results reflect piperine content specifically. This is particularly important for finished supplement matrices where BioPerine® is combined with other botanical extracts that may contain UV-absorbing compounds. The method supports raw material qualification, finished product release testing, and cGMP compliance under 21 CFR 111.

This test confirms the identity of bison (Bison bison) liver in raw materials, freeze-dried liver powders, and dietary supplements using Fourier Transform Infrared Spectroscopy (FTIR). Bison liver is a whole food ingredient used in ancestral nutrition and organ meat supplement formulations, valued for its exceptional density of bioavailable nutrients including heme iron, vitamin A (retinol), B vitamins (particularly B12 and folate), copper, and coenzyme Q10. FTIR identity testing generates a characteristic mid-infrared absorption spectrum reflecting the composite molecular composition — including proteins, lipids, carbohydrates, and nucleic acids — of the liver matrix. This spectral fingerprint is compared against an authenticated bison liver reference spectrum to confirm material identity and detect potential substitution with liver from other species (e.g., bovine, porcine, or chicken) or with non-liver organ tissues.

A representative sample is accurately weighed and prepared for FTIR analysis using an appropriate sample presentation technique. For dried or powdered materials, attenuated total reflectance (ATR-FTIR) is the preferred approach, in which the sample is placed directly onto the ATR crystal (typically diamond or zinc selenide) and compressed with a pressure applicator to ensure adequate optical contact. The mid-infrared spectrum is collected over the range of 4000–400 cm⁻¹ at a defined spectral resolution (typically 4 cm⁻¹) with an appropriate number of co-added scans for signal averaging. The resulting spectrum is baseline-corrected and compared against an authenticated bison liver reference spectrum library using spectral correlation or chemometric matching algorithms. Identity is confirmed when the sample spectrum meets a defined similarity threshold relative to the reference spectrum, and is differentiated from spectra of other species' liver materials and non-liver tissues.

FTIR spectroscopy provides a rapid, non-destructive, and reagent-free method for identity confirmation of complex biological matrices such as organ meat powders, generating a holistic molecular fingerprint that reflects the composite composition of the material. For organ meat ingredients such as bison liver, where species authentication and tissue-type confirmation are both required, FTIR provides a practical first-line identity screening tool that can distinguish bison liver from other species' liver materials and from non-liver organ tissues based on differences in protein secondary structure, lipid composition, and overall matrix chemistry. This method supports raw material qualification, supplier verification, and cGMP compliance under 21 CFR 111.

This test confirms the identity of bitter melon (Momordica charantia L.) in raw materials, fruit powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Momordica charantia, commonly known as bitter melon, bitter gourd, or karela, is a tropical vine used extensively in traditional Ayurvedic, Chinese, and Caribbean medicine for the management of blood glucose levels and metabolic health. Its characteristic phytochemical profile includes cucurbitane-type triterpenoids (notably momordicosides and charantin), steroidal saponins, alkaloids (vicine), and polypeptide-p. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated M. charantia reference standard to confirm species identity and detect potential adulteration, substitution with other Momordica species, or blending with unrelated plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic secondary metabolite profile of M. charantia, including triterpenoids and saponins. The extract is applied alongside a certified bitter melon reference standard and, where applicable, potential adulterant extracts, onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve the characteristic marker compounds of M. charantia. After development, the plate is derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid or vanillin-sulfuric acid) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Bitter melon raw materials are subject to adulteration and species substitution, including blending with other Momordica species or unrelated cucurbit plant materials, particularly given the variability in raw material quality across global supply chains. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that is more discriminating than single-marker assays, enabling detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test confirms the identity of black currant (Ribes nigrum L.) in raw materials, berry powders, juice powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Ribes nigrum, commonly known as black currant, is a small berry native to northern Europe and Asia, widely used in dietary supplements for its exceptionally high anthocyanin content and potent antioxidant capacity. Its characteristic phytochemical profile includes anthocyanins (primarily delphinidin-3-O-glucoside, delphinidin-3-O-rutinoside, cyanidin-3-O-glucoside, and cyanidin-3-O-rutinoside), flavonols (quercetin, myricetin, and kaempferol glycosides), hydroxycinnamic acids (chlorogenic acid, neochlorogenic acid), and vitamin C. HPTLC identity testing generates a characteristic chromatographic fingerprint — anchored by the distinctive anthocyanin and flavonol profile — that is compared against an authenticated R. nigrum reference standard to confirm species identity and detect potential adulteration, substitution with other Ribes species or berry materials, or blending with inferior fruit powders.

A representative sample is accurately weighed and extracted using an appropriate acidified solvent system (e.g., acidified methanol or aqueous ethanol with 0.1% hydrochloric acid) to ensure complete extraction and stabilization of the anthocyanin fraction alongside the flavonol and phenolic acid constituents. The extract is applied alongside a certified R. nigrum reference standard and, where applicable, potential adulterant extracts (e.g., other Ribes species, blueberry, elderberry), onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve the characteristic anthocyanin and flavonol constituents of R. nigrum. After development, the plate is evaluated under white light — where anthocyanins are visible as characteristic pink-purple bands — and under UV light at 254 nm and 366 nm. Derivatization with Natural Products Reagent A (NP/PEG) may be applied for enhanced visualization of flavonol glycosides under UV 366 nm. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Black currant raw materials are subject to adulteration and species substitution, including replacement with other Ribes species (e.g., red currant, R. rubrum; blackcurrant hybrids), other dark berry materials (e.g., elderberry, blueberry, chokeberry), or dilution with added colorants or inferior fruit powders. The characteristic anthocyanin profile of R. nigrum — dominated by delphinidin and cyanidin glycosides in a distinctive ratio — provides a highly discriminating fingerprint that distinguishes authentic black currant from other berry materials with different anthocyanin compositions. HPTLC fingerprinting provides a holistic, multi-compound identity confirmation that is more discriminating than single-marker anthocyanin content assays, enabling detection of substitution or adulteration that would not be apparent from total polyphenol or ORAC testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This assay confirms the identity of Black Ginger (Kaempferia parviflora) using High-Performance Thin-Layer Chromatography (HPTLC). The method produces a unique fingerprint of flavonoid compounds (such as polymethoxyflavones) that distinguishes Black Ginger from adulterants and other ginger family botanicals.

Samples are extracted with alcohol-based solvents and applied to a silica gel HPTLC plate alongside authenticated Black Ginger reference material. Plates are developed in a suitable mobile phase and visualized under UV light and/or after derivatization. The resulting chromatographic fingerprint is compared against the reference standard to confirm authenticity.

Results are reported as Pass/Fail or Match/No Match to the reference standard. HPTLC is a pharmacopeial method widely accepted for botanical identity testing, ensuring that Black Ginger extracts are genuine and not substituted with other ginger species.

This test confirms the identity of black pepper (Piper nigrum L.) in raw materials, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Piper nigrum is one of the most extensively used spices and botanical ingredients globally, and its primary bioactive alkaloid, piperine, is widely incorporated into dietary supplement formulations as a bioavailability enhancer (commonly marketed under proprietary names such as BioPerine®). HPTLC identity testing generates a characteristic chromatographic fingerprint — anchored by the prominent piperine band alongside other characteristic alkaloids and amides — that is compared against an authenticated P. nigrum reference standard to confirm species identity and detect potential adulteration, substitution with other Piper species (e.g., P. longum, P. retrofractum), or blending with inferior materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or dichloromethane) to capture the characteristic alkaloid and amide profile of P. nigrum, including piperine and related piperamides. The extract is applied alongside a certified black pepper reference standard and, where applicable, potential adulterant or related Piper species extracts, onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve piperine and the characteristic secondary metabolites of P. nigrum. After development, the plate is evaluated under UV light at 254 nm and 366 nm, and may be further derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid) for visualization under white light. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Black pepper and closely related Piper species share similar morphological characteristics, making botanical identity confirmation by HPTLC fingerprinting essential for distinguishing authentic P. nigrum from potential adulterants or substitutes. HPTLC provides a holistic, multi-compound chromatographic identity confirmation that is more discriminating than single-marker piperine assays alone, enabling detection of species substitution or adulteration that would not be apparent from potency testing. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This assay quantifies elemental boron in supplements, raw materials, or functional foods using Inductively Coupled Plasma Mass Spectrometry (ICP-MS). It is commonly used to verify boron content in bone health and trace mineral formulations or ensure compliance with formulation specifications.

Samples are digested using acid-based microwave or wet digestion protocols and analyzed by ICP-MS. Quantification is performed using certified boron standards with internal standard correction and quality control samples to ensure accurate and reproducible results.

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm mineral content, detect overages, and ensure dosing consistency.

This test confirms the identity of boswellia (Boswellia serrata Roxb. ex Colebr., and related pharmacopeial species including B. sacra, B. carterii, and B. frereana) in raw materials, gum resin powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Boswellia, commonly known as Indian frankincense, is one of the most extensively researched botanical anti-inflammatory ingredients, with its bioactivity attributed primarily to boswellic acids — a family of pentacyclic triterpenic acids including β-boswellic acid, acetyl-β-boswellic acid (ABA), 11-keto-β-boswellic acid (KBA), and acetyl-11-keto-β-boswellic acid (AKBA). HPTLC identity testing generates a characteristic chromatographic fingerprint anchored by the boswellic acid profile that is compared against an authenticated Boswellia reference standard to confirm species identity and detect potential adulteration, substitution with other resinous botanicals, or blending with inferior materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol, ethanol, or dichloromethane) to capture the characteristic triterpenoid acid profile of Boswellia spp. The extract is applied alongside a certified boswellia reference standard and, where applicable, potential adulterant extracts, onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated non-polar to moderately polar solvent system optimized to resolve the characteristic boswellic acid constituents of Boswellia spp. After development, the plate is derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid or vanillin-sulfuric acid) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting fingerprint pattern — characterized by the distinctive band pattern of boswellic acids — is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Boswellia raw materials are subject to adulteration and species substitution, including blending with other Boswellia species, non-pharmacopeial resin sources, or synthetic diluents, particularly given the premium pricing of high-AKBA standardized extracts. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that is more discriminating than single-marker boswellic acid assays, enabling detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test identifies and quantifies boswellic acids, the active compounds in Boswellia extracts, which are important for product efficacy and quality control. The method uses liquid chromatography-tandem mass spectrometry (LC-MS/MS) to provide sensitive and specific detection in raw materials, powders, and finished products. Results are reported in mg/g with detection limits suitable for trace-level analysis.

Samples are prepared by extracting with methanol under controlled temperature and agitation to ensure efficient recovery of boswellic acids. The extracts are analyzed using LC-MS/MS in multiple reaction monitoring (MRM) mode, targeting specific mass transitions for each boswellic acid. Quantification is achieved through calibration curves constructed with certified boswellic acid reference standards and corrected using an internal standard. Method accuracy and precision are verified by duplicate injections and spike recovery tests.

Results are reported via Present or Not Present. Values are compared to standardization targets and label claims to verify active content and detect underformulation or degradation.

This assay measures bovine epidermal growth factor (EGF), a peptide growth factor present in bovine colostrum and dairy-derived ingredients. Analysis may be performed using ELISA for immuno-specific quantification or LC-MS/MS for confirmatory peptide-level measurement.

Samples are prepared and analyzed using validated ELISA and/or LC-MS/MS workflows.

ELISA employs bovine-specific antibodies to quantify EGF relative to calibrated standards.

LC-MS/MS provides orthogonal confirmation by detecting EGF-specific peptide fragments following protein digestion using multiple reaction monitoring (MRM).

Quality controls and reference materials ensure accuracy and reproducibility.

Testing verifies bovine EGF content, supports label claims, and ensures batch-to-batch consistency in colostrum and bioactive protein products.

This assay measures bovine insulin-like growth factor-1 (IGF-1), a peptide growth factor present in bovine colostrum and dairy-derived ingredients. Analysis may be performed using ELISA for targeted immunoquantification or LC-MS/MS for confirmatory peptide-level measurement.

Samples are prepared and analyzed using validated ELISA and/or LC-MS/MS workflows.

ELISA uses bovine-specific antibodies to quantify IGF-1 against calibrated standards.

LC-MS/MS provides orthogonal confirmation by detecting IGF-1–specific peptide fragments following protein digestion.

Quality controls and reference materials ensure accuracy and reproducibility.

Testing verifies bovine IGF-1 content, supports label claims, and ensures batch-to-batch consistency.

This assay measures bovine insulin-like growth factor-2 (IGF-2), a peptide growth factor present in bovine colostrum and dairy-derived ingredients. LC-MS/MS provides highly specific and sensitive quantification through peptide-level detection.

Samples are prepared and analyzed under validated LC-MS/MS chromatographic conditions. Following protein digestion, IGF-2–specific peptide fragments are detected using multiple reaction monitoring (MRM) and quantified against certified reference standards. Internal calibration and quality controls ensure accuracy and reproducibility.

Testing verifies bovine IGF-2 content, supports label claims, and ensures batch-to-batch consistency in colostrum and bioactive protein products.

This assay measures bovine immunoglobulin M (IgM), an antibody class present in bovine colostrum and dairy-derived ingredients. Analysis may be performed using ELISA for targeted immunoreactivity or LC-MS/MS for confirmatory protein quantification, depending on application needs.

Samples are prepared and analyzed using validated ELISA and/or LC-MS/MS methods.

ELISA uses bovine-specific antibodies to selectively detect IgM and quantify concentration via comparison to calibrated standards.

LC-MS/MS provides orthogonal confirmation by detecting IgM-specific peptide fragments following protein digestion.

Quality controls and reference materials ensure accuracy and reproducibility.

Testing verifies bovine IgM content, supports label claims, and ensures batch-to-batch consistency in colostrum and immune-focused products.

This test confirms the identity of bovine liver (Bos taurus) in raw materials, freeze-dried liver powders, and dietary supplements using Fourier Transform Infrared Spectroscopy (FTIR). Bovine liver is a whole food ingredient widely used in ancestral nutrition and organ meat supplement formulations, valued for its exceptional density of bioavailable nutrients including heme iron, vitamin A (retinol), vitamin B12, folate, riboflavin, copper, and coenzyme Q10. FTIR identity testing generates a characteristic mid-infrared absorption spectrum reflecting the composite molecular composition — including proteins, lipids, carbohydrates, and nucleic acids — of the liver matrix. This spectral fingerprint is compared against an authenticated bovine liver reference spectrum to confirm material identity and detect potential substitution with liver from other species (e.g., porcine, ovine, chicken) or with non-liver organ tissues (e.g., kidney, heart, spleen), which may be visually indistinguishable in dried and powdered form.

A representative sample is accurately weighed and prepared for FTIR analysis using attenuated total reflectance (ATR-FTIR), in which the dried powder sample is placed directly onto the ATR crystal (typically diamond or zinc selenide) and compressed with a pressure applicator to ensure adequate optical contact. The mid-infrared spectrum is collected over the range of 4000–400 cm⁻¹ at a defined spectral resolution (typically 4 cm⁻¹) with an appropriate number of co-added scans for signal averaging. The resulting spectrum is baseline-corrected and compared against an authenticated bovine liver reference spectrum using spectral correlation or chemometric library matching algorithms. Key diagnostic absorption regions — including the Amide I (~1650 cm⁻¹) and Amide II (~1540 cm⁻¹) bands reflecting protein secondary structure, the lipid C-H stretching region (~2850–2960 cm⁻¹), and the carbohydrate fingerprint region (~900–1200 cm⁻¹) — are evaluated for conformity with the reference. Identity is confirmed when the sample spectrum meets a defined similarity threshold relative to the certified bovine liver reference spectrum and is differentiated from spectra of other species' liver materials and non-liver organ tissues.

FTIR spectroscopy provides a rapid, non-destructive, and reagent-free method for identity confirmation of complex biological matrices such as organ meat powders, generating a holistic molecular fingerprint that reflects the composite protein, lipid, and carbohydrate composition of the material. For organ meat ingredients such as bovine liver, where both species authentication and tissue-type confirmation are required, FTIR provides a practical first-line identity screening tool that can distinguish bovine liver from other species' liver materials and from non-liver organ tissues based on characteristic differences in protein secondary structure, lipid composition, and overall matrix chemistry. This method supports raw material qualification, supplier verification, and cGMP compliance under 21 CFR 111.

This assay measures bovine transforming growth factor beta-1 (TGF-β1), a bioactive cytokine naturally present in bovine colostrum and dairy-derived ingredients. Analysis may be performed using ELISA for bioactive protein quantification or LC-MS/MS for confirmatory peptide-based measurement.

Samples are prepared and analyzed using validated ELISA and/or LC-MS/MS methods.

ELISA employs bovine-specific antibodies to quantify TGF-β1 concentration relative to calibrated standards.

LC-MS/MS provides orthogonal confirmation by detecting TGF-β1–specific peptide fragments following protein digestion.

Quality controls and reference materials ensure accuracy and reproducibility.

Testing verifies bovine TGF-β1 content, supports label claims, and ensures batch-to-batch consistency in colostrum and immune-focused products.

This test confirms the identity of bovine thyroid (Bos taurus) glandular material in raw materials, freeze-dried thyroid powders, and dietary supplements using Fourier Transform Infrared Spectroscopy (FTIR). Bovine thyroid is a whole food glandular ingredient used in ancestral nutrition and glandular therapy formulations, valued for its naturally occurring thyroid-associated proteins, peptides, and cofactors. FTIR identity testing generates a characteristic mid-infrared absorption spectrum reflecting the composite molecular composition — including proteins, lipids, carbohydrates, and nucleic acids — of the thyroid glandular matrix. This spectral fingerprint is compared against an authenticated bovine thyroid reference spectrum to confirm material identity and detect potential substitution with glandular material from other species (e.g., porcine, ovine) or with non-thyroid tissues (e.g., adrenal, liver, or other organ powders), which may be visually indistinguishable in dried and powdered form.

A representative sample is accurately weighed and prepared for FTIR analysis using attenuated total reflectance (ATR-FTIR), in which the dried powder sample is placed directly onto the ATR crystal (typically diamond or zinc selenide) and compressed with a pressure applicator to ensure adequate optical contact. The mid-infrared spectrum is collected over the range of 4000–400 cm⁻¹ at a defined spectral resolution (typically 4 cm⁻¹) with an appropriate number of co-added scans for signal averaging. The resulting spectrum is baseline-corrected and compared against an authenticated bovine thyroid reference spectrum using spectral correlation or chemometric library matching algorithms. Key diagnostic absorption regions — including the Amide I (~1650 cm⁻¹) and Amide II (~1540 cm⁻¹) bands reflecting the protein secondary structure composition, the lipid C-H stretching region (~2850–2960 cm⁻¹), and the carbohydrate fingerprint region (~900–1200 cm⁻¹) — are evaluated for conformity with the reference. Identity is confirmed when the sample spectrum meets a defined similarity threshold relative to the certified bovine thyroid reference spectrum and is differentiated from spectra of other species' thyroid materials and non-thyroid organ tissues.

FTIR spectroscopy provides a rapid, non-destructive, and reagent-free method for identity confirmation of complex biological matrices such as glandular organ powders, generating a holistic molecular fingerprint that reflects the composite protein, lipid, and carbohydrate composition of the material. For glandular ingredients such as bovine thyroid, where both species authentication and tissue-type confirmation are required, FTIR provides a practical first-line identity screening tool that can distinguish bovine thyroid from other species' thyroid materials and from non-thyroid organ tissues based on differences in protein secondary structure, lipid composition, and overall matrix chemistry. This method supports raw material qualification, supplier verification, and cGMP compliance under 21 CFR 111.

This test detects and quantifies bovine-derived Type II collagen in dietary supplements, raw materials, and finished products using an Enzyme-Linked Immunosorbent Assay (ELISA). Type II collagen is the predominant collagen found in articular cartilage and is widely used in joint health formulations for its role in supporting cartilage structure, reducing joint discomfort, and promoting mobility. ELISA provides highly specific immunological detection of Type II collagen, distinguishing it from other collagen types (e.g., Type I, Type III) and confirming both the species origin and collagen type present in the sample. Results are reported in micrograms per milliliter (µg/mL) or milligrams per serving to support label claim verification and cGMP compliance.

A representative sample is extracted in a suitable buffer to solubilize the collagen, with any necessary pre-treatment steps such as pepsin digestion applied to improve extraction efficiency of native or partially hydrolyzed collagen. The extract is applied to a microplate pre-coated with an antibody specific to bovine Type II collagen. After incubation and washing steps to remove unbound material, a detection antibody conjugated to an enzyme (typically horseradish peroxidase) is added, followed by a substrate solution that produces a colorimetric signal proportional to the amount of Type II collagen present. Absorbance is measured at the appropriate wavelength (typically 450 nm), and sample concentrations are calculated from a multi-point standard curve prepared from a certified bovine Type II collagen reference standard. Positive and negative controls are run with each plate to confirm assay validity.

Type II collagen is a premium ingredient that is frequently mislabeled or substituted with lower-cost Type I collagen hydrolysates, which have a different mechanism of action and are not equivalent for joint health applications. ELISA provides the immunological specificity needed to distinguish Type II collagen from other collagen types and confirm the bovine species origin — information that is critical for label claim accuracy, allergen disclosure (bovine-derived), and consumer transparency. This test supports both raw material qualification and finished product release in compliance with 21 CFR 111 cGMP requirements.

This test identifies and quantifies BPC-157 (Body Protection Compound-157), a synthetic pentadecapeptide composed of 15 amino acids, in research formulations and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). BPC-157 is used in research contexts for its studied effects on musculoskeletal repair, gut health, and angiogenesis. LC-MS/MS provides the molecular specificity required to confirm peptide identity and accurately measure concentration, distinguishing BPC-157 from degradation products, related peptide impurities, and other co-formulated compounds. Results are reported in mg per vial, mg per mL, or mg per gram as applicable.

A representative sample is dissolved or diluted in an aqueous solvent system (typically 0.1% formic acid in water/acetonitrile) and filtered prior to injection. The solution is injected onto a reversed-phase C18 HPLC column coupled to a triple quadrupole mass spectrometer operating in positive ionization Multiple Reaction Monitoring (MRM) mode. Precursor-to-product ion transitions specific to BPC-157 are monitored for both quantification and identity confirmation. Quantification is performed against a multi-point calibration curve prepared from a certified BPC-157 reference standard, with a stable isotope-labeled or structurally analogous internal standard used to correct for matrix effects and recovery variability.

BPC-157 is a synthetic peptide that requires a highly specific analytical method to confirm both identity and potency, as its molecular structure cannot be distinguished from impurities or degradation products by UV-based methods alone. LC-MS/MS provides the mass accuracy and selectivity needed to unambiguously confirm the peptide sequence and quantify it at the concentration levels relevant to research formulations, supporting product integrity and accurate labeling.

This test measures the proteolytic (protein-digesting) activity of bromelain — a mixture of cysteine proteases derived from the stem and fruit of pineapple (Ananas comosus (L.) Merr.) — in raw materials and dietary supplements using the Food Chemicals Codex (FCC) standardized activity assay. Bromelain is widely used in dietary supplements for its digestive enzyme activity, anti-inflammatory properties, and support for muscle recovery and joint health. Enzyme activity is expressed in Bromelain Casein Units (BCU) or Gelatin Digesting Units (GDU), where one GDU is defined as the amount of enzyme that digests a defined quantity of gelatin substrate per unit time under specified assay conditions. Accurate activity verification is essential for confirming that bromelain ingredient potency meets label claim specifications and for ensuring consistent functional performance across batches. Results are reported in BCU or GDU per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate cold buffer to prepare a working enzyme solution. The bromelain activity assay is performed according to the FCC monograph procedure using a casein or gelatin substrate: a defined volume of the enzyme solution is incubated with the substrate at a controlled pH (typically pH 6.0–7.0) and temperature (37°C) for a precisely timed reaction period. For the BCU assay, the reaction is terminated by addition of trichloroacetic acid (TCA) to precipitate undigested protein, and the absorbance of the TCA-soluble hydrolysate — reflecting the concentration of liberated tyrosine and tyrosine-equivalent peptides — is measured spectrophotometrically at 275 nm. Enzyme activity is calculated from the absorbance reading using the FCC-defined unit calculation, referenced against a tyrosine standard curve. Reagent blanks and substrate controls are run concurrently to correct for non-enzymatic hydrolysis and background absorbance.

The FCC bromelain activity assay is the standardized, industry-recognized method for bromelain potency measurement, providing a reproducible and internationally accepted unit of enzyme activity (BCU or GDU) that enables direct comparison of potency across different bromelain sources, suppliers, and batches. FCC-defined activity units are the standard for enzyme ingredient labeling and quality specifications in the dietary supplement industry, ensuring that label claims accurately reflect functional enzymatic potency rather than total protein mass. This approach is more meaningful for quality control of enzyme ingredients than protein content assays alone, as enzyme activity can be significantly affected by processing conditions, storage, and formulation. The method supports label claim substantiation and cGMP compliance under 21 CFR 111.

This assay quantifies bromelain activity using the USP titration method, reported in Gelatin Digesting Units (GDU). The test measures the proteolytic activity of bromelain based on its ability to hydrolyze protein substrates under standardized conditions.

Samples are incubated with a gelatin or protein substrate under controlled pH and temperature per USP requirements. The extent of protein digestion is determined by titration of released amino groups. Enzyme activity is calculated according to the USP definition of one GDU. Calibration standards and duplicate analyses ensure accuracy and reproducibility.

Testing verifies enzyme potency, supports label claims, and ensures batch-to-batch consistency in bromelain-containing formulations.

This assay quantifies butyric acid (butyrate), a short-chain fatty acid commonly used in gut health and digestive support supplements. Using HPLC, it measures butyrate content in capsules, powders, or coated formulations to verify label claims and support functional product development.

Samples are extracted in aqueous or acidified solvent and analyzed by HPLC with UV or refractive index detection, depending on the matrix. Quantification is performed using certified butyric acid standards, with internal standard correction and duplicate injections to ensure precision.

Results are reported in mg per g or per serving. Values are compared to label claims and formulation targets to confirm proper dosing and detect potential degradation or substitution.

This test confirms the botanical identity of cacao powder (Theobroma cacao) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic methylxanthine and flavanol fingerprint — including theobromine, caffeine, and epicatechin — of the sample is compared against a certified Theobroma cacao reference standard to confirm species authenticity and detect substitution with carob powder, cocoa substitutes, or other botanical fillers. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or ethanol-water and applied alongside a certified cacao reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, examined under UV at 254 nm and 366 nm, and derivatized with an appropriate detection reagent. The resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and fluorescence profile.

Cacao powder is subject to adulteration with carob, roasted grain powders, and other dark-colored botanical materials that visually resemble it. HPTLC identity testing provides a defensible species confirmation based on the unique methylxanthine and flavanol profile of Theobroma cacao, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies caffeine, a widely used stimulant found in energy drinks, pre-workouts, nootropics, and fat burners. Using HPLC, it verifies caffeine content to ensure label accuracy and prevent over- or under-dosing in finished products.

Samples are extracted in aqueous or methanol solution and analyzed by HPLC with UV detection at a caffeine-specific wavelength. Quantification is performed using certified caffeine standards, with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in mg per g or per serving. Values are compared with declared label claims and formulation targets to confirm proper dosing and detect any misformulation.

This assay quantifies calcium beta-hydroxybutyrate (Ca-BHB), a supplemental ketone salt used in ketogenic and performance-enhancing formulations. Using HPLC, it measures BHB content to verify label claims and ensure proper dosing in powders, capsules, and ready-to-mix products.

Samples are extracted in aqueous solution and analyzed by HPLC with UV or refractive index detection, depending on the matrix. Quantification is performed using certified BHB standards, with internal standard correction and duplicate runs to ensure accuracy.

Results are reported in mg per g or per serving. Values are compared to label claims and formulation targets to confirm ketone delivery and detect underdosing or substitution with non-bioavailable salts.

This assay quantifies calcium D-glucarate, a calcium salt of D-glucaric acid used to support liver detox pathways and estrogen metabolism. Using LC-MS/MS, it verifies active content in supplements and functional blends to ensure proper dosing and accurate labeling in detox and hormone-support formulations.

Samples are extracted in aqueous solution and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified calcium D-glucarate standards, with internal standard correction and duplicate runs to ensure accuracy and reproducibility.

Results are reported in mg per g or per serving. Values are compared to label claims and formulation specifications to confirm potency and detect any underdosing or raw material inconsistencies.

This test measures the concentration of Calcium HMB, the calcium salt of β-hydroxy-β-methylbutyrate, to verify the potency and quality of raw materials and finished products such as powders and capsules. The assay uses High-Performance Liquid Chromatography (HPLC) with UV detection to specifically quantify Calcium HMB. Results are reported in mg per gram or per serving to ensure accurate dosage compliance.

Samples are prepared by dissolving a weighed portion in an aqueous solvent, followed by filtration to remove particulates. The solution is injected into an HPLC system equipped with a reversed-phase column and UV detection set at 210 nm, optimal for HMB absorption. Quantification is achieved using a calibration curve generated from certified Calcium HMB reference standards across a defined concentration range. Method accuracy and precision are confirmed through duplicate injections, inclusion of quality control samples, and spike recovery experiments to validate extraction efficiency.

Testing verifies label claims, ensures product consistency, and confirms the standardized potency expected in CaHMB-containing formulations.

This assay measures the concentration of key cannabinoids using High-Performance Liquid Chromatography (HPLC) with UV or diode array detection. The method provides precise separation and quantification of acidic and neutral cannabinoids without heat-induced decarboxylation.

Samples are extracted in organic solvent and filtered prior to HPLC analysis. Cannabinoids are separated under validated chromatographic conditions and quantified against certified reference standards. The method typically covers THC, THCa, CBD, CBDa, CBN, CBG, CBGa, CBDV, THCV, Δ8-THC, and CBC. Calibration curves, retention time confirmation, and replicate injections ensure accuracy and reproducibility.

Results are reported in % w/w (raw material), mg/g, or mg/serving for finished products. Testing confirms compliance with hemp THC limits (e.g., <0.3% Δ9-THC), verifies label claims, and supports potency and quality assurance programs.

This test quantifies caprylic acid (C8:0), a medium-chain saturated fatty acid, in dietary supplements, MCT oil products, and raw materials using High-Performance Liquid Chromatography (HPLC). Caprylic acid is valued for its rapid absorption and conversion to ketones, making it a key active ingredient in MCT-based and ketogenic formulations. Accurate quantification ensures the declared concentration of caprylic acid is present and that the fatty acid profile meets product specifications. Results are reported in mg per serving or as a percentage of total fat content, as applicable.

A representative sample is weighed and subjected to fatty acid extraction using an organic solvent system, followed by derivatization — typically as fatty acid methyl esters (FAMEs) or phenacyl esters — to improve chromatographic retention and UV detectability. The derivatized extract is injected onto a reversed-phase C18 HPLC column, and detection is performed by UV at approximately 210–254 nm depending on the derivatization approach used. Quantification is performed against a multi-point external calibration curve prepared from a certified caprylic acid reference standard, with system suitability and QC samples run concurrently to confirm method performance.

Caprylic acid is a primary label-claimed ingredient in MCT oil and ketogenic supplement products, and accurate potency measurement is essential for label claim substantiation and cGMP compliance. HPLC with derivatization provides the retention and UV sensitivity needed to quantify caprylic acid specifically within a broader fatty acid profile, distinguishing it from capric acid (C10:0) and other co-present medium- and long-chain fatty acids in complex lipid matrices.

This test confirms the botanical identity of cardamom raw materials — including whole or ground fruit, seed, powder, and extract — using High-Performance Thin-Layer Chromatography (HPTLC). True or green cardamom is derived from the dried fruit of Elettaria cardamomum (L.) Maton (Zingiberaceae). It is used in dietary supplements and functional formulations for its aromatic, digestive, and traditional wellness applications. Cardamom contains a characteristic essential-oil profile rich in monoterpenes and oxygenated monoterpenes, including 1,8-cineole and α-terpinyl acetate, together with other volatile and nonvolatile constituents that collectively form its characteristic chemical fingerprint. HPTLC compares the chromatographic profile of the sample with authenticated E. cardamomum reference material and, where suitable, relevant marker standards. The resulting multi-zone fingerprint helps verify the declared botanical source and assess potential substitution, dilution, or admixture with other spice materials, including black cardamom species, ginger-family botanicals, or nonconforming plant material. Identity testing is qualitative and does not determine essential-oil potency, volatile-oil composition, microbial quality, pesticide residues, or heavy-metal content.

A representative sample is accurately weighed and extracted using a validated solvent system selected to recover the characteristic cardamom constituents. A non-polar or moderately polar solvent, such as dichloromethane, ethyl acetate, or an appropriate hydroalcoholic system, may be used depending on whether the method is designed to emphasize volatile oil-associated constituents, nonvolatile constituents, or a combined fingerprint. The clarified extract is applied alongside authenticated Elettaria cardamomum reference material and, when included in the method scope, qualified marker standards such as 1,8-cineole and α-terpinyl acetate, onto a silica gel 60 F₂₅₄ HPTLC plate using an automated sample applicator. The plate is developed in a validated mobile-phase system optimized to resolve the characteristic cardamom fingerprint. After development, the plate is dried, documented under UV light at 254 nm and 366 nm, and visualized after derivatization with an appropriate reagent, such as anisaldehyde-sulfuric acid or vanillin-sulfuric acid, followed by controlled heating. The positions, colors, fluorescence, and relative intensities of characteristic zones are compared with the authenticated reference profile. Identity is confirmed when the sample fingerprint is concordant with the reference material. System-suitability criteria and concurrent quality-control samples are evaluated with each analytical run.

Cardamom is traded in multiple physical forms and can be vulnerable to substitution or dilution, particularly after grinding or extraction, when organoleptic and morphological inspection is no longer sufficient. HPTLC provides a rapid, cost-effective, and information-rich identity method by evaluating the combined chromatographic pattern of characteristic constituents rather than relying on a single marker that may be variable among cultivars, origins, or processed materials. The multi-marker fingerprint strengthens differentiation of authentic E. cardamomum from related spices and alternative cardamom materials with different chemical profiles. This method supports incoming raw-material qualification and identity testing under dietary supplement cGMP requirements in 21 CFR 111.75 and is consistent with the fit-for-purpose HPTLC approach described in USP General Chapter <203>.

This assay quantifies L-carnitine, a conditionally essential nutrient involved in mitochondrial energy transfer and fat metabolism. Using LC-MS/MS, it verifies L-carnitine content in supplements, beverages, and performance products to ensure proper dosing and label accuracy.

Samples are extracted in acidified aqueous solution and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using high-purity L-carnitine standards with internal standard correction and duplicate injections to ensure precision and reproducibility.

Results are reported in mg per g or per serving. Values are compared to declared label claims and formulation targets. This assay helps confirm ingredient integrity and supports functional claims in metabolic health and sports nutrition products.

This test quantifies carnosic acid, the primary bioactive diterpene phenol found in rosemary (Rosmarinus officinalis) and sage extracts, in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Carnosic acid is the principal marker compound used to define the potency and standardization of rosemary extracts, valued for its strong antioxidant activity and studied neuroprotective properties. Results are reported in mg per serving or as a percentage of extract weight to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a methanol or acetonitrile-water solvent system with sonication to ensure complete recovery of carnosic acid from the botanical matrix. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 230 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified carnosic acid reference standard. Sample preparation is conducted under reduced-light conditions where possible to minimize oxidative degradation of this light- and oxygen-sensitive compound, and system suitability and QC samples are run concurrently to confirm method performance.

Carnosic acid is the primary standardization marker for rosemary extract and is highly susceptible to oxidative degradation, meaning its measured concentration is a direct indicator of both extract potency and raw material freshness. Accurate HPLC-UV quantification is essential for verifying that rosemary-based ingredients meet their stated standardization levels and for distinguishing carnosic acid from its oxidation product carnosol and other co-present phenolic compounds in complex botanical matrices.

This test quantifies carvacrol — the primary phenolic monoterpenoid found in oregano (Origanum vulgare) essential oil and a key bioactive compound in thyme and other aromatic herbs — in botanical extracts, essential oils, raw materials, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Carvacrol is the principal marker compound used to standardize oregano oil extracts and is responsible for the ingredient's well-documented antimicrobial, antifungal, and antioxidant activity. Accurate quantification confirms that the extract meets its declared potency and that the correct botanical material is present. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved or diluted in an appropriate organic solvent such as methanol or ethanol. The solution is filtered through a 0.2 µm membrane and analyzed by reversed-phase HPLC on a C18 column, with UV detection at approximately 270–280 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified carvacrol reference standard. Peak identity is confirmed by retention time and UV spectral comparison to the reference standard, and the method resolves carvacrol from its structural isomer thymol, which may co-occur in oregano and thyme-derived materials. System suitability and quality control standards are run concurrently to confirm method accuracy and precision.

Oregano oil extracts are commercially standardized on the basis of carvacrol content, typically to 60–80% or higher, making accurate HPLC quantification essential for verifying that a given extract meets its standardization claim. Carvacrol and thymol are structural isomers with similar UV absorbance profiles, and HPLC provides the chromatographic resolution needed to quantify each compound independently — an important distinction as their relative proportions vary between botanical species and can affect both the biological activity and the authenticity of the extract. This test supports raw material qualification and label claim compliance under 21 CFR 111.

This panel quantifies key catechins commonly found in green tea and botanical extracts: epicatechin, epigallocatechin (EGC), epicatechin gallate (ECG), and epigallocatechin gallate (EGCG). Using HPLC, it verifies the concentration of each catechin to ensure consistency in antioxidant and metabolic health formulations.

Samples are extracted using methanol or aqueous solvents under light-protected conditions. The extract is analyzed by HPLC with UV detection at catechin-specific wavelengths. Quantification is performed using certified standards for each compound, with internal standard correction and duplicate runs to ensure accuracy.

Results are reported in mg per g or per serving for each catechin. Values are compared to formulation targets and label claims to confirm active content and support claims related to antioxidant, cardiovascular, or metabolic benefits.

This test quantifies a panel of individual catechins — including epicatechin (EC), epigallocatechin (EGC), epicatechin gallate (ECG), epigallocatechin gallate (EGCG), catechin (C), and gallocatechin (GC) — in green tea extracts, botanical ingredients, and dietary supplements using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Catechins are the primary bioactive polyphenols in green tea and are associated with antioxidant, anti-inflammatory, and metabolic health benefits, with EGCG recognized as the most potent and extensively studied individual catechin. Individual catechin quantification provides a more complete and specific measure of extract quality and potency than total polyphenol or total catechin assays alone. Results are reported in milligrams per gram or per serving for each individual catechin.

A representative sample is accurately weighed and extracted using aqueous methanol or ethanol, with ascorbic acid added to the extraction solvent to prevent oxidative degradation of the catechins during sample preparation. Isotopically labeled or structurally analogous internal standards are added prior to extraction to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase LC-MS/MS on a C18 column, with detection in negative electrospray ionization (ESI) mode using MRM transitions specific to each individual catechin. Quantification is performed against a multi-point external calibration curve prepared from certified reference standards for each target catechin. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity across the full panel.

Green tea extracts are commercially standardized on the basis of total catechin or EGCG content, and individual catechin profiling by LC-MS/MS provides the most specific and comprehensive measure of extract quality, authenticity, and potency. Total polyphenol or UV-based catechin assays do not distinguish between individual catechins and can be confounded by non-catechin polyphenols, whereas LC-MS/MS with MRM detection provides compound-specific quantification of each catechin at the sensitivity levels required for complex botanical matrices. This level of analytical detail is essential for premium green tea ingredient qualification, EGCG label claim substantiation, and detection of adulteration or dilution with low-catechin materials under 21 CFR 111.

This test identifies and characterizes Ceylon cinnamon (Cinnamomum verum, syn. C. zeylanicum) in raw materials, powders, extracts, and dietary supplements using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Ceylon cinnamon — often referred to as "true cinnamon" — is botanically and chemically distinct from the more common cassia varieties (Cinnamomum cassia, C. aromaticum, C. loureiroi), which contain significantly higher levels of coumarin, a naturally occurring compound associated with hepatotoxicity at elevated intakes. LC-MS/MS enables the simultaneous profiling and quantification of key marker compounds — including cinnamaldehyde, cinnamic acid, eugenol, and coumarin — that together constitute a discriminating chemical signature for C. verum authentication and differentiation from cassia adulterants. Results are reported as concentrations of individual marker compounds in milligrams per gram or as a qualitative identity confirmation.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., aqueous methanol or ethanol) to capture the characteristic volatile and non-volatile phytochemical profile of C. verum. An isotopically labeled internal standard is added prior to extraction to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase LC-MS/MS using electrospray ionization (ESI) in positive and/or negative ion mode, with multiple reaction monitoring (MRM) transitions selected for key marker compounds including cinnamaldehyde, cinnamic acid, eugenol, and coumarin. Quantification is performed against multi-point external calibration curves prepared from certified reference standards for each target compound. The resulting marker compound profile is compared to authenticated C. verum reference data to confirm species identity and assess the presence of cassia adulteration. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity.

The primary driver for LC-MS/MS-based authentication of Ceylon cinnamon is the critical distinction between C. verum and cassia varieties based on coumarin content. Ceylon cinnamon contains very low levels of coumarin (typically < 0.01% w/w), whereas cassia varieties contain substantially higher levels (0.3–1.0% w/w or greater), which is of regulatory significance in markets such as the EU where coumarin in food supplements is subject to maximum level restrictions. LC-MS/MS provides the sensitivity and specificity to simultaneously quantify coumarin at trace levels alongside other marker compounds, enabling definitive species authentication and adulteration detection that cannot be achieved by HPTLC or single-marker HPLC methods alone. This supports label claim accuracy, raw material qualification, consumer safety, and cGMP compliance under 21 CFR 111.

This test confirms the identity of chaga mushroom (Inonotus obliquus) in raw materials, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Chaga is a parasitic fungus found predominantly on birch trees in northern climates and is widely used in dietary supplements for its antioxidant, immune-modulating, and adaptogenic properties. Its characteristic phytochemical profile includes betulinic acid, inotodiol, ergosterol, and melanin-like pigments derived from the birch host. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated reference standard to confirm species identity and detect potential adulteration or substitution with inferior or unrelated fungal materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or ethanol) to capture the characteristic secondary metabolite profile of I. obliquus. The extract is applied alongside a certified chaga reference standard and, where applicable, potential adulterant extracts, onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve chaga's characteristic marker compounds, including betulinic acid and inotodiol. After development, the plate is derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid or vanillin-sulfuric acid) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with HPTLC identity testing guidelines.

Chaga mushroom is subject to adulteration and misidentification, particularly given the premium pricing of authenticated Inonotus obliquus material and the potential for substitution with other conk fungi or non-chaga birch-associated species. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that is more discriminating than single-marker assays, enabling detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test confirms the identity of chanca piedra (Phyllanthus niruri L., and related pharmacopeial species including P. amarus and P. urinaria) in raw materials, whole plant powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Phyllanthus niruri, commonly known as chanca piedra ("stone breaker"), gale of the wind, or seed-under-leaf, is a tropical herb with a long history of use in Ayurvedic, Amazonian, and traditional Chinese medicine for supporting kidney stone management, urinary tract health, and liver function. Its characteristic phytochemical profile includes lignans (phyllanthin, hypophyllanthin, niranthin), ellagitannins (geraniin, corilagin), flavonoids (quercetin, rutin, astragalin), and alkaloids (phyllanthine, securinine). HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated P. niruri reference standard to confirm species identity and detect potential adulteration, substitution with other Phyllanthus species, or blending with unrelated plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic secondary metabolite profile of P. niruri, including lignans, tannins, and flavonoids. The extract is applied alongside a certified P. niruri reference standard and, where applicable, potential adulterant or related species extracts (e.g., P. amarus, P. urinaria), onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve the characteristic marker compounds of P. niruri. After development, the plate is derivatized with an appropriate reagent (e.g., Natural Products Reagent A / NP/PEG for flavonoid and lignan visualization, or anisaldehyde-sulfuric acid) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Phyllanthus niruri is subject to adulteration and species substitution, as several closely related Phyllanthus species share morphological similarities and overlapping geographic distribution, making visual identification of dried and powdered materials unreliable. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation — anchored by the characteristic lignan and tannin profile — that is more discriminating than single-marker assays, enabling detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test confirms the identity of chasteberry (Vitex agnus-castus L.) in raw materials, fruit powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Vitex agnus-castus, commonly known as chasteberry or chaste tree, is a well-established botanical used in women's health formulations for its traditionally recognized role in supporting hormonal balance, menstrual regularity, and premenstrual syndrome (PMS) symptom relief. Its characteristic phytochemical profile includes iridoid glycosides (notably agnuside and aucubin), flavonoids (casticin, luteolin, apigenin), and diterpenes. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated V. agnus-castus reference standard to confirm species identity and detect potential adulteration, substitution, or misidentification with other Vitex species or unrelated plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic secondary metabolite profile of V. agnus-castus, including iridoid glycosides and flavonoids. The extract is applied alongside a certified chasteberry reference standard and, where applicable, potential adulterant extracts, onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated solvent system optimized to resolve the characteristic marker compounds of V. agnus-castus, including agnuside and casticin. After development, the plate is derivatized with an appropriate reagent (e.g., Natural Products Reagent A / NP/PEG for flavonoid visualization, or anisaldehyde-sulfuric acid) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting fingerprint pattern is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Chasteberry is subject to adulteration and species substitution, including replacement with other Vitex species that may share morphological similarities but differ significantly in phytochemical composition and biological activity. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that is more discriminating than single-marker assays, enabling detection of substitution or adulteration that would not be apparent from potency testing of individual marker compounds alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test confirms the identity of chlorella — primarily Chlorella vulgaris Beyerinck and Chlorella pyrenoidosa H.Chick, unicellular green microalgae (Chlorophyta) widely used in dietary supplements — in raw materials, dried powders, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Chlorella is one of the most commercially significant microalgae-based dietary supplements globally, valued for its high chlorophyll content, complete protein profile, and Chlorella Growth Factor (CGF). Its characteristic phytochemical profile includes chlorophylls a and b, carotenoids (lutein, β-carotene, zeaxanthin), and tocopherols. HPTLC identity testing generates a characteristic pigment-based chromatographic fingerprint that is compared against an authenticated chlorella reference standard to confirm species identity and detect potential adulteration, substitution with other microalgae (e.g., spirulina/Arthrospira spp.), or blending with non-algal plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., acetone or methanol) to capture the characteristic pigment profile of chlorella, including chlorophylls a and b and carotenoids. The extract is applied alongside a certified chlorella reference standard and, where applicable, potential adulterant extracts (e.g., spirulina powder, other green microalgae), onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated non-polar solvent system optimized to resolve the characteristic pigment bands of chlorella, including chlorophyll a, chlorophyll b, lutein, and β-carotene. After development, the plate is evaluated under white light and UV light at 254 nm and 366 nm, where the characteristic green and yellow-orange pigment bands of chlorella are visualized without derivatization. The resulting fingerprint pattern — notably the presence of both chlorophyll a and chlorophyll b, and the characteristic carotenoid profile including lutein — is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

Chlorella is subject to adulteration and species substitution, including blending with spirulina, other microalgae, or non-algal plant powders. A key distinguishing feature of chlorella relative to spirulina is the presence of both chlorophyll a and chlorophyll b (spirulina contains chlorophyll a but not chlorophyll b) and a distinct carotenoid profile enriched in lutein rather than zeaxanthin. HPTLC pigment fingerprinting exploits these characteristic compositional differences to provide a discriminating identity confirmation that distinguishes authentic chlorella from spirulina and other potential substitutes. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test measures the total chlorogenic acids, including key caffeoylquinic acid isomers (3-CQA, 4-CQA, and 5-CQA), which are important antioxidants in plant-based raw materials and finished products. Using High-Performance Liquid Chromatography (HPLC) with UV detection, the assay quantifies these compounds to ensure ingredient potency and consistency. Results are reported in milligrams per gram (mg/g) or as a percentage of the sample weight, supporting quality control and standardization efforts.

Samples are prepared by extracting 0.5 grams of powdered material with 10 mL of 70% methanol using sonication for 30 minutes, followed by filtration. The extract is injected into an HPLC system equipped with a C18 reversed-phase column and UV detection at 325 nm. Separation of 3-CQA, 4-CQA, and 5-CQA is achieved using a gradient elution of water and acetonitrile with 0.1% formic acid over 30 minutes. Quantification is performed by comparing peak areas to a calibration curve constructed from certified chlorogenic acid standards. Method accuracy is verified through duplicate injections, spiked recovery tests, and analysis of quality control samples.

Results are reported in mg/g (raw materials) or mg/serving (finished products). Testing verifies standardized potency, ensures product consistency, and supports antioxidant and functional food claims.

This assay measures chlorophyll and related chlorophyll derivatives using High-Performance Liquid Chromatography (HPLC). The method provides accurate quantification of chlorophyll pigments, supporting quality control for plant-based and antioxidant formulations.

Samples are extracted under light-protected conditions and analyzed using validated HPLC chromatographic methods. Chlorophyll a, chlorophyll b, and potential derivatives (e.g., pheophytins) are separated and detected by UV or diode-array detection. Quantitation is performed using certified reference standards with calibration and replicate runs ensuring accuracy and reproducibility.

Testing confirms standardized potency, verifies label claims, and supports consistency in green superfood and botanical products.

This assay quantifies cholesterol content in food and supplement samples using Gas Chromatography (GC) following AOAC Official Method 976.26. It ensures compliance with nutritional labeling requirements and verifies formulation accuracy in animal-based products and fortified foods.

Samples are saponified to release cholesterol from esters, then extracted with organic solvents. The extract is analyzed by Gas Chromatography with Flame Ionization Detection (GC-FID). Quantification is achieved using calibration curves from high-purity cholesterol standards, with duplicate runs and internal reference correction to ensure precision.

Results are reported in mg per 100 g or per serving. The values are compared with regulatory thresholds and label claims. Accurate cholesterol quantification supports transparency in nutrition facts panels and helps monitor lipid levels in dietary and functional formulations.

This assay quantifies choline—an essential nutrient involved in liver function, brain development, and cell membrane structure—in food and supplement matrices. It measures both free choline and that released from bound forms after conversion. The method employs LC‑MS/MS for high sensitivity and accuracy, supporting nutritional labeling and product quality assessment.

The sample is subjected to an extraction process (often including acid digestion) to liberate choline from bound compounds. An internal standard is added to correct for any losses during processing. The extract is analyzed by LC‑MS/MS, with calibration performed using choline standards and quality control measures ensuring data integrity.

Results are provided in mg per 100 g or per serving. The reported levels allow comparison with nutritional targets and formulation claims. Consistency across batches confirms reliable manufacturing, while deviations may prompt adjustments in fortification levels.

This assay measures choline derived from choline bitartrate using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high sensitivity and selectivity for accurate quantification in raw materials and finished products.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Choline is detected using multiple reaction monitoring (MRM) and quantified against certified reference standards. Internal calibration and quality control checks ensure accuracy and reproducibility across matrices.

Testing verifies label claims, confirms ingredient purity, and ensures batch-to-batch consistency in choline-containing formulations.

This assay measures chondroitin sulfate, a sulfated glycosaminoglycan commonly sourced from bovine, porcine, or marine cartilage. HPLC analysis provides accurate quantification of total chondroitin content to ensure standardized potency in raw materials and finished formulations.

Samples are enzymatically or chemically prepared to release chondroitin sulfate into measurable fragments, then analyzed under validated HPLC chromatographic conditions. Chondroitin sulfate is separated from other glycosaminoglycans and detected via UV or refractive index detection. Quantitation is performed using certified reference standards, with calibration curves and duplicate injections ensuring accuracy and reproducibility.

Testing verifies label claims, confirms raw material authenticity, detects adulteration, and ensures consistency in joint-support formulations.

This test confirms the botanical identity of cinnamon (Cinnamomum verum or C. cassia) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). HPTLC is the industry-standard method for botanical identity verification, producing a characteristic fingerprint chromatogram that is compared against a certified reference standard to confirm species authenticity and rule out substitution or adulteration with inferior cinnamon species or unrelated botanicals. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using an appropriate organic solvent and applied alongside a certified cinnamon reference standard onto an HPTLC silica gel plate using an automated applicator. The plate is developed in a validated solvent system, dried, and derivatized with a detection reagent such as anisaldehyde-sulfuric acid to visualize the characteristic marker compounds. The resulting chromatographic fingerprint of the sample is compared visually and by densitometric scanning to the reference standard pattern. Identity is confirmed when the sample fingerprint matches the reference in terms of Rf values, band positions, and color profile, per established HPTLC identity methods (e.g., AHPA or Ph. Eur. botanical monographs).

Cinnamon is one of the most commonly adulterated botanical ingredients, with true Ceylon cinnamon (C. verum) frequently substituted with the less expensive Cassia cinnamon (C. cassia) or other Cinnamomum species. HPTLC identity testing provides a rapid, visual, and scientifically defensible confirmation of botanical species, supporting supplier qualification, label accuracy, and compliance with cGMP identity testing requirements under 21 CFR 111.

This assay quantifies key bioactive compounds in Cistanche species—primarily echinacoside and acteoside—which are associated with energy, libido, and neuroprotective effects. Using LC-MS/MS, it verifies the potency and standardization of Cistanche extracts in vitality, cognitive, and anti-aging supplements.

Samples are extracted using alcohol-based solvents and analyzed by LC-MS/MS with compound-specific mass transitions. Quantification is performed using certified standards with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm consistent bioactive delivery and detect raw material variability or degradation.

This test confirms the botanical identity of Cistanche tubulosa in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic phenylethanoid glycoside fingerprint — including echinacoside and acteoside — of the sample is compared against a certified Cistanche tubulosa reference standard to confirm species authenticity and detect substitution with related Cistanche species or other botanical materials. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or ethanol-water and applied alongside a certified Cistanche tubulosa reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, derivatized with anisaldehyde-sulfuric acid or natural products reagent, and the fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Cistanche tubulosa is a premium botanical with a risk of substitution with the less expensive Cistanche deserticola or other Cistanche species that differ in their phenylethanoid glycoside profile. HPTLC identity testing provides a rapid and defensible species confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies citicoline (CDP-choline), a bioavailable choline donor used in nootropic and neurological support supplements. Using HPLC, it measures citicoline content in raw materials and finished formulations to verify potency, ensure label accuracy, and detect degradation or adulteration.

Samples are extracted in aqueous or buffered solvent and analyzed by HPLC with UV detection at a compound-specific wavelength (typically ~270 nm). Quantification is performed using certified citicoline standards, with internal standard correction and duplicate injections to ensure reproducibility and accuracy.

Results are reported in mg per g or per serving. Values are compared against formulation targets and declared label claims to confirm dosing accuracy, validate standardized potency, and ensure consistency across production batches.

This test quantifies citric acid content in raw materials, powders, and finished products using High-Performance Liquid Chromatography (HPLC). Citric acid is important for flavor enhancement, preservation, and pH regulation in formulations. The method provides precise measurement down to 0.01% concentration, supporting quality control and regulatory compliance.

Samples are prepared by dissolving a representative portion in deionized water followed by filtration to remove particulates. The extract is injected into an HPLC system equipped with a C18 reversed-phase column and detected using UV absorbance at 210 nm. Quantification is achieved by comparing peak areas to a calibration curve generated from certified citric acid reference standards. Method accuracy is verified through duplicate injections and spike recovery tests to ensure reproducibility and reliability.

Testing verifies correct formulation levels, supports quality assurance, and ensures consistent sensory and functional performance.

This assay quantifies cocoa flavonols using High-Performance Liquid Chromatography (HPLC). The method provides accurate profiling of cocoa polyphenols to assess purity and active content.

Samples are extracted and analyzed by HPLC under validated chromatographic conditions. Major flavonol compounds—such as catechin, epicatechin, and procyanidins—are separated and detected via UV or fluorescence detection. Quantitation is performed against certified standards, with duplicate runs ensuring precision.

Results are reported as mg/g (raw material) or mg/serving (finished products). Testing verifies standardized potency in cocoa extracts, supports antioxidant and cardiovascular health claims, and ensures batch-to-batch consistency.

This test confirms the botanical identity of coconut (Cocos nucifera) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic lipid and phenolic fingerprint of the sample is compared against a certified Cocos nucifera reference standard to confirm botanical authenticity and detect substitution with other palm-derived or vegetable oil materials. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using an appropriate organic solvent and applied alongside a certified Cocos nucifera reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, derivatized with an appropriate detection reagent, and the fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Coconut-derived ingredients — including coconut powder, coconut oil, and MCT fractions — are subject to substitution with other palm-derived oils and materials. HPTLC identity testing provides a botanical-level confirmation of species authenticity, supporting supplier qualification, allergen declaration accuracy, and cGMP compliance under 21 CFR 111.

This test quantifies total collagen content in collagen raw materials, hydrolysates, and dietary supplements by measuring hydroxyproline — a non-standard amino acid formed by post-translational hydroxylation of proline and found almost exclusively in collagen and elastin — using High-Performance Liquid Chromatography (HPLC). Hydroxyproline constitutes approximately 13–14% of the total amino acid composition of collagen by mass, making it the most specific and reliable chemical marker for collagen quantification. Total collagen content is calculated from the measured hydroxyproline concentration using the established conversion factor (collagen content = hydroxyproline content × 7.14, based on the mean hydroxyproline content of collagen). This approach is widely used for quality control of collagen peptide, gelatin, and collagen hydrolysate ingredients across the food, pharmaceutical, and dietary supplement industries. Results are reported as a percentage or in milligrams of collagen per gram or per serving.

A representative sample is accurately weighed and subjected to complete acid hydrolysis using 6N hydrochloric acid at elevated temperature (typically 110°C for 22–24 hours under reflux or sealed-vessel conditions) to fully hydrolyze the collagen protein into its constituent free amino acids, including hydroxyproline. The hydrolysate is neutralized, filtered, and subjected to pre-column derivatization — commonly using o-phthalaldehyde (OPA) or 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) — to generate UV- or fluorescence-detectable hydroxyproline derivatives. The derivatized hydrolysate is analyzed by reversed-phase HPLC on a C18 column with UV or fluorescence detection, providing chromatographic resolution of hydroxyproline from other amino acids present in the hydrolysate. Quantification is performed against a multi-point external calibration curve prepared from a certified trans-4-hydroxy-L-proline reference standard. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Hydroxyproline is the most specific and widely accepted chemical marker for collagen quantification because it is found at high and consistent concentrations in collagen and is present at negligible levels in virtually all other proteins. This specificity makes hydroxyproline-based HPLC quantification a more accurate and collagen-specific measure of collagen content than total protein assays (e.g., Kjeldahl or Bradford), which cannot distinguish collagen from other co-occurring proteins. HPLC with derivatization provides the sensitivity and chromatographic resolution required to accurately quantify hydroxyproline in complex collagen hydrolysate and finished supplement matrices. This method supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This test identifies and quantifies collagen Types I, II, and III — the three most prevalent collagens in mammals, each with distinct tissue distributions and functional roles — in collagen raw materials, hydrolysates, and dietary supplements using enzymatic hydrolysis followed by Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Type I collagen is the most abundant structural protein in bone, skin, and tendons; Type II is the primary collagen of articular cartilage and is used in joint health formulations; and Type III is found alongside Type I in skin, blood vessels, and connective tissue. LC-MS/MS-based collagen typing via signature peptide analysis provides definitive, species-independent identification and quantification of each collagen type, supporting label claim verification, raw material authentication, and detection of collagen source adulteration. Results are reported as a percentage or in milligrams per gram or per serving for each collagen type.

A representative sample is accurately weighed and subjected to controlled enzymatic hydrolysis using a combination of sequence-specific proteases (e.g., trypsin, chymotrypsin, or collagenase) under optimized conditions to generate reproducible peptide fragments characteristic of each collagen type. The resulting peptide digest is desalted, filtered, and analyzed by reversed-phase LC-MS/MS using electrospray ionization (ESI) in positive ion mode. Collagen types are identified and quantified using multiple reaction monitoring (MRM) of type-specific signature peptides — including hydroxyproline-containing peptides unique to collagen — derived from the characteristic triple-helical domain sequences of Types I, II, and III collagen. Quantification is performed against isotopically labeled internal standards or certified peptide reference standards for each collagen type. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity.

Collagen types cannot be distinguished by total protein assays, amino acid analysis, or standard HPLC methods alone, as they share a common amino acid composition dominated by glycine, proline, and hydroxyproline. LC-MS/MS with signature peptide analysis provides the only method capable of simultaneously identifying and quantifying individual collagen types with the specificity required to confirm source authenticity (e.g., bovine, porcine, marine, or chicken), detect adulteration or substitution, and verify that the declared collagen type is present at the stated level. This approach is aligned with established proteomics-based methods for collagen characterization and supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies conjugated linoleic acid (CLA), a group of omega-6 fatty acid isomers commonly found in weight management and sports nutrition supplements. Using LC-MS/MS, it verifies CLA content in oils, softgels, and powders to confirm label claims and ensure proper dosing in functional products.

Samples are extracted in organic solvent and analyzed by LC-MS/MS using isomer-specific mass transitions. Quantification is performed with certified CLA standards, internal standard correction, and duplicate injections to ensure accurate and reproducible results.

Results are reported in mg per g or per serving. Values are compared to label claims and formulation targets to confirm consistency, detect degradation, or identify formulation issues across batches.

This test quantifies Coenzyme Q10 (ubiquinone) — a fat-soluble, endogenously produced compound essential for mitochondrial ATP synthesis and cellular antioxidant protection — in dietary supplements, raw materials, and finished products using High-Performance Liquid Chromatography (HPLC). CoQ10 is one of the most widely used ingredients in cardiovascular and energy support supplements, and accurate potency verification is critical for label claim substantiation given the significant cost of pharmaceutical-grade CoQ10 and its susceptibility to underdosing. This method is applicable to both the oxidized form (ubiquinone) and, with appropriate method adaptation, the reduced form (ubiquinol). Results are reported as a percentage or in milligrams per serving.

A representative sample is accurately weighed and extracted using an organic solvent such as ethanol, hexane, or a methanol/isopropanol mixture, with sonication to ensure complete dissolution of the lipophilic CoQ10. The extract is filtered through a 0.2 µm membrane and analyzed by reversed-phase HPLC on a C18 column, with UV or Electrochemical (ECD) detection at approximately 275 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified CoQ10 (ubiquinone) reference standard. System suitability and quality control standards are run concurrently to confirm method accuracy and precision, and all sample preparation steps are performed with minimal light exposure to prevent photodegradation.

CoQ10 is a high-value ingredient that commands a significant price premium, making it one of the more commonly underdosed actives in the supplement industry. HPLC with UV or electrochemical detection provides the sensitivity and specificity needed to accurately quantify CoQ10 in complex lipid-rich matrices, distinguishing it from structurally related ubiquinol and other fat-soluble co-extractives. Rigorous potency testing is essential for label claim compliance, raw material qualification, and consumer trust in cardiovascular and energy support formulations under 21 CFR 111.

This test quantifies cordycepic acid (D-mannitol) — a primary marker compound found in Cordyceps species, including Cordyceps sinensis and Cordyceps militaris — in mushroom extracts, raw materials, and dietary supplements using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Cordycepic acid is used alongside other markers such as cordycepin and adenosine to authenticate Cordyceps identity and verify extract potency. Due to the widespread use of mycelium-on-grain products and synthetic substitutes in the market, accurate quantification of genuine marker compounds is critical for quality assurance and label claim substantiation. Results are reported as a percentage or in milligrams per gram.

A representative sample is accurately weighed and extracted using an aqueous solvent or dilute methanol, with sonication or agitation to ensure complete dissolution of the polar marker compounds. The extract is filtered through a 0.2 µm membrane and analyzed by LC-MS/MS using a hydrophilic interaction liquid chromatography (HILIC) or reversed-phase C18 column. Detection is performed in positive or negative electrospray ionization (ESI) mode, with multiple reaction monitoring (MRM) transitions selected for cordycepic acid to provide high specificity and sensitivity. Quantification is performed against a multi-point external calibration curve prepared from a certified D-mannitol or cordycepic acid reference standard. Internal standards and quality control samples are run concurrently to confirm method accuracy and precision.

The Cordyceps supplement market is heavily affected by adulteration, with many products containing mycelium grown on grain substrates that contribute negligible levels of the bioactive compounds present in authentic fruiting body extracts. LC-MS/MS provides the sensitivity and specificity needed to accurately quantify cordycepic acid at low concentrations in complex mushroom matrices, distinguishing genuine Cordyceps material from grain-based fillers and synthetic substitutes. This test is a critical component of a comprehensive Cordyceps quality panel alongside cordycepin and adenosine quantification, supporting both supplier qualification and finished product release under 21 CFR 111.

This test quantifies cordycepin (3'-deoxyadenosine) — a naturally occurring adenosine analog and the principal bioactive nucleoside constituent of Cordyceps militaris — in raw materials, fungal extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Cordycepin is one of the most pharmacologically significant compounds in the Cordyceps genus, with a growing body of research supporting its roles in energy metabolism, immune modulation, anti-inflammatory activity, and cellular health. It is used as a key potency marker to differentiate high-quality C. militaris extracts from Cordyceps sinensis and mycelium-based products, which typically contain little to no cordycepin. Accurate quantification is essential for label claim substantiation and for confirming the quality and authenticity of Cordyceps raw materials. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and extracted using an appropriate aqueous solvent system (e.g., water or aqueous methanol) to ensure complete dissolution of cordycepin and related nucleosides. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 260 nm, the characteristic absorption maximum of the adenine nucleobase chromophore shared by cordycepin and adenosine. Quantification is performed against a multi-point external calibration curve prepared from a certified cordycepin reference standard. Where applicable, related nucleosides — including adenosine and cordycepic acid (D-mannitol) — may be monitored simultaneously to provide a broader nucleoside profile and confirm the characteristic Cordyceps composition. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Cordycepin's adenine nucleobase chromophore provides strong UV absorption at 260 nm, making HPLC with UV detection a sensitive and specific method for its quantification without the need for derivatization. Chromatographic separation on a C18 column resolves cordycepin from adenosine and other co-occurring nucleosides based on differences in hydrophobicity, ensuring that potency results reflect cordycepin content specifically. This distinction is analytically critical, as cordycepin and adenosine differ by only a single hydroxyl group at the 3' position of the ribose sugar and cannot be distinguished by UV spectrophotometry alone. The method supports raw material qualification, label claim substantiation, and cGMP compliance under 21 CFR 111.

This test confirms the botanical identity of Cordyceps sinensis mushroom in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic nucleoside fingerprint — including adenosine and cordycepin — of the sample is compared against a certified Cordyceps sinensis reference standard to confirm species authenticity and detect substitution with Cordyceps militaris, mycelium-on-grain products, or unrelated fungal materials. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol-water and applied alongside a certified Cordyceps sinensis reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, examined under UV at 254 nm and 366 nm, and the fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and fluorescence profile.

Cordyceps is one of the most adulterated mushroom ingredients in the supplement market, with mycelium-on-grain products and Cordyceps militaris frequently substituted for true Cordyceps sinensis fruiting body. HPTLC identity testing provides a practical and defensible method for confirming species authenticity, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This test quantifies corosolic acid, the primary bioactive triterpenoid found in banaba leaf (Lagerstroemia speciosa) extract, in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Corosolic acid is the key standardization marker for banaba leaf extract and has been studied for its role in glucose transport and blood sugar regulation. Results are reported in mg per serving or as a percentage of extract weight to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a methanol or ethanol-water solvent system with sonication to ensure complete recovery of corosolic acid from the botanical matrix. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 210 nm, reflecting the limited UV chromophore of this triterpenoid compound, and quantification is performed against a multi-point external calibration curve prepared from a certified corosolic acid reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Corosolic acid is the defining potency marker for banaba leaf extract, and its accurate quantification is essential for verifying that raw materials and finished products meet their declared standardization levels. Detection at 210 nm requires careful mobile phase selection and baseline management to achieve adequate sensitivity and specificity for this low-UV-absorbing triterpenoid in complex botanical matrices, making method validation and QC oversight particularly important for reliable results.

This assay measures cranberry-specific marker compounds using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). Analysis typically targets characteristic cranberry polyphenols, including proanthocyanidins (PACs) and related metabolites, to confirm ingredient identity and standardized potency.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Target cranberry markers are detected using multiple reaction monitoring (MRM) and quantified against certified reference standards. Internal calibration and quality controls ensure accuracy and reproducibility across botanical matrices.

Testing verifies cranberry authenticity, supports label claims (e.g., PAC content), and ensures consistency across batches for urinary health formulations.

This test confirms the botanical identity of cranberry (Vaccinium macrocarpon Aiton) in raw materials, extracts, and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic polyphenolic fingerprint of the sample — including anthocyanins, proanthocyanidins, and flavonols specific to Vaccinium macrocarpon — is compared against a certified cranberry reference standard to confirm species authenticity and detect substitution with other Vaccinium species, grape skin extracts, or synthetic colorants used to mimic cranberry's characteristic profile. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using aqueous methanol or acidified ethanol and applied alongside a certified Vaccinium macrocarpon reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system and examined under UV at 254 nm and 366 nm, and optionally derivatized with a suitable spray reagent such as Natural Products Reagent A (NP/PEG) to enhance visualization of the flavonoid and anthocyanin bands. The resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and fluorescence profile.

Cranberry is a high-demand botanical ingredient that is frequently adulterated with lower-cost fruit extracts — including grape, elderberry, and bilberry — or diluted with non-specific polyphenol sources that lack the A-type proanthocyanidins characteristic of genuine Vaccinium macrocarpon. HPTLC identity testing based on the cranberry polyphenolic fingerprint provides a rapid and defensible species confirmation, supporting supplier qualification, consumer safety, and cGMP compliance under 21 CFR 111.