Phytochemical, Vitamin, and Actives testing

This assay quantifies Rebaudioside A, the main high-intensity sweet glycoside found in stevia (Stevia rebaudiana). Using LC-MS/MS, it verifies the concentration of Rebaudioside A in natural sweetener blends, beverages, and flavored supplements—ensuring product consistency, sweetening power, and protection against sugar adulteration.

Samples are extracted in water or ethanol depending on matrix type, then filtered and injected into an LC-MS/MS system. The method uses compound-specific mass transitions for Rebaudioside A and quantifies against high-purity reference standards. Internal standards and duplicate runs are used for precision and accuracy.

Results are reported in mg/g or mg/serving. Values are assessed against formulation targets, label claims, and stevia standardization guidelines. This test confirms proper sweetener dosing and helps verify that “naturally sweetened” products contain authentic stevia—not hidden added sugars or synthetic sweeteners.

This test quantifies recombinant human lactoferrin (rhLF) — a bioengineered form of human lactoferrin produced through recombinant DNA technology in expression systems such as rice (Oryza sativa), Aspergillus niger, or other approved host organisms — in raw materials and dietary supplements using High-Performance Liquid Chromatography (HPLC). Lactoferrin is an 80 kDa iron-binding glycoprotein naturally present in human milk, saliva, tears, and mucosal secretions, recognized for its multifunctional roles in iron absorption, antimicrobial defense, immune modulation, and gut health. Recombinant human lactoferrin is structurally identical to native human lactoferrin and is used in dietary supplements as a functionally equivalent alternative to bovine lactoferrin, with particular relevance for infant nutrition and sensitive populations. Accurate potency quantification is essential for label claim substantiation and for confirming the concentration of the recombinant protein in the finished product. Results are reported in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate aqueous buffer (e.g., phosphate-buffered saline or sodium acetate buffer at physiological pH) to ensure complete dissolution of the lactoferrin protein. The dissolved sample is filtered and analyzed by reversed-phase HPLC or size-exclusion HPLC (SE-HPLC) on an appropriate column — a C4 or C8 reversed-phase column for protein quantification, or a calibrated size-exclusion column for molecular weight confirmation and purity assessment — with UV detection at 280 nm, the characteristic absorption wavelength of aromatic amino acid residues (tryptophan and tyrosine) in proteins. Quantification is performed against a multi-point external calibration curve prepared from a certified recombinant human lactoferrin reference standard of known protein concentration. Purity assessment by SE-HPLC provides additional information on the relative proportion of intact lactoferrin monomer versus aggregates or degradation products. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

HPLC with UV detection at 280 nm provides a sensitive and specific method for lactoferrin protein quantification based on the characteristic UV absorbance of its aromatic amino acid content, offering greater specificity than total protein assays (e.g., Bradford or BCA) that cannot distinguish lactoferrin from co-formulated proteins or excipients. Reversed-phase HPLC enables quantification of the intact lactoferrin protein, while SE-HPLC provides complementary information on protein integrity, aggregation state, and purity — all critical quality attributes for a recombinant protein ingredient. For recombinant human lactoferrin specifically, HPLC-based quantification and purity assessment supports confirmation that the recombinant protein is present at the declared concentration and in the expected intact form, supporting label claim substantiation and cGMP compliance under 21 CFR 111.

This test quantifies recombinant human lactoferrin (rhLF) — a bioidentical form of the naturally occurring iron-binding glycoprotein produced via precision fermentation — in dietary supplements and raw materials using High-Performance Liquid Chromatography (HPLC). Lactoferrin plays a key role in innate immune defense, antimicrobial activity, iron metabolism, and gut health. Accurate quantification of rhLF is essential for label claim verification and for confirming that the declared potency of this high-value ingredient is present in the finished product. Results are reported in mg per serving or mg per gram to support cGMP compliance.

A representative sample is dissolved or diluted in an appropriate aqueous buffer and filtered prior to injection. The sample is analyzed by size-exclusion HPLC (SE-HPLC) or reversed-phase HPLC (RP-HPLC) depending on the validated method, with UV detection at approximately 280 nm to monitor protein absorbance. Quantification is performed against a multi-point external calibration curve prepared from a certified rhLF reference standard. Peak identity is confirmed by retention time comparison to the reference standard, and system suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Recombinant human lactoferrin is a premium, high-cost ingredient produced through precision fermentation, making accurate potency verification critical for both label claim substantiation and raw material qualification. HPLC provides the resolution needed to quantify rhLF as a discrete protein peak, distinguishing it from other proteins, excipients, and fermentation-derived co-products that may be present in the matrix, ensuring that the declared concentration of this bioactive ingredient is confirmed in every batch.

This test confirms the botanical identity of Reishi mushroom (Ganoderma lucidum) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic triterpenoid and polysaccharide fingerprint of the sample is compared against a certified Ganoderma lucidum reference standard to confirm species authenticity and detect substitution with other Ganoderma species or unrelated fungal 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 Ganoderma lucidum reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, derivatized with anisaldehyde-sulfuric acid 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.

Reishi is among the most adulterated mushroom ingredients in the supplement market, with other Ganoderma species and mycelium-on-grain preparations frequently substituted for authentic fruiting body material. HPTLC identity testing provides a rapid and defensible species confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies resveratrol, a polyphenolic compound primarily found in grapes and Japanese knotweed. Using HPLC, it verifies resveratrol content in supplements, functional foods, and botanical extracts to confirm label claims and ensure consistent dosing in longevity and heart health formulations.

Samples are extracted using alcohol-based solvents under light-protected conditions and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified resveratrol standards, with internal standard correction and duplicate injections to ensure accuracy.

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

This test confirms the botanical identity of Rhodiola rosea in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic rosavins and salidroside fingerprint of the sample is compared against a certified Rhodiola rosea reference standard to confirm species authenticity and detect substitution with other Rhodiola species — such as R. crenulata — that lack the rosavin compounds unique to R. rosea. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or ethanol-water and applied alongside a certified Rhodiola rosea 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.

Rhodiola rosea is one of the most commonly adulterated adaptogens, with R. crenulata — which contains salidroside but not rosavins — frequently substituted due to its lower cost. HPTLC identity testing is the most practical method for distinguishing R. rosea from other Rhodiola species based on the presence of species-specific rosavin compounds, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This test confirms the identity of Chinese rhubarb (Rheum palmatum L., and related pharmacopeial species including R. officinale and R. tanguticum) in raw materials, root powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Rheum palmatum, commonly known as Chinese or medicinal rhubarb, is a well-established botanical used in traditional Chinese medicine and Western herbal practice for its role in supporting digestive function, bowel regularity, and gastrointestinal health. Its characteristic phytochemical profile includes anthraquinone glycosides and aglycones (emodin, rhein, aloe-emodin, chrysophanol, physcion), stilbene glycosides (rhaponticin), and tannins. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated R. palmatum reference standard to confirm species identity and detect potential adulteration, substitution with common garden rhubarb (R. rhabarbarum or R. rhaponticum) — which lacks the pharmacopeial anthraquinone profile — or blending with other anthraquinone-containing botanicals.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic anthraquinone and stilbene profile of R. palmatum. The extract is applied alongside a certified R. palmatum reference standard and, where applicable, potential adulterant extracts (e.g., R. rhabarbarum, Rumex species), 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 anthraquinone aglycones and glycosides of R. palmatum. After development, the plate is evaluated under UV light at 254 nm and 366 nm — where anthraquinones display characteristic fluorescence — and may be further derivatized with potassium hydroxide solution or anisaldehyde-sulfuric acid reagent for enhanced 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.

Medicinal rhubarb (R. palmatum and related pharmacopeial species) is subject to adulteration and substitution with common garden rhubarb (R. rhabarbarum), which is morphologically similar but lacks the characteristic anthraquinone glycoside profile required for pharmacopeial compliance and the associated biological activity. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation — anchored by the characteristic anthraquinone pattern — that is more discriminating than single-marker assays and enables detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP and European Pharmacopoeia (Ph. Eur.) 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 D-ribose, a naturally occurring sugar used in supplements for heart health, energy metabolism, and recovery. Using LC-MS/MS, it confirms D-ribose content in powders, capsules, and beverages to ensure proper dosing and label compliance.

Samples are extracted in water or dilute acid and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified D-ribose standards, with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in g per 100 g or per serving. Values are compared to formulation targets and label claims to verify product consistency, detect adulteration, and ensure ingredient integrity.

This assay quantifies total alpha-lipoic acid (ALA) in supplements and raw materials using LC-MS/MS. It does not distinguish between the R- and S-enantiomers, but instead measures total ALA content to verify label claims, confirm potency, and ensure product consistency.

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

Results are reported in mg per g or per serving as total ALA. Values are compared to formulation targets and label claims to confirm dosing accuracy, detect underformulation, and verify product stability.

This assay quantifies rosavin, a key bioactive compound in Rhodiola rosea known for its adaptogenic and anti-fatigue properties. Using HPLC, it verifies rosavin content in botanical extracts and supplements to ensure proper standardization and support claims related to mood, energy, and stress modulation.

Samples are extracted using alcohol- or water-based solvents under light- and temperature-controlled conditions. The extract is analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified rosavin 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 label claims and standardization targets to confirm consistency in Rhodiola extracts and detect low-quality or adulterated material.

This test quantifies rosmarinic acid, a naturally occurring hydroxycinnamic acid ester and the primary water-soluble polyphenol found in rosemary, sage, lemon balm, and other Lamiaceae herbs, in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Rosmarinic acid is a key bioactive marker and standardization compound for these botanical extracts, valued for its antioxidant, anti-inflammatory, and 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 ethanol-water solvent system with sonication to ensure complete recovery of rosmarinic 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 330 nm, corresponding to the characteristic absorbance of the hydroxycinnamic acid chromophore, and quantification is performed against a multi-point external calibration curve prepared from a certified rosmarinic acid reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Rosmarinic acid is the primary standardization marker for lemon balm, rosemary, and related Lamiaceae extracts, and its accurate quantification is essential for verifying extract potency and label claim compliance. HPLC-UV at 330 nm provides the selectivity needed to resolve rosmarinic acid from co-present phenolic acids — including caffeic acid and salvianolic acids — in complex botanical matrices, delivering reliable potency data for both raw material qualification and finished product release testing.

This test quantifies rutin, a bioactive flavonoid glycoside of quercetin, in raw materials, powders, and finished botanical products. Using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS), the method achieves sensitive and specific detection with a reporting limit as low as 0.1 mg/kg. Accurate rutin measurement supports quality control and standardization of herbal supplements and nutraceuticals.

Samples are extracted with 70% methanol under sonication to release rutin from the matrix. The extract is filtered and injected into an LC-MS/MS system operating in Multiple Reaction Monitoring (MRM) mode, targeting rutin-specific precursor and product ions. Quantification is performed using a calibration curve constructed from certified rutin reference standards, with an isotopically labeled internal standard to correct for matrix effects. Method accuracy is verified through duplicate injections, spiked recovery tests, and quality control samples analyzed alongside each batch.

Results are reported in mg/g (raw material) or mg/serving (finished products). Testing confirms standardized potency in plant extracts, verifies label claims, and supports quality control for flavonoid-rich formulations.

This test quantifies S-acetyl-glutathione (SAG) — a chemically stable, acetylated form of reduced glutathione (GSH) designed to resist oxidation in the gastrointestinal tract and improve intracellular delivery — in dietary supplements and raw materials using High-Performance Liquid Chromatography (HPLC). Unlike standard reduced glutathione, S-acetyl-glutathione is protected from degradation prior to absorption, making accurate potency verification essential to confirm that the declared amount of this premium form is present and intact. Results are reported as a percentage or in milligrams per serving to support label claim substantiation and cGMP compliance.

A representative sample is accurately weighed and dissolved in a dilute acidic aqueous solution (e.g., 0.1% trifluoroacetic acid or dilute phosphoric acid) to stabilize the thiol-containing compound and prevent oxidation during sample preparation. 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 210–220 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified S-acetyl-glutathione reference standard. To distinguish SAG from reduced glutathione (GSH) and oxidized glutathione (GSSG), chromatographic resolution of these species is confirmed during method validation. Quality control standards and system suitability checks are run concurrently to confirm method accuracy and precision throughout the analytical run.

S-acetyl-glutathione is a premium, higher-cost form of glutathione that commands a price premium over standard reduced glutathione, making it a target for substitution with the less expensive unmodified form. HPLC provides the chromatographic resolution needed to specifically quantify SAG and distinguish it from GSH, GSSG, and other related compounds that may be present in glutathione-containing formulations. Accurate potency testing is essential for label claim compliance, raw material qualification, and consumer transparency, particularly given the growing market for bioavailable glutathione forms in the antioxidant supplement category.

This assay quantifies S-adenosyl-L-methionine (SAMe), a bioactive methyl donor involved in neurotransmitter synthesis, detoxification, and joint support. Using LC-MS/MS, it verifies SAMe content in supplements and functional formulas to ensure label accuracy and product stability.

Samples are extracted under acid-stabilized, light-protected conditions to prevent degradation. The extract is analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified SAMe 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 declared label claims. Testing confirms bioactive content and helps detect instability due to SAMe’s known sensitivity to heat, moisture, and pH.

This test quantifies safranal (2,6,6-trimethylcyclohexa-1,3-diene-1-carbaldehyde) — the principal volatile aldehyde responsible for saffron's characteristic aroma and a key bioactive constituent of saffron (Crocus sativus L.) stigma — in saffron raw materials, extracts, and dietary supplements using UV-Visible (UV-Vis) Spectrophotometry. Safranal, along with crocins and picrocrocin, is one of the three primary quality marker compounds defined in the ISO 3632 standard for saffron quality grading. It is increasingly recognized for its neuroprotective, antidepressant, and anxiolytic properties. UV-Vis spectrophotometry at the characteristic safranal absorption maximum provides a rapid, standardized method for safranal quantification consistent with ISO 3632 quality grading requirements. Results are reported as absorbance at 330 nm (E¹%₁cm value) or as a percentage of safranal content relative to a certified reference standard.

A representative sample is accurately weighed and extracted using an appropriate aqueous solvent (e.g., water or aqueous methanol) under controlled conditions to ensure complete extraction of the water-soluble saffron constituents. The extract is filtered and its absorbance is measured spectrophotometrically at 330 nm — the characteristic absorption maximum of safranal — against a solvent blank using a calibrated UV-Vis spectrophotometer. Safranal content is calculated using the Beer-Lambert law with the specific absorptivity value established for safranal, or by comparison to a multi-point calibration curve prepared from a certified safranal reference standard. Measurements are performed in triplicate and averaged for final quantification. Where a full saffron quality profile is required, absorbance measurements at 257 nm (picrocrocin) and 440 nm (crocins) are performed concurrently in accordance with ISO 3632.

UV-Vis spectrophotometry at 330 nm is the internationally standardized method for safranal quantification in saffron, as specified in ISO 3632 (Saffron — Specification and Test Methods), which is the globally recognized quality standard for saffron grading. The method exploits safranal's characteristic UV absorption to provide a rapid, reproducible, and cost-effective measure of this key aroma compound without the need for chromatographic separation, making it well suited to routine quality control and supplier qualification of saffron raw materials. This approach supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies safranal, a volatile compound and key bioactive in saffron (Crocus sativus) linked to mood and neurological support. Using HPLC, it measures safranal content in saffron extracts and supplements to verify label claims and ensure consistent dosing in mood, vision, and cognitive health products.

Samples are extracted using alcohol-based solvents under light-protected conditions, then analyzed by HPLC with UV detection at a safranal-specific wavelength. Quantification is performed using certified safranal standards, with internal standard correction and duplicate runs for precision.

Results are reported in mg per g or per serving. Values are compared to formulation targets and standardization benchmarks to confirm potency and detect variability due to degradation or poor-quality raw material.

This test quantifies salicin (2-(hydroxymethyl)phenyl β-D-glucopyranoside) — the principal phenolic glycoside and primary standardization marker of white willow bark (Salix alba L. and related Salix species) — in raw materials, bark extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Salicin is the naturally occurring precursor to salicylic acid, the active metabolite responsible for the analgesic and anti-inflammatory activity historically associated with willow bark. White willow bark extracts standardized to salicin content are widely used in dietary supplements for joint health, pain management support, and healthy inflammatory response. Accurate potency verification is essential for label claim substantiation and for confirming compliance with pharmacopeial and industry specifications for salicin content. 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-organic solvent system (e.g., aqueous methanol or ethanol) to ensure complete extraction of salicin and related phenolic glycosides. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 270 nm, the characteristic absorption maximum of salicin's phenolic chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified salicin reference standard. Where applicable, related phenolic glycosides — including salicortin, tremulacin, and fragilin — may be monitored simultaneously to provide a broader phenolic glycoside profile characteristic of the Salix species. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Salicin's phenolic chromophore provides strong UV absorption at 270 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 salicin from structurally related phenolic glycosides and matrix components present in willow bark extracts, ensuring that potency results reflect salicin content specifically. This specificity is important for compliance with pharmacopeial specifications — salicin content is defined in the European Pharmacopoeia (Ph. Eur.) monograph for willow bark — and for label claim accuracy in the dietary supplement market. The method supports raw material qualification, finished product release testing, and cGMP compliance under 21 CFR 111.

This test simultaneously identifies and quantifies five key naturally occurring salicylate compounds — salicin, salicylic acid, salicylaldehyde, methyl salicylate, and salicyl alcohol — in botanical raw materials, extracts, and dietary supplements using a combined HPLC and LC-MS/MS approach. Salicylates are a structurally related family of phenolic compounds found across a wide range of botanicals, most notably white willow bark (Salix alba and related Salix species), meadowsweet (Filipendula ulmaria), wintergreen (Gaultheria procumbens), and poplar (Populus spp.). Each compound plays a distinct role: salicin is the primary glycoside precursor to salicylic acid and the principal standardization marker for willow bark; salicylic acid is the pharmacologically active metabolite; salicylaldehyde and salicyl alcohol are related phenolic metabolites; and methyl salicylate is the characteristic volatile ester responsible for the distinctive wintergreen aroma. Comprehensive salicylate profiling supports potency verification, botanical authentication, and detection of adulteration or inter-species substitution. Results are reported in milligrams per gram or as a percentage for each individual compound.

A representative sample is accurately weighed and extracted using an appropriate aqueous-organic solvent system (e.g., aqueous methanol or ethanol) to ensure complete extraction of the full salicylate compound series. Isotopically labeled internal standards are added prior to extraction to correct for matrix effects and recovery variability across the structurally diverse analyte panel. The extract is filtered and analyzed by reversed-phase HPLC with UV detection at 270 nm for primary quantification of salicin and salicylic acid, which possess suitable UV chromophores at this wavelength. Salicylaldehyde, methyl salicylate, and salicyl alcohol — which have lower UV absorptivity or require greater specificity in complex matrices — are quantified by LC-MS/MS using electrospray ionization (ESI) in negative ion mode with multiple reaction monitoring (MRM) transitions selected for each target compound. Quantification of all five compounds is performed against multi-point external calibration curves prepared from certified reference standards for each individual salicylate. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

The salicylate compounds in this panel span a range of polarities, volatilities, and UV absorptivities that cannot be optimally addressed by a single analytical technique. The combined HPLC/LC-MS/MS approach leverages UV detection for the strongly absorbing, higher-concentration compounds (salicin, salicylic acid) while applying the superior sensitivity and compound-specific selectivity of MRM-based MS/MS detection for the lower-abundance and structurally less UV-active species (salicylaldehyde, methyl salicylate, salicyl alcohol). Comprehensive salicylate profiling provides a more complete characterization of the botanical's bioactive content than single-marker salicin assays, enabling authentication of botanical origin — as the relative salicylate profile is characteristic of specific plant species — and detection of adulteration or blending. This supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies salidroside, a key adaptogenic glycoside found in Rhodiola rosea. Using HPLC, it measures salidroside content in botanical extracts and supplements to verify standardization and support health claims related to mood, focus, and fatigue reduction.

Samples are extracted using water or alcohol-based solvents, then analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified salidroside 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 consistent potency and detect variability in raw material or extract quality.

This test quantifies total saponin content in botanical raw materials, extracts, and dietary supplements using UV-Visible (UV-Vis) Spectrophotometry. Saponins are a structurally diverse class of naturally occurring amphiphilic glycosides composed of a steroidal or triterpenoid aglycone (sapogenin) linked to one or more sugar chains, found in a wide range of botanicals including ginseng (Panax spp.), tribulus (Tribulus terrestris), fenugreek (Trigonella foenum-graecum), quillaja (Quillaja saponaria), and soapwort (Saponaria officinalis). Saponin content is a key standardization marker for many botanical extracts used in dietary supplements for their adaptogenic, performance-enhancing, and immune-modulating properties. UV-Vis spectrophotometry following colorimetric derivatization provides a rapid, practical method for total saponin quantification suitable for raw material screening and quality control. Results are reported as a percentage or in milligrams per gram, typically expressed as saponin equivalents relative to a reference standard.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., aqueous methanol or ethanol) to ensure complete extraction of the saponin fraction. The extract is subjected to a colorimetric derivatization reaction — most commonly the vanillin-sulfuric acid method or the Liebermann-Burchard reaction — which reacts with the sapogenin aglycone to produce a colored chromophore with characteristic absorption in the visible range (typically 540–560 nm for vanillin-sulfuric acid). Absorbance is measured spectrophotometrically against a solvent blank, and total saponin concentration is calculated by comparison to a multi-point calibration curve prepared from a certified saponin reference standard (e.g., oleanolic acid, diosgenin, or a purified saponin appropriate to the botanical source). All measurements are performed in triplicate and averaged for final quantification.

Total saponin quantification by UV-Vis colorimetry is the standard quality control approach for saponin-standardized botanical extracts, providing a rapid and accessible measure of total saponin content that is well suited to routine raw material screening and supplier qualification. While UV-Vis does not resolve individual saponin species or distinguish between steroidal and triterpenoid saponins, it is appropriate for standardized extracts where the saponin class is well characterized and total saponin content is the primary specification parameter. This method supports raw material potency verification, label claim substantiation, and cGMP compliance under 21 CFR 111, with HPLC or LC-MS/MS available as complementary methods when individual saponin profiling is required.

This assay quantifies bioactive lignans in Schisandra chinensis, primarily schisandrins and gomisins, using LC-MS/MS. These compounds are responsible for Schisandra’s adaptogenic, hepatoprotective, and antioxidant effects, and are commonly standardized in premium botanical extracts.

Samples are extracted using alcohol- or methanol-based solvents and analyzed by LC-MS/MS with compound-specific mass transitions. Quantification is performed using certified standards for schisandrin A, schisandrin B, and gomisin A, with internal standard correction and duplicate injections for accuracy.

Results are reported in mg per g or per serving for individual or total lignans. Values are compared to formulation targets and label claims to ensure potency, consistency, and identity of Schisandra extracts.

This assay quantifies sennosides A and B, the primary active glycosides found in senna leaf and pod extracts. Using HPLC, it verifies sennoside content in herbal laxative supplements to ensure proper dosing, regulatory compliance, and consistent formulation strength.

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

Results are reported in mg per g or per serving for individual sennosides and total sennosides. Values are compared to formulation targets and label claims to confirm therapeutic efficacy and prevent overdosing in stimulant laxative products.

This test quantifies the proteolytic activity of serratiopeptidase (serrapeptase) — a serine protease originally derived from Serratia marcescens bacteria and widely used in dietary supplements for its anti-inflammatory, fibrinolytic, and mucolytic properties — in raw materials and finished products, expressed in Serratiopeptidase Plasminogen Units (SPU) using a standardized activity assay. Because serratiopeptidase is an enzyme whose biological effect depends on its functional activity rather than its mass, potency is measured in activity units rather than milligrams. Accurate activity quantification is essential for label claim verification and for confirming that the declared SPU count reflects true enzymatic potency in the finished product. Results are reported in SPU per gram or per serving.

A representative sample is dissolved in a suitable aqueous buffer at a defined pH and temperature. Proteolytic activity is measured using a standardized chromogenic or caseinolytic substrate assay under controlled conditions. In the SPU method, the rate of substrate hydrolysis is measured spectrophotometrically at the appropriate wavelength, and the activity is calculated by comparison to a certified serratiopeptidase reference standard with a known SPU activity value. Assay parameters including pH, temperature, substrate concentration, and incubation time are tightly controlled, and positive and negative controls are run concurrently to confirm assay validity. Results are expressed as SPU per gram of sample.

Serratiopeptidase is an activity-dependent enzyme ingredient whose potency can be significantly affected by processing conditions, pH exposure, temperature, and formulation excipients — all of which can reduce enzymatic activity without altering total protein mass. Activity-based testing using the SPU method directly measures the functional proteolytic capacity of the enzyme, providing a physiologically meaningful measure of potency that mass-based methods cannot capture. This approach is the industry standard for serratiopeptidase specification and label claim substantiation, and supports cGMP compliance under 21 CFR 111.

This test confirms the identity of shilajit — a mineral-rich exudate from Himalayan and Altai mountain rock formations — in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic fulvic acid and dibenzo-alpha-pyrone (DBP) compound fingerprint of the sample is compared against a certified shilajit reference standard to confirm authenticity and detect adulteration with synthetic fulvic acid, humic acid, or other substitutes. Results are reported as confirmed identity or non-conforming.

A representative sample is dissolved in methanol or water and applied alongside a certified shilajit reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system and examined under UV light (254 nm and 366 nm) and after derivatization with an appropriate 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.

Shilajit is a high-value ingredient with a significant risk of adulteration, particularly with synthetic fulvic acid or humic acid preparations that mimic its appearance but lack its full bioactive compound profile. HPTLC identity testing provides a practical and defensible method for confirming authenticity, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This test quantifies silybin (also known as silibinin), the most pharmacologically active flavonolignan in silymarin complex derived from milk thistle (Silybum marianum), in dietary supplements and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Silybin exists as two diastereomers — silybin A and silybin B — and is the primary bioactive marker responsible for the hepatoprotective properties of milk thistle extract. LC-MS/MS enables specific quantification of silybin at the trace and low-dose levels present in standardized extracts, distinguishing it from the other silymarin flavonolignans including silychristin, silydianin, and isosilybin. 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 silybin from the botanical matrix. The clarified extract is injected onto a reversed-phase C18 HPLC column coupled to a triple quadrupole mass spectrometer operating in negative or positive ionization Multiple Reaction Monitoring (MRM) mode. Specific precursor-to-product ion transitions characteristic of silybin A and silybin B are monitored individually for quantification and identity confirmation, with results reported as total silybin or as individual diastereomers as required. Quantification is performed against a multi-point calibration curve prepared from certified silybin reference standards, with system suitability and QC samples run concurrently to confirm method performance.

Silybin is the primary bioactive and highest-value component of silymarin, and its accurate quantification is essential for verifying the potency of milk thistle extracts beyond total silymarin content. LC-MS/MS provides the molecular selectivity needed to distinguish silybin from its structural analogs — isosilybin, silychristin, and silydianin — which co-elute or partially overlap under HPLC-UV conditions, delivering a more precise and defensible measure of extract quality for premium milk thistle formulations.

This assay measures silymarin, a flavonolignan complex, using High-Performance Liquid Chromatography (HPLC). Analysis provides accurate quantification of total silymarin content for quality control and standardization purposes.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. Individual silymarin components are separated and detected via UV or diode-array detection, then summed to determine total silymarin content. 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.

A comprehensive nutritional profile assay that consolidates the measurement of primary macronutrients—protein, carbohydrates, and fats—and calculates the total caloric content for nutritional labeling and quality control.

The assay integrates multiple analytical methods: protein is measured using the Dumas combustion method; fat is determined by solvent extraction (e.g., Soxhlet) and gravimetric analysis; carbohydrates are calculated by difference (or summing measured sugars/starches); and caloric content is derived either via bomb calorimetry or standard conversion factors. Calibration with certified standards and duplicate analyses ensure reliability across components.

Results are reported as grams per 100 g (or per serving) for protein, carbohydrates, and fats, along with total calories per 100 g (or per serving). Consistency in these values confirms formulation accuracy, while any deviations may indicate processing or measurement issues that require further investigation.

This assay quantifies sodium beta-hydroxybutyrate (Na-BHB), a ketone salt used in ketogenic supplements for energy and metabolic support. Using HPLC, it verifies BHB content in powders and capsules to ensure proper dosing and label accuracy in performance and weight management products.

Samples are extracted in aqueous solution and analyzed by HPLC with UV or refractive index detection. Quantification is performed using certified BHB standards, with internal standard correction and duplicate injections to ensure consistent and accurate results.

Results are reported in mg per g or per serving. Values are assessed against formulation targets and declared label claims to confirm active content and detect formulation inconsistencies.

This test quantifies sodium hyaluronate — the sodium salt form of hyaluronic acid (HA), a naturally occurring glycosaminoglycan found in skin, synovial fluid, and connective tissue — in dietary supplements, topical products, and raw materials using UV-Visible (UV-Vis) Spectrophotometry. Sodium hyaluronate is widely used in beauty, joint health, and eye health formulations for its exceptional water-binding capacity and lubricating properties. UV-Vis quantification using a colorimetric assay provides a reliable and cost-effective method for confirming the declared potency of sodium hyaluronate in raw materials and finished products. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved in ultrapure water or a suitable aqueous buffer. The hyaluronic acid content is quantified using a validated colorimetric assay — typically the carbazole-sulfuric acid method or a uronic acid colorimetric assay — which reacts with the glucuronic acid residues of the hyaluronate polymer to produce a colored complex. Absorbance is measured at the appropriate wavelength (typically 530 nm for carbazole or 520 nm for similar uronic acid assays) against a reagent blank. Concentration is calculated from a multi-point calibration curve prepared from a certified sodium hyaluronate or glucuronic acid reference standard. Measurements are performed in triplicate and averaged for final quantification.

Sodium hyaluronate is a premium ingredient with significant price variation based on molecular weight and purity, making accurate potency verification important for both label claim substantiation and raw material qualification. UV-Vis colorimetric quantification targeting the uronic acid backbone of hyaluronate provides a practical and widely used method for confirming total hyaluronate content in high-purity raw materials and finished products. This method supports efficient quality control workflows and supplier qualification under 21 CFR 111.

This assay quantifies sorbitol, a widely used sugar alcohol and low-calorie sweetener, in food, beverage, and supplement products. Using HPLC, it verifies sorbitol levels to support “sugar-free,” “no added sugar,” or diabetic-friendly claims, while also ensuring formulation consistency and label accuracy.

Samples are extracted in water or dilute acid, filtered, and analyzed by HPLC with refractive index or UV detection depending on the matrix. Calibration with high-purity sorbitol standards, internal standard correction, and duplicate injections ensure reliable quantification.

Results are reported in g/100 g, g/100 mL, or per serving. The values are assessed against nutritional label claims and formulation targets. Testing confirms proper dosing of sugar alcohols and ensures that products meet regulatory definitions for “sugar-free” or “low glycemic” positioning.

This assay quantifies spermidine, a naturally occurring polyamine involved in cellular growth and longevity pathways. Using LC-MS/MS, it verifies spermidine content in dietary supplements, functional foods, and longevity formulations to confirm label accuracy and support anti-aging claims.

Samples are extracted in acidified aqueous solution and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed with certified spermidine standards, 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 declared label claims to ensure proper dosing and detect degradation or inconsistencies in raw material quality.

This test quantifies spermidine trihydrochloride, the salt form of spermidine used in dietary supplement formulations, in supplements and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Spermidine is a naturally occurring polyamine found in wheat germ, aged cheese, and other foods, and has attracted significant research interest for its role in inducing autophagy — the cellular self-cleaning process associated with healthy aging and longevity. LC-MS/MS is required for this analysis due to the highly polar, low-UV-absorbing nature of spermidine and its presence at microgram-level doses in supplement formulations. Results are reported in mg per serving or mg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using an aqueous acidic solvent or dilute hydrochloric acid solution to ensure complete solubilization of spermidine trihydrochloride. The clarified extract is injected onto a reversed-phase C18 or HILIC column coupled to a triple quadrupole mass spectrometer operating in positive ionization Multiple Reaction Monitoring (MRM) mode. Specific precursor-to-product ion transitions characteristic of spermidine are monitored for quantification and identity confirmation. Quantification is performed against a multi-point calibration curve prepared from a certified spermidine reference standard, with a stable isotope-labeled internal standard used to correct for matrix effects and ensure accurate recovery across the sample types tested.

Spermidine is a highly polar, low-molecular-weight polyamine that lacks meaningful UV absorbance, making HPLC-UV methods impractical for its direct quantification at the microgram doses used in supplement formulations. LC-MS/MS in MRM mode provides the sensitivity and molecular specificity required to accurately detect and quantify spermidine in complex matrices, distinguishing it from structurally related polyamines such as putrescine and spermine that may be co-present in wheat germ and other botanical raw materials.

This test confirms the botanical identity of spinach raw materials — including dried leaf powder, juice powder, concentrates, and extracts — using High-Performance Thin-Layer Chromatography (HPTLC). Spinach is derived from Spinacia oleracea L. (Amaranthaceae), an edible leafy plant used in dietary supplements and functional foods as a source of naturally occurring vitamins, minerals, carotenoids, chlorophylls, nitrate, and polyphenolic constituents. HPTLC identity testing produces a characteristic multi-component chromatographic fingerprint from the material’s phenolic and pigment-associated constituents. The sample fingerprint is evaluated against an authenticated S. oleracea botanical reference material and, when appropriate, reference markers representative of its phenolic profile, such as flavonoid glycosides and phenolic-acid derivatives. This comparative approach supports verification of the declared botanical source and assists in detecting substitution or dilution with other green leafy plant materials. Identity testing is qualitative; it does not establish nutrient potency, nitrate concentration, heavy-metal content, pesticide compliance, or microbiological quality.

A representative sample is accurately weighed and extracted using a validated hydroalcoholic solvent system, such as aqueous methanol or aqueous ethanol, selected to recover characteristic polar and moderately polar constituents from spinach leaf material. Where necessary, sample preparation is adapted for high-pigment, high-sugar, or high-mineral matrices to reduce chromatographic interference. The sample extract is applied alongside authenticated Spinacia oleracea reference material and applicable phytochemical reference standards onto a silica gel 60 F₂₅₄ HPTLC plate using an automated applicator. The plate is developed in a validated mobile-phase system optimized to resolve characteristic spinach phenolic and flavonoid zones. Following development and drying, chromatograms are documented under UV light at 254 nm and 366 nm and, where appropriate, after derivatization with a suitable visualization reagent, such as Natural Products Reagent/polyethylene glycol or anisaldehyde-sulfuric acid. The sample’s retention-factor values, zone colors, fluorescence, and overall fingerprint pattern are compared with the authenticated reference profile. Identity is confirmed when the characteristic sample fingerprint is concordant with that of the reference material. System suitability and concurrent quality-control samples are included with each analytical run.

HPTLC is well suited to botanical identity confirmation because it evaluates the overall chemical fingerprint of the ingredient rather than relying on a single constituent that may occur in unrelated plant species. This is particularly important for spinach-derived powders and extracts, which can vary substantially in color, nutrient composition, and marker intensity based on cultivar, growing conditions, harvest stage, and processing. Comparison against authenticated S. oleracea reference material enables assessment of the collective pattern of characteristic constituents and provides stronger identity evidence than visual inspection or a single-marker assay alone. The method supports incoming raw-material qualification and identity verification under dietary supplement cGMP requirements in 21 CFR 111.75, with HPTLC used as a fit-for-purpose chromatographic approach consistent with USP General Chapter <203>.

This test confirms the identity of spirulina — primarily Arthrospira platensis and Arthrospira maxima, cyanobacteria (blue-green microalgae) widely marketed under the common name spirulina — in raw materials, dried powders, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Spirulina is one of the most widely consumed microalgae-based dietary supplements globally, valued for its high protein content, complete amino acid profile, and rich concentration of bioactive pigments including phycocyanin (blue), chlorophylls a and b (green), and carotenoids (β-carotene, zeaxanthin). HPTLC identity testing generates a characteristic pigment-based chromatographic fingerprint that is compared against an authenticated spirulina reference standard to confirm species identity and detect potential adulteration, substitution with other microalgae (e.g., Chlorella spp.), or blending with non-algal plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or acetone) to capture the characteristic pigment profile of spirulina, including chlorophylls, carotenoids, and phycocyanin-derived chromophores. The extract is applied alongside a certified spirulina reference standard and, where applicable, potential adulterant extracts (e.g., Chlorella powder), 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 spirulina, including chlorophyll a, chlorophyll b, β-carotene, and zeaxanthin. After development, the plate is evaluated under white light and UV light at 254 nm and 366 nm, where the characteristic green, yellow, and orange pigment bands of spirulina are visualized without derivatization. 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.

Spirulina is subject to adulteration and species substitution, including blending with Chlorella or other microalgae, synthetic colorants, or non-algal plant powders, particularly given its premium market positioning and the difficulty of visual inspection of dried powder materials. HPTLC pigment fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that exploits the distinctive and characteristic pigment composition of Arthrospira species — notably the presence of phycocyanin-derived chromophores and the specific chlorophyll and carotenoid profile — to distinguish authentic spirulina from potential substitutes and adulterants. 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 measures major stevia glycosides—including stevioside and rebaudiosides A, B, C, and M—using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high specificity and sensitivity for accurate profiling of stevia sweetener composition.

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

Testing verifies sweetener composition, supports label claims, and ensures batch-to-batch consistency for stevia-based formulations.

This assay quantifies strontium, a naturally occurring trace element sometimes used in bone health supplements. Using ICP-MS, it measures strontium levels in raw materials and finished products to verify potency, confirm label claims, or monitor for excess levels as part of a heavy metal screen.

This assay quantifies strontium, a naturally occurring trace element sometimes used in bone health supplements. Using ICP-MS, it measures strontium levels in raw materials and finished products to verify potency, confirm label claims, or monitor for excess levels as part of a heavy metal screen.

Results are reported in ppm (mg/kg), µg/g, or per serving depending on the matrix. Values are compared with label claims (if strontium is an active ingredient) or safety thresholds to ensure product integrity and compliance.

This assay quantifies key dietary sugars—glucose, fructose, and sucrose—in food, beverage, and nutrition products. Using LC-MS/MS, it verifies total and individual sugar content to support nutrition label compliance and detect the presence of undeclared or added sugars in “no sugar” or “natural” claims.

Samples are extracted in water or dilute acid, filtered, and injected into the LC-MS/MS system. Detection is performed using mass-specific transitions for each sugar. Quantification is achieved using certified sugar standards, internal standard correction, and duplicate injections to ensure precise and defensible results.

Results are reported in g/100 g, g/100 mL, or per serving. Values are assessed against label claims and regulatory thresholds (e.g., FDA nutrition labeling). This panel helps validate “no added sugar” or “low sugar” claims and ensures brand transparency in functional beverages, juices, and powdered mixes.

This test quantifies sulforaphane, the bioactive isothiocyanate derived from enzymatic hydrolysis of glucoraphanin in broccoli (Brassica oleracea) sprout and seed extracts, in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Sulforaphane is the primary bioactive compound responsible for the antioxidant, anti-inflammatory, and phase II enzyme-inducing properties of broccoli-derived ingredients, and its direct quantification confirms the amount of active compound present rather than its precursor. Results are reported in mg per serving or µmol per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using an acetonitrile-water or methanol-water solvent system with sonication to ensure complete recovery of sulforaphane from the matrix. Where the sample contains intact myrosinase enzyme, a controlled enzymatic hydrolysis step may be performed prior to extraction to convert residual glucoraphanin to sulforaphane before analysis. The clarified extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 210–254 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified sulforaphane reference standard. All sample preparation steps are conducted promptly to minimize sulforaphane degradation, and system suitability and QC samples are run concurrently to confirm method performance.

While glucoraphanin quantification measures the precursor pool, direct sulforaphane measurement confirms the actual bioactive content present in the finished product — a distinction that matters for products where myrosinase is co-formulated or where conversion has already occurred during processing. HPLC-UV provides adequate sensitivity and selectivity for sulforaphane quantification in most broccoli extract matrices, and its use alongside or in place of glucoraphanin testing gives brands a more complete picture of product potency for label claim substantiation and consumer transparency.

This test confirms the identity and source authenticity of sunflower lecithin — a complex mixture of phospholipids, glycolipids, and neutral lipids derived from sunflower (Helianthus annuus L.) seeds — in raw materials and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Sunflower lecithin is increasingly used as a soy-free and non-GMO alternative to soy lecithin in dietary supplements and functional foods, valued for its phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylinositol (PI) content. HPTLC identity testing generates a characteristic phospholipid class fingerprint that is compared against an authenticated sunflower lecithin reference standard to confirm source identity and detect potential substitution with soy lecithin or other lecithin sources, which is of particular importance for allergen management and non-GMO labeling claims.

A representative sample is accurately weighed and dissolved in an appropriate lipid solvent system (e.g., chloroform/methanol or dichloromethane/methanol) to ensure complete dissolution of the phospholipid fraction. The extract is applied alongside a certified sunflower lecithin reference standard and, where applicable, soy lecithin and other potential substitute lecithin sources, 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 major phospholipid classes characteristic of lecithin — including phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and lysophosphatidylcholine. After development, the plate is derivatized with an appropriate reagent (e.g., molybdenum blue reagent for phospholipid-specific visualization, or primuline under UV 366 nm) and evaluated under white light and UV light at 254 nm and 366 nm. The resulting phospholipid class fingerprint is compared visually and, where applicable, by densitometric analysis to the authenticated sunflower lecithin reference standard.

Sunflower lecithin and soy lecithin share similar phospholipid class compositions, making source authentication by simple phospholipid class profiling alone insufficient for definitive identity confirmation. HPTLC fingerprinting of the full lipid class profile — including characteristic differences in the relative proportions and minor lipid constituents between sunflower and soy lecithin — provides a practical and discriminating identity confirmation method for routine quality control. This is particularly important for products marketed as soy-free, allergen-free, or non-GMO, where substitution of sunflower lecithin with soy lecithin would constitute both a labeling violation and a potential allergen risk. The method supports raw material qualification, allergen management, and cGMP compliance under 21 CFR 111.

This assay quantifies p-synephrine, the primary active alkaloid found in bitter orange (Citrus aurantium). Using HPLC, it verifies synephrine content in thermogenic, pre-workout, and weight management supplements to confirm label accuracy and support safe, standardized dosing.

Samples are extracted using aqueous or alcohol-based solvents and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified p-synephrine 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 declared label claims and regulatory thresholds to ensure dosing accuracy and detect possible adulteration or mislabeling.

This test quantifies tartaric acid — a naturally occurring organic acid found in grapes, tamarinds, and certain botanical ingredients, and widely used as an acidulant, stabilizer, and excipient in food and dietary supplement formulations — using High-Performance Liquid Chromatography (HPLC). Accurate measurement of tartaric acid is important for verifying ingredient identity and concentration, ensuring formulation consistency, and confirming compliance with permitted use levels in finished products. Results are reported as a percentage or in milligrams per gram to support label claim substantiation and cGMP compliance.

A representative sample is accurately weighed and dissolved or extracted in ultrapure water or a dilute mobile phase-compatible solvent, then filtered through a 0.2 µm membrane prior to injection. Separation is performed by reversed-phase HPLC or ion-exclusion HPLC using a suitable column, with UV detection at approximately 210 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified tartaric acid reference standard. System suitability is confirmed prior to sample analysis, and quality control standards are run at regular intervals throughout the analytical sequence to ensure accuracy and precision.

Tartaric acid is used across a broad range of food and supplement applications as an acidulant, chelating agent, and excipient, and its concentration can directly affect product pH, stability, and taste profile. HPLC provides the specificity and accuracy needed to quantify tartaric acid in complex matrices, distinguishing it from other organic acids such as citric, malic, and oxalic acid that may co-occur in botanical or food-derived ingredients. This test supports both raw material qualification and finished product release testing in compliance with 21 CFR 111 cGMP requirements.

This assay quantifies taurine, a sulfur-containing amino acid commonly used in energy drinks, hydration products, and performance supplements. Using LC-MS/MS, it verifies taurine content to confirm label accuracy and support consistency in functional formulations.

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

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

This assay quantifies total terpene lactones (including ginkgolides A, B, C, and bilobalide) in Ginkgo biloba extracts using High-Performance Liquid Chromatography (HPLC). Terpene lactones are characteristic markers required for quality control and standardization of Ginkgo supplements.

Samples are extracted and analyzed by HPLC under validated chromatographic conditions. Individual terpene lactones are separated and quantified against certified reference standards. Results are summed to provide total terpene lactone content.

Results are reported as % w/w (raw materials) or mg/serving (finished products). Testing verifies standardized potency (often paired with flavonol glycosides for full Ginkgo profile), confirms raw material authenticity, and ensures label claim accuracy.

This assay quantifies theacrine, a purine alkaloid structurally similar to caffeine and marketed as TeaCrine. Using HPLC, it verifies theacrine content in energy, nootropic, and performance supplements to ensure consistent dosing and support claims related to mental clarity and stimulant-free endurance.

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

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm potency and detect underformulation or substitution with similar alkaloids.

This test quantifies theobromine, a methylxanthine compound commonly found in cocoa, chocolate products, and botanical extracts, using High-Performance Liquid Chromatography (HPLC). Accurate measurement of theobromine is important for quality control and regulatory compliance in raw materials, finished products, capsules, and powders. The method provides detection limits suitable for trace-level quantification, reported in mg/kg or mg/L depending on the sample matrix.

Samples are prepared by solvent extraction using methanol-water (80:20 v/v) followed by filtration to remove particulates. The extract is injected into an HPLC system equipped with a C18 reversed-phase column and UV detection set at 272 nm, specific for theobromine absorption. Quantification is achieved by comparing peak areas to a calibration curve generated from certified theobromine reference standards across a defined concentration range. Quality control includes duplicate injections, analysis of spiked samples to assess recovery, and periodic injection of quality control standards to ensure method precision and accuracy.

Results are reported in mg/g (raw material) or mg/serving (finished products). Testing verifies product standardization, confirms label claims, and ensures quality consistency in cocoa- or caffeine-related formulations.

This test measures the concentration of threonic acid (threonate), a small organic acid relevant for quality control in raw materials, finished products, capsules, and powders. The LC-MS/MS method offers precise quantification with high sensitivity and selectivity, enabling differentiation from closely related metabolites. Results are reported in micrograms per gram (µg/g) or milligrams per liter (mg/L) depending on the sample matrix.

Samples are prepared by extracting 0.5 g of powdered material or 1 mL of liquid with 5 mL of 0.1% formic acid in water, followed by centrifugation and filtration. The clear extract is injected into an LC-MS/MS system equipped with a reversed-phase column, using a gradient of water and acetonitrile both containing 0.1% formic acid. Threonic acid is detected in multiple reaction monitoring (MRM) mode with negative electrospray ionization. Quantification is performed using a calibration curve constructed from certified threonic acid standards, with an isotopically labeled internal standard added to each sample to correct for matrix effects. Method accuracy is verified through duplicate injections, quality control samples, and spike recovery experiments.

Results are reported in mg/g (raw material) or mg/serving (finished product). Testing confirms label accuracy, ensures standardized potency in mineral threonate salts (e.g., magnesium L-threonate), and supports product consistency.

This test quantifies thymoquinone (2-isopropyl-5-methylbenzo-1,4-quinone) — the principal bioactive constituent of black seed oil and black seed (Nigella sativa L.) extracts — in raw materials, fixed oils, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Thymoquinone is responsible for a significant portion of the pharmacological activity attributed to N. sativa, including its antioxidant, anti-inflammatory, immunomodulatory, and hepatoprotective effects. Accurate potency verification is essential for standardizing black seed oil and extract quality, verifying label claims, and ensuring that the declared amount of this key bioactive 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 dissolved in an appropriate organic solvent, typically methanol or acetonitrile, to ensure complete dissolution of thymoquinone. For black seed oil samples, a dilution step in an appropriate organic solvent is performed prior to injection. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 254 nm, the characteristic absorption maximum of thymoquinone's quinone chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified thymoquinone reference standard. Where applicable, related volatile constituents such as thymohydroquinone and thymol may be monitored simultaneously to provide a broader phytochemical profile. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Thymoquinone's quinone chromophore provides strong UV absorption at 254 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 thymoquinone from structurally related compounds — including thymohydroquinone, dithymoquinone, and thymol — that co-occur in N. sativa oil and extracts, ensuring that potency results reflect thymoquinone content specifically. This specificity is important for label claim accuracy and for confirming the quality of black seed raw materials, which can vary considerably in thymoquinone content depending on geographic origin, variety, and processing conditions. The method supports raw material qualification, finished product release testing, and cGMP compliance under 21 CFR 111.

This assay quantifies tianeptine, a tricyclic compound used in cognitive and mood-enhancing formulations. Using HPLC, it verifies tianeptine content in raw materials and finished supplements to confirm dosing accuracy, product safety, and compliance with regulatory thresholds in nootropic or mood-support products.

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

Results are reported in mg per g or per serving. Values are assessed against label claims and formulation specifications to ensure proper dosing, detect mislabeling, or identify potentially unsafe concentrations.

This assay confirms the identity of Tongkat Ali using High-Performance Thin-Layer Chromatography (HPTLC), a pharmacopeial method widely used for botanical authentication. It distinguishes genuine Tongkat Ali from common adulterants through unique compound fingerprinting.

Samples are extracted with appropriate solvents and applied to a silica gel plate alongside a reference standard. The plate is developed in a controlled mobile phase and visualized under UV light and post-derivatization. The sample chromatogram is compared to the reference fingerprint for botanical confirmation.

Results are reported as Pass/Fail or Match/No Match to the authenticated Tongkat Ali standard. This method helps confirm raw material authenticity and detect substitution or dilution with non-Eurycoma species.

This test quantifies total bacopa glycosides — the collective measure of bacosides and bacopasides responsible for the cognitive and neuroprotective activity of Bacopa monnieri — in botanical extracts, raw materials, and dietary supplements using UV-Visible (UV-Vis) Spectrophotometry. Bacopa extracts are commercially standardized to a declared total glycoside content, and this method provides a rapid, cost-effective means of verifying that the extract meets its standardization specification. Results are reported as a percentage of total bacopa glycosides to support label claim substantiation and incoming material qualification.

A representative sample is accurately weighed and extracted using a suitable solvent such as methanol or aqueous ethanol. The extract is reacted with a chromogenic reagent — typically vanillin-sulfuric acid or a similar colorimetric agent — under controlled conditions to produce a colored complex with the saponin glycosides present in the extract. The absorbance of the resulting solution is measured at the appropriate wavelength (typically 540–550 nm) against a reagent blank. Total glycoside content is calculated using the Beer-Lambert law with a specific extinction coefficient or by comparison to a calibration curve prepared from a bacoside reference standard. Measurements are performed in triplicate and averaged for final quantification.

Bacopa monnieri extracts are standardized on the basis of total bacoside content, and UV-Vis colorimetric quantification provides a practical and widely used method for verifying this specification in both raw materials and finished products. While it does not resolve individual glycoside species, the total glycoside measurement is the industry-standard potency parameter for Bacopa and is directly linked to the declared label claim. This method supports efficient quality control workflows and supplier qualification under 21 CFR 111.

This test determines the total concentration of biophenolic compounds in virgin olive oil using the International Olive Council (COI) high-performance liquid chromatography method. Olive-oil biophenols are a chemically diverse group of naturally occurring polar compounds that include hydroxytyrosol, tyrosol, phenolic acids, lignans, flavonoids, and secoiridoid derivatives, including compounds related to oleuropein and ligstroside. These constituents contribute to the sensory characteristics, oxidative stability, and compositional quality of virgin olive oil. The COI HPLC method measures the extractable phenolic fraction chromatographically and reports the combined result as milligrams of tyrosol equivalents per kilogram of oil (mg/kg tyrosol equivalents). This result is commonly used to characterize the phenolic composition of extra-virgin and virgin olive oils and to monitor effects of cultivar, growing conditions, milling practices, storage, and age.

A representative virgin olive-oil sample is accurately weighed and extracted with a validated methanol/water solvent system in accordance with the applicable COI method. A qualified internal standard, commonly syringic acid under the COI procedure, is added to control extraction and analytical response. The polar extract is separated from the oil phase, clarified as required, and analyzed by reversed-phase HPLC using a C18 column with UV or diode-array detection, typically monitored at the wavelength specified by the applicable COI method for phenolic determination. The chromatographic profile is evaluated for the collective set of peaks corresponding to the biophenolic fraction. Peak areas are quantified relative to the applicable calibration and internal-standard response, and the total is expressed as mg/kg tyrosol equivalents in accordance with the COI calculation procedure. Reagent blanks, system-suitability checks, calibration or response-verification standards, duplicate preparations, and concurrent quality-control samples are assessed to verify chromatographic performance, accuracy, precision, and acceptable background contribution.

Unlike non-specific colorimetric assays, COI HPLC separates the phenolic fraction from the oil matrix and measures the chromatographic contribution of individual and unresolved biophenolic constituents within the validated method scope. This provides a more compositionally relevant and defensible total-biophenol value for virgin olive oil. Expressing the result as tyrosol equivalents establishes a standardized reporting basis for comparing oils and monitoring quality over time. The assay supports supplier qualification, lot-to-lot comparison, product-development decisions, storage-stability programs, and accurate content specification for virgin olive-oil ingredients. It should be applied using the current applicable version of the COI method and only to matrix types within the method’s validated scope.

This test quantifies total capsaicinoids — primarily capsaicin and dihydrocapsaicin, along with minor analogs — in chili pepper extracts, dietary supplements, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Capsaicinoids are the bioactive alkaloids responsible for the thermogenic and metabolic effects of capsicum-based ingredients, and their total concentration is the standard potency marker used to define extract strength and standardization. Results are reported in mg per serving or as a percentage of extract weight (e.g., Scoville Heat Units equivalent or % capsaicinoids) to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using an acetonitrile or methanol-water solvent system with sonication to ensure complete recovery of capsaicin, dihydrocapsaicin, and related capsaicinoid analogs 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 280 nm, and quantification is performed against a multi-point external calibration curve prepared from certified capsaicin and dihydrocapsaicin reference standards. Total capsaicinoids are reported as the sum of all detected analogs, with system suitability and QC samples run concurrently to confirm method accuracy and reproducibility.

Capsaicinoid content is the defining quality and potency marker for capsicum-based supplement ingredients, and accurate quantification of the full capsaicinoid profile — not just capsaicin alone — is essential for verifying standardization levels and label claims. HPLC-UV at 280 nm provides the selectivity needed to resolve individual capsaicinoid analogs from one another and from co-extractives in complex botanical matrices, delivering reliable data for both raw material qualification and finished product release testing.

This test confirms the identity of Cissus quadrangularis L. (veldt grape or devil's backbone) in raw materials, stem powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Cissus quadrangularis is a succulent vine native to Africa and Asia, used extensively in Ayurvedic medicine and modern dietary supplements for its role in supporting bone fracture healing, joint health, and weight management. Its characteristic phytochemical profile includes ketosteroids (osteogenic steroids), triterpenoids (including β-sitosterol and friedelin), flavonoids (quercetin, kaempferol), stilbene derivatives (resveratrol), and ascorbic acid. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated C. quadrangularis reference standard to confirm species identity and detect potential adulteration, substitution with other Cissus 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 C. quadrangularis, including flavonoids, triterpenoids, and ketosteroid constituents. The extract is applied alongside a certified C. quadrangularis 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 C. quadrangularis. After development, the plate is derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid or Natural Products Reagent A / NP/PEG) 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.

Cissus quadrangularis raw materials are subject to adulteration and species substitution, including replacement with other Cissus species or unrelated plant materials 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 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 quantifies total curcuminoids — the collective term for the three principal bioactive diarylheptanoid pigments of turmeric (Curcuma longa L.): curcumin (the most abundant, typically 60–70%), demethoxycurcumin, and bisdemethoxycurcumin — in raw materials, turmeric extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Curcuminoids are the primary quality markers and bioactive constituents responsible for the antioxidant, anti-inflammatory, and other health-promoting properties attributed to turmeric. Total curcuminoid content is the standard potency specification for turmeric extracts, with commercial standardized extracts typically containing 95% total curcuminoids. Accurate HPLC quantification of all three individual curcuminoids — and their sum as total curcuminoids — is essential for label claim substantiation and for confirming that the full curcuminoid profile is present at the declared level. Results are reported as a percentage or in milligrams per gram or per serving for each individual curcuminoid and as a total.

A representative sample is accurately weighed and dissolved in an appropriate organic solvent, typically methanol or acetonitrile, to ensure complete dissolution of the curcuminoid fraction. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 425 nm, the characteristic absorption maximum of the curcuminoid chromophore arising from the extended conjugated diene-ketone system. Quantification of each individual curcuminoid (curcumin, demethoxycurcumin, and bisdemethoxycurcumin) is performed against a multi-point external calibration curve prepared from certified reference standards for each compound. Total curcuminoids are reported as the sum of the three individually quantified curcuminoid concentrations. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

HPLC with UV detection at 425 nm is the method of choice for total curcuminoid quantification, providing the chromatographic resolution necessary to separately identify and quantify all three curcuminoid species — curcumin, demethoxycurcumin, and bisdemethoxycurcumin — which cannot be individually resolved by UV-Vis spectrophotometry alone. Individual quantification of each curcuminoid is important because their relative proportions vary between turmeric varieties, growing regions, and extraction processes, and because each curcuminoid contributes independently to the total bioactive profile. This approach provides more complete and accurate potency data than total curcuminoid UV-Vis methods and supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay measures total free sterols—campesterol, β-sitosterol, and stigmasterol—using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The panel provides selective and sensitive quantification of individual sterols and their combined total for quality control and standardization.

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

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

This test estimates total phenolic compounds in botanical raw materials, extracts, foods, beverages, and finished dietary supplement products using UV-visible spectrophotometry. The assay is commonly based on the Folin–Ciocalteu colorimetric reaction, in which phenolic compounds and other reducing constituents react with the Folin–Ciocalteu reagent under alkaline conditions to form a blue chromophore. The intensity of the developed color is proportional to the sample’s measured reducing capacity within the validated assay range. Results are expressed as gallic acid equivalents (GAE), typically as mg GAE/g, mg GAE/serving, or % GAE, depending on the sample type and product specification. Total phenolic content is useful as a broad compositional measurement for polyphenol-rich materials, including fruit, tea, herb, spice, seed, and botanical extracts.

A representative sample is accurately weighed and extracted with a validated solvent system, commonly aqueous methanol, aqueous ethanol, or another suitable hydroalcoholic solvent selected for the product matrix. The extract is clarified by centrifugation and/or filtration and diluted into the validated analytical range. An aliquot is combined with Folin–Ciocalteu reagent and an alkaline solution, such as sodium carbonate, then incubated under controlled time and temperature conditions to allow color development. Absorbance is measured using a UV-Vis spectrophotometer, commonly at approximately 765 nm. Quantification is performed against a multi-point calibration curve prepared from a qualified gallic acid reference standard, and results are calculated and reported as gallic acid equivalents. Reagent blanks, calibration-verification standards, duplicate preparations, matrix controls or spike-recovery samples, and concurrent quality-control samples are assessed with each batch to verify linearity, accuracy, precision, and acceptable background response.

The Folin–Ciocalteu UV-Vis assay is a practical, efficient, and widely used method for monitoring the overall phenolic-associated reducing capacity of botanical materials. It is particularly valuable for comparing lots, monitoring extraction consistency, establishing ingredient specifications, and trending the effect of processing or storage on phenolic-rich materials. Unlike targeted chromatographic assays, total phenolic analysis provides a composite measurement rather than quantification of individual compounds. It therefore serves as a useful screening and quality-control tool, while targeted HPLC or LC-MS methods may be required when a specific phenolic compound or standardized constituent must be quantified. The assay supports raw-material qualification and dietary supplement cGMP quality-control documentation under 21 CFR 111.

This assay quantifies total polyphenols using the Folin-Ciocalteu colorimetric method with UV-Vis detection. The method reflects the combined reducing capacity of phenolic compounds present in the sample.

Samples are extracted and reacted with the Folin-Ciocalteu reagent under alkaline conditions, producing a blue chromophore. Absorbance is measured by UV-Vis spectrophotometry and compared against a gallic acid calibration curve to calculate total polyphenol content.

Results are reported as mg gallic acid equivalents (GAE) per g (solids) or per serving (finished products). Testing provides a standardized measure of antioxidant potential, verifies label claims, and ensures batch-to-batch consistency.

This test quantifies total releasable moringin (4-(α-L-rhamnopyranosyloxy)benzyl isothiocyanate) — the primary bioactive isothiocyanate derived from Moringa oleifera — in raw materials, standardized extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Moringin is enzymatically released from its glucosinolate precursor, glucomoringin, upon cell disruption or digestion, and is the compound responsible for much of Moringa's documented antioxidant, anti-inflammatory, and chemoprotective activity. Measuring total releasable moringin — rather than the precursor glucosinolate alone — provides a functionally meaningful measure of the extract's true bioactive potential. Results are reported as a percentage or in milligrams per gram to support label claim substantiation and cGMP compliance.

A representative sample is accurately weighed and subjected to a controlled enzymatic hydrolysis step using myrosinase (either endogenous or exogenously added) under defined conditions of pH, temperature, and incubation time to convert glucomoringin to its active moringin isothiocyanate form. Following hydrolysis, the reaction is quenched and the moringin is extracted into an appropriate organic solvent such as methanol or acetonitrile. The extract is filtered through a 0.2 µm membrane and analyzed by reversed-phase HPLC on a C18 column, with UV detection at approximately 227 nm. Quantification is performed against a multi-point external calibration curve prepared from a certified moringin or isothiocyanate reference standard. Quality control samples and system suitability checks are run concurrently to confirm method accuracy and precision.

Glucomoringin, the precursor to moringin, is biologically inactive in its intact form and must be hydrolyzed by myrosinase to release the active isothiocyanate. Measuring only total glucosinolate content therefore overestimates the functional potency of a Moringa extract if myrosinase activity has been compromised by heat, processing, or storage. Quantifying total releasable moringin — the fraction that can actually be converted to the bioactive form — provides a more accurate and physiologically relevant measure of extract quality and efficacy. HPLC with UV detection provides the specificity and sensitivity needed to quantify moringin in complex botanical matrices, supporting both supplier qualification and label claim verification under 21 CFR 111.

This assay quantifies total saponins—bioactive glycosides found in many botanicals—using UV-Visible spectrophotometry (UV-Vis). The method provides a reliable estimate of total saponin concentration across diverse plant extracts such as ginseng, ashwagandha, fenugreek, and quinoa.

Samples are extracted and reacted with colorimetric reagents that form a measurable chromophore with saponins. Absorbance is read at a defined wavelength using a UV-Vis spectrophotometer, and concentrations are determined relative to a reference standard curve.

Testing verifies standardized potency, supports label claims, and ensures consistent quality across saponin-containing botanical products.

This test measures total Vitamin A content by quantifying retinol, retinyl acetate, retinyl palmitate, alpha-carotene, and beta-carotene in raw materials, finished products, capsules, and powders. Using High-Performance Liquid Chromatography (HPLC), it provides precise levels of both preformed Vitamin A and provitamin A carotenoids, essential for verifying product potency and meeting nutritional labeling standards. Results are reported in micrograms or International Units (IU) per serving or weight.

Samples are saponified with ethanolic potassium hydroxide to release Vitamin A compounds, followed by extraction into hexane. The hexane layer is evaporated and reconstituted in mobile phase before injection. Separation is performed using HPLC with a C18 column and detection at 325 nm for retinol derivatives and 450 nm for carotenoids. Quantification is achieved by comparing peak areas to calibration curves generated from certified reference standards for each analyte. Quality control includes duplicate injections, spike recovery tests, and analysis of standard reference materials to ensure accuracy and precision.

Results are reported in µg/g or mg/g (raw materials) or per-serving values for finished products. Testing verifies total Vitamin A potency, supports label claims, and ensures product consistency.

This test measures the concentrations of Vitamin K1 (phylloquinone) and the key Vitamin K2 forms (menaquinones MK-4 and MK-7) in raw materials, finished products, capsules, and powders. Accurate quantification of these fat-soluble vitamins is essential for ensuring product quality and supporting nutritional labeling and regulatory compliance. The analysis is performed using High-Performance Liquid Chromatography (HPLC) with sensitivity suitable for detecting low microgram levels per gram of sample.

Samples are first extracted using an organic solvent mixture to isolate fat-soluble vitamers, followed by saponification to release bound forms. The extract is then injected into an HPLC system equipped with a reverse-phase column and detected using fluorescence detection at excitation and emission wavelengths optimized for Vitamin K compounds. Quantification is achieved by comparing peak areas to calibration curves prepared from certified reference standards for phylloquinone, MK-4, and MK-7. Method accuracy and precision are confirmed through duplicate sample analyses, inclusion of quality control samples, and spike recovery tests.

Results are reported in µg/g (raw materials) or µg/serving (finished products). Testing verifies Vitamin K potency, confirms label claims, and ensures batch-to-batch consistency.

This test confirms the botanical identity of Tribulus terrestris in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic steroidal saponin fingerprint of the sample is compared against a certified Tribulus terrestris reference standard to confirm species authenticity and detect substitution with other Tribulus 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 Tribulus terrestris reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, derivatized with anisaldehyde-sulfuric acid 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.

Tribulus terrestris is subject to adulteration with other Tribulus species that differ significantly in their saponin profile and potency. HPTLC identity testing provides a defensible species confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay measures tributyrin, a triglyceride form of butyric acid used to support gut health and intestinal integrity. HPLC analysis provides accurate quantification of tributyrin content in raw materials and finished products, ensuring standardized delivery of butyrate equivalents.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. Tributyrin is separated from other triglycerides and lipid components and detected using appropriate detectors (e.g., UV or ELSD). Quantitation is performed against certified reference standards with calibration curves and replicate analyses to ensure accuracy and reproducibility.

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

This assay measures trimethylamine (TMA), a volatile amine associated with spoilage and metabolic byproducts, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high sensitivity and selectivity for accurate quantification in complex matrices.

This assay measures trimethylamine (TMA), a volatile amine associated with spoilage and metabolic byproducts, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high sensitivity and selectivity for accurate quantification in complex matrices.

Testing supports quality assurance, verifies specification compliance, and helps identify spoilage or processing-related issues.

This assay quantifies trimethylglycine (betaine), a naturally occurring methyl donor used in supplements for cardiovascular, cognitive, and athletic performance support. Using LC-MS/MS, it verifies betaine content in powders, capsules, and functional beverages to ensure proper dosing and formulation accuracy.

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

Results are reported in mg per g or per serving. Values are compared with formulation targets and label claims to confirm dosing consistency and detect potential mislabeling or underformulation.

This assay quantifies tauroursodeoxycholic acid (TUDCA), a bile acid conjugate used in liver support and cellular health supplements. Using HPLC, it verifies TUDCA content in capsules, powders, and functional formulations to confirm label accuracy and ensure consistent therapeutic dosing.

Samples are extracted in aqueous or methanolic solution and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified TUDCA standards, with internal standard correction and duplicate injections to ensure accurate and reproducible results.

Results are reported in mg per g or per serving. Values are assessed against formulation targets and declared label claims to verify potency and detect degradation or underformulation.

This assay quantifies turkesterone, an ecdysteroid compound typically derived from Ajuga turkestanica. Using HPLC, it verifies turkesterone content in capsules, powders, and botanical extracts to confirm label claims and support product consistency in sports performance and adaptogenic formulations.

Samples are extracted using methanol or ethanol-based solvents under temperature-controlled conditions, then analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified turkesterone standards, with internal standard correction and duplicate injections to ensure accuracy.

Results are reported in mg per g or per serving. Values are compared to formulation targets and declared label claims to confirm active content and detect adulteration or low-potency extracts.

This test confirms the botanical identity of turmeric (Curcuma longa) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic curcuminoid fingerprint — including curcumin, demethoxycurcumin, and bisdemethoxycurcumin — of the sample is compared against a certified Curcuma longa reference standard to confirm species authenticity and detect substitution with other Curcuma species, synthetic curcumin colorants, or unrelated botanical materials. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or ethanol and applied alongside a certified Curcuma longa reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, examined under UV at 366 nm to visualize the characteristic fluorescent curcuminoid bands, and the resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and fluorescence profile.

Turmeric is one of the most widely consumed botanical ingredients globally and is a well-documented target for adulteration — including spiking with synthetic curcumin, addition of lead chromate as a yellow colorant, and substitution with other Curcuma species. HPTLC identity testing based on the curcuminoid fingerprint provides a rapid and defensible species confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay quantifies ubiquinol, the reduced and bioactive form of Coenzyme Q10, in supplements and functional products. Using HPLC, it verifies ubiquinol content to support label accuracy and ensure stability in formulations targeting mitochondrial health and oxidative stress.

Samples are extracted under oxygen- and light-protected conditions to prevent oxidation to ubiquinone. The extract is analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified ubiquinol standards, with internal standard correction and duplicate runs for precision.

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

This assay measures undecylenic acid, an unsaturated fatty acid commonly used in supplements and topical formulations. Testing ensures accurate formulation and confirms the presence and concentration of this active ingredient in raw materials and finished products.

Samples are analyzed under validated laboratory conditions appropriate for fatty acid quantification. Certified reference standards and quality controls are used to ensure accurate and reproducible results.

Testing verifies label claims, confirms ingredient purity, and supports batch-to-batch consistency.

This assay quantifies uridine-5'-monophosphate (UMP), a nucleotide used in nootropic and neurological support supplements. Using HPLC, it verifies UMP content in capsules, powders, and functional formulations to ensure accurate dosing and support claims related to brain health and cognitive performance.

Samples are extracted using water or buffered aqueous solvents and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified UMP 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 formulation targets and label claims to confirm potency, standardization, and detect degradation over time.

This assay quantifies urolithin A, a gut-derived metabolite of ellagitannins known for its role in promoting mitophagy and cellular energy. Using HPLC, it verifies urolithin A content in longevity and performance supplements to confirm label accuracy and consistent bioactive delivery.

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

Results are reported in mg per g or per serving. Values are assessed against formulation targets and label claims to confirm standardized dosing and detect degradation or variability in raw material quality.

This test quantifies ursolic acid, a naturally occurring pentacyclic triterpenoid found in the waxy coatings of many fruits, herbs, and botanical extracts including apple peel, rosemary, and holy basil (Ocimum sanctum), in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Ursolic acid has been studied for its role in muscle protein synthesis, anti-inflammatory activity, and metabolic regulation, and serves as a key potency and standardization marker for relevant botanical extracts. 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 or reflux to ensure complete recovery of ursolic 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 ursolic acid reference standard. Careful mobile phase optimization and baseline management are applied to achieve adequate sensitivity at 210 nm, and system suitability and QC samples are run concurrently to confirm method accuracy and reproducibility.

Ursolic acid is structurally similar to oleanolic acid and other pentacyclic triterpenoids that are commonly co-present in botanical extracts, requiring a validated HPLC method to resolve and accurately quantify it as a distinct compound. Detection at 210 nm demands rigorous method control to minimize baseline interference from solvent and matrix components, making QC oversight particularly important for reliable and reproducible results across raw material and finished product testing.

This assay quantifies valerenic acid, a key bioactive compound in valerian root (Valeriana officinalis) associated with calming and sedative effects. Using LC-MS/MS, it verifies valerenic acid content in herbal extracts and sleep-support supplements to confirm potency and standardization.

Samples are extracted using alcohol-based solvents and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified valerenic acid 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 ensure proper dosing and detect degradation or variability in botanical quality.

This assay quantifies vanadium, a trace element that may occur naturally in soil, water, or raw materials. Using ICP-MS, it detects and measures vanadium content in supplements, food products, and ingredients to confirm compliance with quality specifications and safety guidelines.

Samples are digested using acid-based microwave or wet digestion protocols and analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS). Quantification is performed using certified vanadium standards, with internal standard correction and duplicate injections to ensure reproducibility and precision.

Results are reported in ppm (mg/kg) or µg per serving. Values are compared to formulation specifications or international safety thresholds (e.g., USP, FDA, EFSA) to verify compliance and identify potential contamination risks.

This test quantifies vinpocetine (ethyl apovincaminate) — a semi-synthetic alkaloid derivative of vincamine, itself a natural alkaloid isolated from the lesser periwinkle (Vinca minor L.) — in raw materials and dietary supplements using High-Performance Liquid Chromatography (HPLC). Vinpocetine is used in dietary supplements and pharmaceutical preparations for its purported role in supporting cerebral circulation, cognitive function, and neuroprotection. It is subject to ongoing regulatory scrutiny in the United States, where the FDA has raised questions regarding its status as a lawful dietary ingredient under 21 CFR 101 and 21 CFR 111. Accurate potency verification is essential for label claim substantiation and for confirming that the declared amount of vinpocetine 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 dissolved in an appropriate organic solvent, typically methanol or acetonitrile, to ensure complete dissolution of vinpocetine. The extract is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 268 nm, the characteristic absorption maximum of vinpocetine's indole chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified vinpocetine reference standard. Where applicable, related alkaloids such as vincamine may be monitored simultaneously to assess raw material purity and origin. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Vinpocetine's indole chromophore provides strong UV absorption at 268 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 vinpocetine from structurally related vinca alkaloids and matrix components, ensuring that potency results reflect vinpocetine content specifically. This specificity is important for label claim accuracy and for confirming the identity and purity of vinpocetine raw materials, which may vary in quality across suppliers. The method supports raw material qualification, finished product release testing, and cGMP compliance under 21 CFR 111.

This assay quantifies retinyl palmitate, a fat-soluble and commonly used form of vitamin A, in food and supplement samples. It measures vitamin A content using HPLC to verify potency and support accurate labeling in fortified formulations.

Samples are extracted under light- and oxygen-protected conditions to prevent degradation. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity retinyl palmitate standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in IU or mcg per 100 g or per serving. Values are compared with label claims and formulation targets. Consistency across batches confirms product stability, while deviations may indicate oxidation or storage-related degradation.

This assay quantifies beta-carotene, a provitamin A carotenoid that converts to active vitamin A in the body. It measures beta-carotene content in food and supplement samples using HPLC to support accurate labeling and formulation consistency.

Samples are extracted under light-protected conditions to preserve beta-carotene integrity. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity beta-carotene standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mcg per 100 g or per serving, and may be converted to IU based on equivalency factors. The values are compared with label claims and formulation goals. Consistent levels confirm stability, while variation may reflect degradation due to light or heat exposure.

This assay quantifies retinyl acetate, a stable and commonly used supplemental form of vitamin A, in food and supplement samples. It measures active vitamin A content using HPLC to ensure proper dosing and verify label claims in fortified products.

Samples are extracted under light-protected and oxygen-controlled conditions to preserve retinyl acetate. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity retinyl acetate standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in IU or mcg per 100 g or per serving. The values are compared with formulation targets and label claims. Consistent results confirm product stability, while variation may indicate degradation due to oxidation or improper storage.

This test quantifies retinyl palmitate, the most common esterified storage form of Vitamin A used in dietary supplements and fortified foods, using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Retinyl palmitate is a fat-soluble retinoid that is hydrolyzed in the body to retinol and subsequently converted to the active forms required for vision, immune response, and cellular differentiation. Results are reported in IU or mcg RAE (Retinol Activity Equivalents) per serving or per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and subjected to saponification using an ethanolic potassium hydroxide solution under heat to hydrolyze retinyl palmitate to free retinol, followed by liquid-liquid extraction with an organic solvent such as hexane. Alternatively, direct extraction without saponification may be used to quantify the intact ester form. The extract is evaporated, reconstituted in mobile phase, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 325 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified retinyl palmitate or retinol reference standard. All sample preparation steps are conducted under amber or reduced-light conditions to prevent photodegradation of the retinoid.

Retinyl palmitate is the predominant form of preformed Vitamin A in supplement formulations and must be accurately quantified to substantiate label claims expressed in IU or mcg RAE and to ensure compliance with FDA cGMP requirements. HPLC-UV at 325 nm provides the selectivity needed to resolve retinyl palmitate and retinol from carotenoids and other fat-soluble vitamins that may be present in multi-ingredient formulations, delivering reliable potency data across softgels, tablets, and powders.

This assay quantifies vitamin A as retinol, the active form of vitamin A, in food and supplement samples. It measures retinol content using HPLC to verify potency and ensure accurate dosing in formulations targeting skin, immune, and eye health.

Samples are extracted under light-protected and oxygen-controlled conditions to prevent degradation. The extract is analyzed by HPLC with detection based on specific mass transitions. Calibration with high-purity retinol standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in IU or mcg per 100 g or per serving. The values are compared with label claims and regulatory targets. Consistent results confirm formulation stability, while variability may suggest oxidation or sensitivity to storage conditions.

This assay quantifies adenosylcobalamin, one of the biologically active forms of vitamin B12, in food and supplement samples. It measures adenosylcobalamin content using LC-MS/MS to ensure potency in formulations targeting cellular energy and neurological health.

Samples are extracted under light- and temperature-controlled conditions to preserve adenosylcobalamin. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity adenosylcobalamin standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mcg per 100 g or per serving. The values are compared with label claims and formulation targets. Consistent results confirm product stability, while variation may indicate degradation or improper storage conditions.

This assay quantifies cyanocobalamin, the synthetic and most stable form of vitamin B12, in food and supplement samples. It measures cyanocobalamin content using LC-MS/MS to verify label claims and ensure accurate dosing in fortified products.

Samples are extracted under controlled conditions to preserve cyanocobalamin integrity. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity cyanocobalamin standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mcg per 100 g or per serving. The values are assessed against label claims and formulation targets. Consistency across batches confirms stability and dosing accuracy, while deviations may suggest degradation or quality issues.

This assay quantifies hydroxycobalamin, a natural and bioavailable form of vitamin B12, in food and supplement samples. It measures hydroxycobalamin content using LC-MS/MS to ensure accurate dosing in formulations focused on metabolic and nervous system support.

Samples are extracted under light-protected and temperature-controlled conditions to preserve hydroxycobalamin. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity hydroxycobalamin standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mcg per 100 g or per serving. The values are compared with label claims and formulation specifications. Stable values across batches confirm product quality, while deviations may indicate sensitivity to processing or storage conditions.

This test quantifies methylcobalamin, the active coenzyme form of Vitamin B12, in dietary supplements and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Methylcobalamin is preferred in premium supplement formulations for its direct bioavailability and role in neurological function and methionine synthesis. LC-MS/MS is required for this analysis due to its ability to specifically identify and quantify methylcobalamin at the trace levels present in supplement matrices, distinguishing it from other cobalamin forms such as cyanocobalamin and adenosylcobalamin. Results are reported in mcg per serving or mcg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using an aqueous buffer or methanol-water solvent system under low-light conditions to minimize photodegradation of the cobalamin compound. The clarified extract 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. Specific precursor-to-product ion transitions unique to methylcobalamin are monitored for quantification and confirmation. Quantification is performed against a multi-point calibration curve prepared from a certified methylcobalamin reference standard, with a stable isotope-labeled internal standard used to correct for matrix effects and recovery variability.

Methylcobalamin is present at microgram-per-serving levels in supplements, requiring a highly sensitive and selective method to achieve accurate quantification. Unlike HPLC-UV, LC-MS/MS can unambiguously distinguish methylcobalamin from structurally similar cobalamin forms (cyanocobalamin, hydroxocobalamin, adenosylcobalamin) that may co-elute under chromatographic conditions, making it the method of choice for label claim verification in products where the specific form of B12 is a key differentiator.

This assay quantifies total vitamin B12, including various forms like Adenosylcobalamin, Hydroxycobalamin, Methylcobalamin, and Cyanocobalamin, in food and supplement samples. It measures total B12 content using LC-MS/MS to verify nutritional claims and ensure consistent dosing.

Samples are extracted and treated to release all cobalamin forms. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with certified cobalamin standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mcg per 100 g or per serving. Values are compared with formulation targets and label claims. Consistent levels across batches confirm manufacturing accuracy, while deviations may signal formulation or raw material issues.

This assay quantifies amygdalin (commonly referred to as vitamin B17 or laetrile) in food and supplement samples. It measures amygdalin content using LC-MS/MS or HPLC to support consistency in niche formulations where this compound is included.

Samples are extracted under controlled conditions to preserve amygdalin. The extract is analyzed by LC-MS/MS or HPLC with detection based on compound-specific absorbance or mass transitions. Calibration with high-purity amygdalin standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. Values are assessed against formulation targets. Due to its controversial regulatory status, consistent and transparent reporting is critical for brands using amygdalin in functional or alternative wellness products.

This assay quantifies vitamin B1 (benfotiamine) in food and supplement samples. It involves converting benfotiamine to its biologically active form and measuring total thiamine content using LC-MS/MS. Accurate measurement is essential for verifying nutritional claims and ensuring formulation consistency.

Samples are extracted under conditions that convert benfotiamine into thiamine for analysis. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity thiamine standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. The values are compared with expected targets and label claims, and consistent results across batches confirm stable product formulation. Significant deviations may indicate processing or storage issues.

This test quantifies Vitamin B1 in the form of thiamine hydrochloride (thiamine HCl) — the most common supplemental form of this essential water-soluble B vitamin — in dietary supplements and raw materials using High-Performance Liquid Chromatography (HPLC). Thiamine is a critical coenzyme in carbohydrate metabolism and plays an essential role in nerve conduction and energy production. Accurate potency verification is necessary for label claim substantiation and for confirming that the declared amount of this essential vitamin 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 dissolved in a dilute acidic aqueous solvent to ensure complete dissolution of the thiamine HCl. 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 254 nm or fluorescence detection following post-column oxidation to the fluorescent thiochrome derivative, depending on the validated method. Quantification is performed against a multi-point external calibration curve prepared from a certified thiamine HCl reference standard. System suitability and quality control standards are run concurrently to confirm method accuracy and precision throughout the analytical run.

Thiamine HCl is an essential vitamin with a defined daily value, and accurate HPLC quantification is necessary to confirm that dietary supplement products meet their declared label claims and comply with cGMP requirements under 21 CFR 111. HPLC provides the specificity needed to quantify thiamine HCl in multi-ingredient supplement matrices, distinguishing it from other B vitamins and excipients that may co-elute under less selective analytical conditions. Fluorescence detection via thiochrome derivatization offers enhanced sensitivity for low-dose formulations where UV detection may not provide sufficient signal-to-noise.

This test quantifies thiamine (Vitamin B1) in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Thiamine is an essential water-soluble vitamin that acts as a coenzyme in carbohydrate metabolism and is critical for proper nerve and muscle function. Results are reported in mg per serving or mg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a dilute acidic aqueous solvent with sonication or gentle heating to ensure complete solubilization of thiamine. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 254 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified thiamine reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Thiamine is a required nutrient declaration on supplement facts panels and must be accurately quantified to substantiate label claims and meet FDA cGMP requirements. HPLC-UV provides the specificity needed to resolve thiamine from co-formulants and degradation products in complex supplement matrices, offering a more reliable measure of potency than microbiological or colorimetric methods in multi-ingredient formulations.

This test quantifies riboflavin (Vitamin B2) in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Riboflavin is an essential water-soluble vitamin that serves as a precursor to the coenzymes FMN and FAD, which are central to cellular energy metabolism and antioxidant defense. Results are reported in mg per serving or mg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a dilute acidic aqueous solvent with sonication or heating to ensure complete solubilization of riboflavin. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 267 nm or by fluorescence detection as an alternative, and quantification is performed against a multi-point external calibration curve prepared from a certified riboflavin reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility.

Riboflavin is a required nutrient declaration on supplement facts panels and must be accurately quantified to substantiate label claims and meet FDA cGMP requirements. HPLC-UV provides the specificity needed to measure riboflavin in complex supplement matrices, distinguishing it from structurally related flavin compounds and co-formulants that may interfere with simpler colorimetric assays.

This assay quantifies riboflavin 5-phosphate, the active coenzyme form of vitamin B2, in food and supplement samples. It involves stabilizing and measuring the riboflavin 5-phosphate content using LC-MS/MS. Accurate measurement is essential for verifying bioavailable forms and formulation consistency.

Samples are extracted under light-protected conditions to preserve riboflavin 5-phosphate integrity. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. The values are compared with expected targets and label claims, and consistent results across batches confirm stable product formulation. Significant deviations may indicate processing or storage issues.

This test quantifies niacinamide (nicotinamide), the amide form of Vitamin B3, in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Niacinamide is widely used in supplements and functional foods for its role in energy metabolism and NAD+ biosynthesis, and accurate potency measurement is required for label claim verification and cGMP compliance. Results are reported in mg per serving or mg per gram, as applicable.

A representative sample is weighed and extracted using a dilute aqueous or acidic solvent with sonication or gentle heating to ensure complete solubilization of niacinamide. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 254 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified niacinamide reference standard. System suitability and QC samples are run concurrently to confirm method performance and accuracy.

Niacinamide is the predominant form of Vitamin B3 used in dietary supplements and is distinct from nicotinic acid (niacin) in both its biochemical behavior and its side effect profile — notably, it does not cause the flushing response associated with niacin. HPLC-UV provides the specificity required to quantify niacinamide independently of niacin and other co-formulants, ensuring label claim accuracy and supporting regulatory compliance across finished products and raw materials.

This test quantifies Vitamin B3, including both niacin (nicotinic acid) and niacinamide (nicotinamide) forms, in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Niacin is an essential water-soluble B vitamin critical for energy metabolism and cellular function, and accurate potency measurement is required for label claim verification and compliance with FDA and cGMP requirements. Results are reported in mg per serving or mg per gram, as applicable.

A representative sample is weighed and extracted using a dilute aqueous or acidic solvent (such as dilute hydrochloric acid or phosphate buffer) with sonication or heating to ensure complete solubilization of both niacin and niacinamide forms. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Separation and detection are performed using UV detection at approximately 254 nm. Quantification is performed against a multi-point external calibration curve prepared from certified reference standards for both niacin and niacinamide, with system suitability and QC samples run concurrently to confirm method performance.

Niacin and niacinamide are the two primary forms of Vitamin B3 used in dietary supplements and food fortification, and both must be accurately measured to confirm label claims. HPLC-UV is the standard analytical approach for B vitamin quantification, offering the specificity needed to distinguish between the two forms and the sensitivity required to detect low-dose formulations. Accurate potency data is essential for regulatory compliance, label claim substantiation, and ensuring consistent product quality across batches.

This test quantifies pantothenic acid (Vitamin B5) in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Pantothenic acid is an essential water-soluble vitamin required for the synthesis of coenzyme A and plays a central role in fatty acid metabolism and energy production. Results are reported in mg per serving or mg per gram, as applicable, to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a dilute aqueous buffer or acidic solvent with sonication to ensure complete solubilization of pantothenic acid. The clarified extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 200–210 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified pantothenic acid reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility.

Pantothenic acid is a required nutrient declaration on supplement facts panels and must be accurately quantified to substantiate label claims and meet FDA cGMP requirements. HPLC-UV provides the specificity and sensitivity needed to measure pantothenic acid in complex supplement matrices, distinguishing it from co-formulants and excipients that may interfere with simpler colorimetric or microbiological methods.

This test quantifies pyridoxine hydrochloride (Vitamin B6) in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Pyridoxine HCl is the most common supplemental form of Vitamin B6, an essential coenzyme involved in amino acid metabolism, neurotransmitter synthesis, and immune function. Results are reported in mg per serving or mg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a dilute acidic aqueous solvent with sonication to ensure complete solubilization of pyridoxine HCl. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 290 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified pyridoxine HCl reference standard. System suitability and QC samples are run concurrently to confirm method performance and inter-run reproducibility.

Vitamin B6 is a required nutrient declaration on supplement facts panels and must be accurately quantified to substantiate label claims and meet FDA cGMP requirements. HPLC-UV with UV detection at 290 nm provides the selectivity needed to resolve pyridoxine from other B vitamins and common excipients, ensuring accurate potency measurement across a range of supplement formats including tablets, capsules, and powders.

This assay quantifies pyridoxal, a bioactive form of vitamin B6, in food and supplement samples. It measures pyridoxal content using HPLC to verify accurate dosing in formulations designed for nervous system support and B6-complex blends.

Samples are extracted under stabilized, light-protected conditions. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity pyridoxal standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. The values are compared with formulation targets and label claims. Consistent results confirm bioactive stability and proper dosing, while variability may suggest degradation or instability in certain storage conditions.

This assay quantifies pyridoxal 5’-phosphate (PLP), the active coenzyme form of vitamin B6, in food and supplement samples. It involves stabilizing and measuring PLP content using LC-MS/MS to verify potency and ensure formulation consistency.

Samples are extracted under light- and temperature-controlled conditions to preserve PLP. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity PLP standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. Values are compared with label claims and expected formulation targets. Stable readings across batches confirm product consistency, while variability may suggest degradation or quality control issues.

This assay quantifies pyridoxamine, one of the biologically active forms of vitamin B6, in food and supplement samples. It measures pyridoxamine content using HPLC to verify formulation potency and support accurate labeling in B6-complex and metabolic health products.

Samples are extracted under controlled conditions to preserve pyridoxamine. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity pyridoxamine standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving. Values are compared with formulation targets and label claims. Consistent levels confirm ingredient integrity, while variability may indicate degradation or conversion during processing or storage.

This test quantifies biotin (Vitamin B7) in dietary supplements and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). Biotin is an essential water-soluble vitamin that acts as a coenzyme in carboxylation reactions involved in fatty acid synthesis, gluconeogenesis, and amino acid metabolism. LC-MS/MS is the preferred method for biotin quantification due to the low microgram-to-milligram dose levels used in supplements and the need to distinguish biotin from structurally similar compounds in complex matrices. Results are reported in mcg per serving or mcg per gram to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using an aqueous buffer or methanol-water solvent system with sonication to ensure complete solubilization of biotin. The clarified extract 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. Specific precursor-to-product ion transitions characteristic of biotin are monitored for quantification and confirmation. Quantification is performed against a multi-point calibration curve prepared from a certified biotin reference standard, with a stable isotope-labeled internal standard used to correct for matrix effects and ensure accurate recovery across sample types.

Biotin is commonly formulated across a wide dose range — from standard 30–300 mcg levels to high-dose 5,000–10,000 mcg products — requiring a method with both the sensitivity to detect low doses and the dynamic range to accurately quantify high-dose formulations. LC-MS/MS provides the selectivity to distinguish biotin from co-eluting matrix components and structurally related compounds that can interfere with HPLC-UV or microbiological methods, making it the most reliable approach for label claim verification across diverse supplement formats.