Phytochemical, Vitamin, and Actives testing

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 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 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 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 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 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 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 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.

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 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 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 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 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 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 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.