Phytochemical, Vitamin, and Actives testing

This assay quantifies folic acid, the synthetic supplemental form of vitamin B9, in food and supplement samples. It measures total folic acid content using LC-MS/MS to verify dosage and support nutritional claims in fortified products.

Samples are extracted and treated to stabilize folic acid. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity folic acid 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 to label claims and formulation targets. Consistent readings confirm accurate dosing, while deviations may indicate manufacturing or degradation issues.

This assay quantifies folinic acid (also known as 5-formyltetrahydrofolate), a metabolically active form of folate, in food and supplement samples. It measures folinic acid content using LC-MS/MS to confirm potency in advanced formulations for methylation and cellular health.

Samples are extracted under stabilized conditions to preserve folinic acid. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity folinic acid 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 to formulation targets and label claims. Stable results confirm consistent manufacturing, while variability may indicate instability during processing or storage.

This test quantifies L-methyltetrahydrofolic acid (5-MTHF), the active, bioavailable form of Vitamin B9 (folate), in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Unlike synthetic folic acid, 5-MTHF does not require enzymatic conversion and is the predominant circulating form of folate in the body, making accurate potency measurement critical for products targeting methylation support, prenatal health, and cardiovascular function. 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 a dilute phosphate buffer or acidic aqueous solvent under low-light conditions to minimize photodegradation of the folate compound. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 290–300 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified 5-MTHF reference standard. System suitability and QC samples are run concurrently, and all sample preparation steps are conducted under amber or reduced-light conditions to preserve analyte integrity.

5-MTHF is increasingly used in premium supplement formulations as a bioavailable alternative to synthetic folic acid, particularly for individuals with MTHFR gene variants who have reduced capacity to convert folic acid to its active form. Accurate quantification of 5-MTHF is essential to substantiate label claims, differentiate it from other folate forms in multi-ingredient products, and meet FDA cGMP requirements. HPLC-UV provides the specificity and sensitivity required to measure this light-sensitive compound reliably across capsule, tablet, and powder matrices.

This test quantifies ascorbic acid (Vitamin C) in dietary supplements, fortified foods, beverages, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Vitamin C is one of the most widely used dietary supplement ingredients and is essential for immune defense, collagen synthesis, and antioxidant activity. 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 solvent — typically metaphosphoric acid or oxalic acid solution — to stabilize ascorbic acid and prevent oxidative degradation during sample preparation. The extract is filtered, diluted to volume, and injected onto a reversed-phase C18 HPLC column. Detection is performed by UV at approximately 245–254 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified ascorbic acid reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and stability of the analyte throughout the analytical run.

Vitamin C is a required nutrient declaration on supplement facts panels and is among the most commonly label-claimed ingredients in the supplement industry. Accurate potency measurement is essential for regulatory compliance and consumer trust. HPLC-UV is preferred over titration methods for complex matrices because it provides the selectivity to distinguish ascorbic acid from interfering reducing agents and co-formulants, delivering reliable results across tablets, capsules, powders, and liquids.

This assay quantifies ascorbyl palmitate, a fat-soluble ester form of vitamin C, in food and supplement samples. It measures the stabilized vitamin C content using LC-MS/MS to verify potency and support accurate dosing in lipid-based or cosmetic formulations.

Samples are extracted using solvents optimized for fat-soluble compounds. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity ascorbyl palmitate 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 readings confirm formulation accuracy and ingredient stability, while deviations may suggest degradation or incorporation issues.

This test quantifies the vitamin C content of calcium ascorbate — a calcium salt of ascorbic acid commonly used as a buffered, stomach-friendly alternative to ascorbic acid in dietary supplements — by measuring its ascorbic acid equivalent content using High-Performance Liquid Chromatography (HPLC). Calcium ascorbate contains approximately 89–91% ascorbic acid by mass (theoretical), and potency is reported as ascorbic acid equivalents to align with standard vitamin C label claim conventions under 21 CFR 101. Accurate quantification ensures that the declared vitamin C content is present and supports compliance with nutritional labeling requirements and cGMP standards under 21 CFR 111. Results are reported in milligrams of ascorbic acid equivalents per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate aqueous diluent, typically a dilute acidic solution (e.g., metaphosphoric acid or oxalic acid) to stabilize ascorbic acid and prevent oxidative degradation during sample preparation. The solution is filtered and analyzed by reversed-phase HPLC on a C18 column with UV detection at 254 nm, the characteristic absorption maximum of ascorbic acid. Quantification is performed against a multi-point external calibration curve prepared from a certified L-ascorbic acid reference standard. Both L-ascorbic acid and, where applicable, dehydroascorbic acid (the oxidized form) may be monitored to assess total vitamin C activity. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

While calcium ascorbate can be characterized by titration or colorimetric methods, HPLC with UV detection provides superior specificity by chromatographically resolving ascorbic acid from potential degradation products, excipients, and matrix interferences. This is particularly important for finished supplement matrices where other antioxidants or reducing agents may interfere with non-specific assay methods. Reporting results as ascorbic acid equivalents ensures direct comparability with label claims and regulatory requirements for vitamin C content, supporting both raw material qualification and finished product release testing.

This assay quantifies tetrahexyldecyl ascorbate, a highly stable, lipid-soluble derivative of vitamin C, in cosmetic and supplement products. It measures active compound levels using LC-MS/MS to ensure targeted dosing in formulations designed for dermal absorption and oxidative stress support.

Samples are extracted using lipid-compatible solvents under light- and heat-controlled conditions. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity tetrahexyldecyl ascorbate 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 formulation goals. Consistent results confirm active ingredient stability and proper incorporation, while deviations may indicate formulation or raw material quality issues.

This test quantifies Vitamin D2 (ergocalciferol), an essential nutrient commonly added to dietary supplements and fortified foods, using LC-MS/MS for precise measurement. It is suitable for various matrices including raw materials, capsules, and powders. The method detects Vitamin D2 at concentrations as low as 1 ng/g, ensuring accurate assessment of product potency and compliance with labeling requirements.

Samples are first saponified with ethanolic potassium hydroxide to release ergocalciferol, followed by extraction with hexane. The extract is evaporated and reconstituted in methanol before analysis. LC-MS/MS is performed using electrospray ionization in positive mode, with multiple reaction monitoring (MRM) transitions specific to ergocalciferol. Quantification is achieved via an external calibration curve prepared from certified Vitamin D2 standards. Method accuracy and precision are verified through duplicate injections, spiked recovery tests, and inclusion of quality control samples in each batch.

Testing confirms label claims, ensures formulation accuracy, and supports regulatory compliance for Vitamin D–fortified products.

This assay quantifies vitamin D2 (ergocalciferol), the plant-derived form of vitamin D, in food and supplement samples. It measures active D2 content using LC-MS/MS to verify potency in fortified products and support accurate label claims.

Samples are extracted under light-protected and temperature-controlled conditions to preserve vitamin D2. The extract is analyzed by LC-MS/MS with detection based on specific mass transitions. Calibration with high-purity ergocalciferol 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 assessed against formulation targets and regulatory standards. Consistent results confirm ingredient stability, while variability may reflect light or heat degradation during processing or storage.

This test quantifies cholecalciferol (Vitamin D3) in dietary supplements, fortified foods, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Vitamin D3 is the preferred supplemental form of Vitamin D due to its superior bioavailability and potency relative to Vitamin D2, and it plays a critical role in calcium absorption, bone mineralization, and immune regulation. Results are reported in IU or mcg 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 release cholecalciferol from its matrix, followed by liquid-liquid extraction with an organic solvent such as hexane or diethyl ether. The extract is evaporated, reconstituted in mobile phase, and injected onto a normal-phase or reversed-phase HPLC column. Detection is performed by UV at approximately 265 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified cholecalciferol reference standard. All sample preparation steps are conducted under amber or reduced-light conditions to prevent photodegradation, and system suitability and QC samples are run concurrently to confirm method performance.

Vitamin D3 is one of the most widely consumed dietary supplement ingredients and is subject to strict label claim requirements under FDA cGMP regulations. Accurate potency measurement is essential given the narrow therapeutic window between adequate intake and potential toxicity at high doses. HPLC-UV provides the selectivity to distinguish cholecalciferol from Vitamin D2 and related sterols in complex supplement matrices, ensuring reliable quantification across softgels, tablets, powders, and oil-based formulations.

This assay quantifies alpha-tocopherol, the most active and bioavailable form of vitamin E, in food and supplement samples. It measures vitamin E content using HPLC to ensure accurate dosing and verify label claims in antioxidant-focused formulations.

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

Results are provided in IU or mg per 100 g or per serving. The values are compared with formulation targets and label claims. Stable values confirm proper formulation and storage, while significant loss may indicate degradation due to oxidative stress.

This assay quantifies alpha-tocopherol acetate, a stabilized ester form of vitamin E, in food, supplement, and cosmetic samples. It measures vitamin E acetate content using HPLC to verify potency and ensure accurate dosing in formulations requiring oxidative stability.

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

Results are provided in IU or mg per 100 g or per serving. The values are compared with formulation targets and label claims. Stable values confirm ingredient integrity and resistance to oxidation, while loss may suggest hydrolysis or improper storage.

This assay quantifies alpha-tocopheryl succinate, a stable esterified form of vitamin E, in food and supplement samples. It measures vitamin E content using HPLC to ensure accurate dosing in formulations focused on oxidative stress and cellular defense.

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

Results are provided in IU or mg per 100 g or per serving. The values are compared with formulation targets and label claims. Consistent levels confirm ingredient stability and proper delivery, while loss may suggest hydrolysis or degradation over time.

This assay quantifies mixed tocopherols—typically including alpha-, beta-, gamma-, and delta-tocopherol—in food and supplement samples. It measures the individual tocopherol components using HPLC to ensure balanced formulation and support full-spectrum vitamin E labeling.

Samples are extracted under light- and oxygen-controlled conditions to protect all tocopherol forms. The extract is analyzed by HPLC with separation and detection of each tocopherol based on its unique absorbance. Calibration with individual high-purity tocopherol standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving for each tocopherol component. Values are assessed individually and in total against formulation targets and label claims. This panel helps confirm tocopherol balance and stability across batches, while any imbalances may indicate degradation or formulation drift.

This assay quantifies beta-tocopherol, one of the naturally occurring forms of vitamin E, in food and supplement samples. It measures beta-tocopherol content using HPLC to support accurate labeling in full-spectrum or mixed tocopherol formulations.

Samples are extracted under light- and oxygen-controlled conditions to prevent oxidative degradation. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity beta-tocopherol 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 label claims and formulation targets. Consistent levels confirm proper formulation and antioxidant preservation, while variation may indicate oxidation or ingredient imbalance.

This assay quantifies delta-tocopherol, a lesser-known but potent form of vitamin E, in food and supplement samples. It measures delta-tocopherol content using HPLC to support precise formulation in mixed tocopherol or antioxidant-focused products.

Samples are extracted under oxygen- and light-protected conditions to preserve tocopherol stability. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity delta-tocopherol 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 assessed against formulation goals and label claims. Stable results confirm ingredient preservation and dosing accuracy, while variation may reflect oxidative degradation or formulation inconsistency.

This assay quantifies delta-tocotrienol, one of the potent and fast-acting members of the tocotrienol family within the vitamin E complex. It measures delta-tocotrienol content in food and supplement samples using HPLC to support precise formulation in advanced antioxidant products.

Samples are extracted under light- and oxygen-controlled conditions to preserve the integrity of tocotrienols. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity delta-tocotrienol 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 assessed against formulation targets and label claims. Consistent levels confirm stability and proper delivery, while loss may suggest oxidative degradation or improper storage.

This assay quantifies gamma-tocopherol, a major dietary form of vitamin E with unique antioxidant properties, in food and supplement samples. It measures gamma-tocopherol content using HPLC to ensure precise formulation and support accurate label claims in mixed tocopherol products.

Samples are extracted under controlled light and oxygen conditions to prevent oxidation. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity gamma-tocopherol 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 levels confirm product quality and antioxidant protection, while variability may signal oxidation or ingredient degradation.

This assay quantifies gamma-tocotrienol, a highly bioactive member of the vitamin E tocotrienol family, in food and supplement samples. It measures gamma-tocotrienol content using HPLC to ensure potency and formulation accuracy in advanced antioxidant products.

Samples are extracted under light- and oxygen-controlled conditions to preserve tocotrienol stability. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity gamma-tocotrienol 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 to formulation goals and label claims. Stable results confirm ingredient preservation and formulation accuracy, while variation may indicate degradation or sourcing inconsistencies.

This assay quantifies mixed tocotrienols—typically including alpha-, beta-, gamma-, and delta-tocotrienol—in food and supplement samples. It measures each form individually using HPLC to verify label claims and ensure proper formulation in advanced antioxidant products.

Samples are extracted under light- and oxygen-protected conditions to preserve all tocotrienol isomers. The extract is analyzed by HPLC with separation and detection of each tocotrienol form based on unique absorbance. Calibration with individual high-purity tocotrienol standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving for each tocotrienol component and as a total tocotrienol value. Values are assessed against formulation targets and label claims. Consistent results confirm isomer balance and ingredient stability, while variation may indicate degradation or sourcing issues.

This assay quantifies natural vitamin E (D-alpha-tocopherol), derived from plant-based oils, in food and supplement samples. It measures the bioactive natural form using HPLC to ensure accurate dosing and support label claims for premium, naturally sourced products.

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

Results are provided in IU or mg per 100 g or per serving. The values are compared with formulation targets and label claims. Consistent results confirm use of high-quality, natural-sourced vitamin E, while variability may indicate degradation or improper storage.

This test quantifies the full panel of Vitamin E homologs — including alpha-, beta-, gamma-, and delta-tocopherols and, where present, alpha-, beta-, gamma-, and delta-tocotrienols — in dietary supplements, oils, botanical extracts, and raw materials using High-Performance Liquid Chromatography (HPLC). Vitamin E is a family of eight structurally related fat-soluble antioxidants, and the relative composition of individual homologs varies significantly between natural sources (e.g., mixed tocopherols from soy or sunflower) and synthetic forms (dl-alpha-tocopherol). Individual homolog quantification provides a more complete and accurate measure of Vitamin E potency and source authenticity than total tocopherol assays alone. Results are reported in milligrams per gram or per serving for each individual homolog, with total Vitamin E activity expressed in IU or mg alpha-tocopherol equivalents as applicable.

A representative sample is accurately weighed and extracted using an organic solvent such as hexane or ethanol, with saponification applied where necessary to release esterified tocopherols from complex food or supplement matrices. The extract is filtered and analyzed by normal-phase or reversed-phase HPLC on a silica or C18 column, with fluorescence detection (excitation ~295 nm, emission ~330 nm) for high sensitivity, or UV detection at approximately 292 nm as an alternative. Individual tocopherol and tocotrienol homologs are resolved chromatographically and quantified against a multi-point external calibration curve prepared from certified reference standards for each target homolog. System suitability and quality control standards are run concurrently to confirm method accuracy and precision throughout the analytical run.

The biological activity and antioxidant potency of Vitamin E varies significantly between homologs — alpha-tocopherol has the highest biological activity, while gamma-tocopherol and tocotrienols offer distinct and complementary antioxidant and anti-inflammatory properties. Full panel quantification by HPLC is therefore essential for accurately characterizing the Vitamin E composition of natural mixed tocopherol ingredients, confirming that the declared form and potency are present, and distinguishing natural-source Vitamin E (d-alpha-tocopherol) from synthetic forms (dl-alpha-tocopherol). This level of analytical detail supports label claim accuracy, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies synthetic vitamin E (DL-alpha-tocopherol), a racemic mixture of eight stereoisomers, in food and supplement samples. It measures total synthetic vitamin E content using HPLC to ensure proper dosing and verify label claims in cost-effective formulations.

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

Results are provided in IU or mg per 100 g or per serving. The values are compared with formulation targets and regulatory limits. Consistent results confirm manufacturing accuracy, while deviations may suggest oxidation or improper handling of the synthetic form.

This assay quantifies total tocotrienols—typically including alpha-, beta-, gamma-, and delta-tocotrienol—in food and supplement samples. It measures each tocotrienol form using HPLC to ensure full-spectrum coverage and verify label claims in advanced antioxidant formulations.

Samples are extracted under light- and oxygen-protected conditions to prevent tocotrienol degradation. The extract is analyzed by HPLC with separation and detection of each isomer at a specific wavelength. Calibration with individual high-purity tocotrienol standards, along with internal standard correction and duplicate runs, ensures reliable quantification.

Results are provided in mg per 100 g or per serving for each tocotrienol isomer and total tocotrienol content. The values are assessed against formulation targets and label claims. This panel helps verify ingredient balance, support full-spectrum antioxidant claims, and detect any degradation across storage or production.

This assay quantifies vitamin K1—also known as phylloquinone or phytonadione—in food and supplement samples. It measures vitamin K1 content using HPLC to ensure accurate dosing and verify label claims in formulations supporting cardiovascular and skeletal health.

Samples are extracted under light- and oxygen-protected conditions to preserve the sensitive K1 molecule. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity phylloquinone 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 results confirm ingredient stability, while variation may reflect degradation due to light, pH, or improper handling.

This assay quantifies menaquinone-4 (MK-4), a short-chain form of vitamin K2, in food and supplement samples. It measures MK-4 content using HPLC to verify label claims and ensure potency in formulations designed to support skeletal and cardiovascular health.

Samples are extracted under light- and oxygen-protected conditions to prevent MK-4 degradation. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity MK-4 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 formulation targets and label claims. Consistent results confirm ingredient stability and bioactive dosing, while variability may indicate oxidation or instability during storage or processing.

This assay quantifies menaquinone-7 (MK-7), a long-chain and highly bioavailable form of vitamin K2, in food and supplement samples. It measures MK-7 content using HPLC to verify potency and ensure extended-action support in bone and vascular formulations.

Samples are extracted under light- and oxygen-controlled conditions to protect MK-7 from degradation. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity MK-7 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 formulation targets and label claims. Stable results confirm ingredient preservation and sustained-release potential, while degradation may suggest sensitivity to light, pH, or improper storage.

This assay quantifies menadione (vitamin K3), a synthetic precursor to vitamins K1 and K2, in food, feed, or supplement samples. It measures menadione content using HPLC to verify formulation accuracy in non-human products and certain fortified blends.

Samples are extracted under light- and oxygen-controlled conditions to preserve the compound’s integrity. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity menadione 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 specifications and safety thresholds. Accurate quantification is critical due to K3’s limited use in human nutrition and regulatory restrictions in many regions.

This assay quantifies synthetic vitamin K3 (menadione), a provitamin compound used in veterinary and non-human applications. It measures menadione content using HPLC to ensure precise formulation and compliance in feed-grade and specialty use products.

Samples are extracted under oxygen- and light-controlled conditions to preserve menadione’s structure. The extract is analyzed by HPLC with detection based on absorbance at a specific wavelength. Calibration with high-purity menadione 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 target specifications for feed or industrial use. Accurate measurement is essential due to regulatory restrictions on K3 in human supplements and the need for consistent dosing in non-human formulations.

This assay quantifies withanolide glycosides, the primary bioactive compounds in Ashwagandha (Withania somnifera). Using HPLC, it provides a quantitative profile of withanolides to verify standardization in root or extract-based formulations and support label claims for stress, hormone, and energy balance.

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

Results are reported in mg per g or per serving, typically as total withanolides. Values are assessed against formulation targets and declared standardization levels (e.g., 5% or 10% withanolides) to confirm potency and detect adulteration or low-quality sourcing.

This test quantifies xylo-oligosaccharides (XOS) in raw materials and finished dietary supplement or functional-food products using High-Performance Liquid Chromatography (HPLC). XOS are short-chain carbohydrates composed principally of β-(1→4)-linked xylose units, commonly including xylobiose (X2), xylotriose (X3), xylotetraose (X4), xylopentaose (X5), and higher-degree-of-polymerization xylo-oligosaccharides. They are typically manufactured by controlled enzymatic or hydrolytic conversion of xylan-rich plant materials, such as corn cobs, sugarcane bagasse, wheat bran, or hardwood hemicellulose. The assay separates and quantifies individual XOS fractions, and may report total XOS as the sum of the validated oligomer fractions. Free xylose and other low-molecular-weight sugars can also be measured separately when required, helping distinguish the intended oligosaccharide content from residual monosaccharide. Results are reported as % w/w, g/100 g, mg/g, or mg per serving.

A representative sample is accurately weighed and dissolved or extracted in purified water under conditions validated to achieve complete recovery of soluble XOS without hydrolysis of the oligomer distribution. The sample is clarified by centrifugation and filtration, typically through a 0.20 μm or 0.45 μm membrane, before analysis. The prepared extract is analyzed by HPLC using a carbohydrate-appropriate separation column, such as an amino-bonded, ligand-exchange, size-exclusion, or dedicated oligosaccharide column, selected to resolve xylose from xylobiose, xylotriose, xylotetraose, xylopentaose, and other validated XOS fractions. Detection is generally performed using refractive-index (RI) detection, although evaporative light-scattering detection (ELSD), charged-aerosol detection (CAD), or another validated universal carbohydrate detector may be used where greater selectivity or sensitivity is needed. Quantification is performed using multi-point calibration curves prepared from qualified, purity-corrected reference standards for xylose and the individual XOS oligomers, where commercially available. Total XOS is calculated as the sum of the validated oligomer peaks, excluding free xylose unless the product specification explicitly includes it. Method blanks, system-suitability standards, duplicate preparations, and matrix-spike or recovery quality-control samples are evaluated with each run to verify resolution, accuracy, precision, and calibration performance.

XOS ingredients are inherently heterogeneous mixtures rather than single chemical entities; therefore, total carbohydrate testing alone cannot verify the intended oligosaccharide composition or distinguish XOS from free xylose, glucose, sucrose, maltodextrins, or other added carbohydrates. HPLC provides direct chromatographic separation of the individual oligomer fractions and enables quantification of the XOS profile most relevant to ingredient quality and product specification. Monitoring both the individual degrees of polymerization and residual monosaccharides helps confirm manufacturing consistency, detect degradation or adulteration, and support accurate declaration of XOS content. The method supports raw-material qualification, formulation control, label-claim substantiation where applicable, and dietary supplement cGMP quality-control requirements under 21 CFR 111.

This assay quantifies yohimbine, an alkaloid derived from the bark of Pausinystalia yohimbe, commonly used in fat-burning and stimulant-based supplements. Using HPLC, it verifies yohimbine content in capsules and herbal extracts to ensure proper dosing, label compliance, and regulatory safety.

Samples are extracted using alcohol-based or acidic aqueous solvents and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified yohimbine 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 and detect over-formulation, underdosing, or substitution with non-standardized material.

This assay quantifies zeaxanthin, a key dietary carotenoid concentrated in the retina and often combined with lutein in vision-support supplements. Using HPLC, it measures zeaxanthin content in raw materials and finished products to confirm label claims, ensure potency, and maintain ingredient quality.

Samples are extracted using organic solvents (typically ethanol or hexane-based), then analyzed by HPLC with UV-Vis detection at a carotenoid-specific wavelength (typically ~450 nm). Quantification is performed using certified zeaxanthin standards, with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in mg per g or per serving. Values are assessed against formulation targets and label claims to confirm dosing, detect degradation, and verify ingredient authenticity.

This test confirms the botanical identity of Sceletium tortuosum (Kanna), including the proprietary Zembrin extract, in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic mesembrine alkaloid fingerprint of the sample is compared against a certified Sceletium tortuosum reference standard to confirm species authenticity and detect substitution with other botanical materials or synthetic alkaloid preparations. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or acidified methanol and applied alongside a certified Sceletium tortuosum reference standard onto an HPTLC silica gel plate. The plate is developed in a validated solvent system, examined under UV at 254 nm and 366 nm, and derivatized with Dragendorff or anisaldehyde-sulfuric acid reagent to visualize the characteristic mesembrine alkaloid bands. The resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Sceletium tortuosum is a niche, high-value botanical with a limited number of authenticated suppliers, making identity verification critical for brands sourcing this ingredient. The mesembrine alkaloid profile is species-specific and serves as the key differentiator for HPTLC-based identity confirmation, supporting supplier qualification and cGMP compliance under 21 CFR 111.