Phytochemical, Vitamin, and Actives testing

This test quantifies lecithin content in raw materials, phospholipid concentrates, and dietary supplements by measuring total phosphorus using a validated colorimetric assay. Lecithin is a complex mixture of phospholipids — primarily phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, and phosphatidic acid — derived from botanical sources such as soy (Glycine max) or sunflower (Helianthus annuus). Because each phospholipid molecule contains one phosphorus atom, total phosphorus content provides a direct and reproducible measure of total phospholipid (lecithin) concentration. This approach is widely used for lecithin standardization and quality control in the food, pharmaceutical, and dietary supplement industries. Results are reported as a percentage of total phospholipids or in milligrams of phospholipid per gram, calculated from total phosphorus content using the appropriate stoichiometric conversion factor.

A representative sample is accurately weighed and subjected to complete acid digestion — typically using a perchloric acid or sulfuric acid/hydrogen peroxide wet ashing procedure — to convert all organic phosphorus to inorganic orthophosphate. The digested solution is reacted with a colorimetric reagent system, most commonly the ammonium molybdate/ascorbic acid (molybdenum blue) method or the Fiske-SubbaRow method, to form a colored phosphomolybdate complex with absorbance measured spectrophotometrically at 820 nm (molybdenum blue) or 660 nm (Fiske-SubbaRow). Total phosphorus concentration is calculated from a multi-point calibration curve prepared from a certified inorganic phosphate reference standard (e.g., potassium dihydrogen phosphate). Lecithin content is calculated from total phosphorus using the mean molecular weight of the phospholipid mixture and the stoichiometric ratio of one phosphorus atom per phospholipid molecule. All measurements are performed in triplicate and averaged for final quantification.

The total phosphorus colorimetric assay is a well-established, cost-effective, and reproducible method for lecithin quantification that exploits the stoichiometric relationship between phosphorus content and phospholipid concentration. It provides a practical alternative to HPLC-based phospholipid class profiling for routine quality control and raw material release testing where total phospholipid content — rather than individual phospholipid class distribution — is the primary specification parameter. The method is applicable to both soy and sunflower lecithin sources and is aligned with established industry and pharmacopeial approaches for phospholipid quantification, supporting label claim substantiation and cGMP compliance under 21 CFR 111.

This test confirms the botanical identity of lemon balm leaf extract raw materials using High-Performance Thin-Layer Chromatography (HPTLC). Lemon balm is derived from Melissa officinalis L. (Lamiaceae), a widely used medicinal herb recognized in major pharmacopoeias including the European Pharmacopoeia (Ph. Eur.) and the German Commission E monographs. Lemon balm leaf extract is used in dietary supplements for its calming, anxiolytic, sleep-supportive, and cognitive function properties, with rosmarinic acid, caffeic acid, luteolin glycosides (including luteolin-7-O-glucoside), and hydroxycinnamic acid derivatives serving as the principal marker compounds. Rosmarinic acid is the most abundant and pharmacologically significant phenolic acid in lemon balm and serves as the primary quantitative marker for extract standardization. HPTLC identity testing generates a characteristic chromatographic fingerprint of the lemon balm leaf extract — encompassing its hydroxycinnamic acid and flavonoid profiles — that is compared against an authenticated reference standard to confirm species identity and detect potential substitution or adulteration with morphologically similar Lamiaceae species such as spearmint (Mentha spicata), peppermint (Mentha × piperita), or other mint-family herbs.

A representative sample of lemon balm leaf extract raw material is accurately weighed and dissolved or extracted in an appropriate solvent system (e.g., methanol or 70% ethanol) to produce a solution enriched in the characteristic hydroxycinnamic acid and flavonoid constituents. The extract is applied alongside a certified Melissa officinalis reference standard and, where applicable, reference standards for rosmarinic acid and caffeic acid, onto a silica gel 60 F₂₅₄ HPTLC plate using an automated sample applicator to ensure precise, reproducible band placement. The plate is developed in a validated solvent system (e.g., ethyl acetate/formic acid/glacial acetic acid/water) optimized for resolution of lemon balm's characteristic phenolic acid and flavonoid markers. After development, the plate is dried and visualized under UV light at 254 nm and 366 nm, followed by derivatization with an appropriate spray reagent (e.g., Natural Products Reagent A / PEG 400 or anisaldehyde-sulfuric acid) and heating to reveal the full characteristic fingerprint. Natural Products Reagent A is particularly effective for lemon balm, producing characteristic blue-green fluorescence for rosmarinic acid and related hydroxycinnamic acids under 366 nm UV illumination. The resulting chromatographic pattern — including the positions, colors, and relative intensities of all characteristic zones — is compared with the authenticated reference standard. Identity is confirmed when the sample fingerprint is concordant with the reference. The method is performed in accordance with USP General Chapter <203> (HPTLC) and/or AHPA/AHP HPTLC reference standards.

HPTLC is the preferred method for botanical identity testing of lemon balm leaf extract because it generates a holistic chromatographic fingerprint of the complex mixture of hydroxycinnamic acids (rosmarinic acid, caffeic acid) and flavonoids (luteolin glycosides) that together constitute the authentic chemical profile of Melissa officinalis. This multi-marker fingerprint approach is more discriminating than single-marker rosmarinic acid testing alone, as rosmarinic acid is present in numerous other Lamiaceae species, and the complete fingerprint pattern is required to distinguish authentic lemon balm from potential adulterants or substitutes. The characteristic fluorescence behavior of lemon balm's phenolic constituents under Natural Products Reagent A derivatization provides a highly visual and distinctive identity confirmation. Botanical identity verification is a mandatory requirement under 21 CFR 111.75 for dietary supplement raw material qualification, and HPTLC identity testing is explicitly recognized as an appropriate identity testing methodology in USP General Chapter <203>.

This test quantifies L-ergothioneine, a naturally occurring thiohistidine betaine amino acid found at high concentrations in mushrooms and certain other fungi, in dietary supplements and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). L-ergothioneine has attracted significant research interest for its potent antioxidant properties, cellular protective effects, and its designation by some researchers as a potential longevity vitamin. 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 methanol-water solvent system with sonication to ensure complete solubilization of L-ergothioneine 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 254 nm, utilizing the characteristic UV absorbance of the thiohistidine chromophore. Quantification is performed against a multi-point external calibration curve prepared from a certified L-ergothioneine reference standard, with system suitability and QC samples run concurrently to confirm method accuracy and reproducibility.

L-ergothioneine is an emerging premium ingredient in the supplement market, and accurate quantification is essential for verifying the potency of mushroom-derived extracts and standardized ergothioneine ingredients. HPLC-UV at 254 nm provides adequate selectivity for ergothioneine in most supplement matrices; however, for complex or low-concentration samples, LC-MS/MS may be considered as a confirmatory approach to resolve any co-eluting matrix interferences and ensure label claim accuracy.

This assay quantifies L-glutamine, a conditionally essential amino acid critical for recovery and intestinal barrier support. Using LC-MS/MS, it measures glutamine content in raw materials and finished products to verify potency, ensure label accuracy, and confirm consistency across batches.

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

Results are reported in mg per g or per serving. Values are compared to label claims and formulation targets to confirm potency, detect degradation, or identify underformulation.

This test measures the concentration of L-glutathione, a key antioxidant, in raw materials, capsules, powders, and finished products to ensure potency, purity, and label compliance. Using high-performance liquid chromatography (HPLC), the method accurately quantifies L-glutathione at low detection limits, supporting quality control in antioxidant and cellular support formulations.

Samples are prepared by extraction with an appropriate solvent to release L-glutathione from the matrix. The extract is then analyzed using HPLC equipped with UV or diode-array detection to separate L-glutathione from other thiol-containing compounds. Quantification is achieved by comparing sample peak areas to a calibration curve constructed from certified L-glutathione reference standards. Method accuracy and precision are confirmed through replicate injections, quality control samples, and spike recovery assessments.

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

This test quantifies L-Glycine, an important amino acid, in raw materials, powders, capsules, and finished products using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method offers precise and sensitive measurement down to low microgram per gram levels, ensuring product quality and compliance with formulation specifications.

Samples are extracted with an aqueous solvent and filtered prior to analysis. L-Glycine is separated by liquid chromatography and detected using tandem mass spectrometry in multiple reaction monitoring (MRM) mode targeting specific glycine transitions. Quantification is performed using a calibration curve constructed from certified reference standards. Method accuracy and precision are verified through duplicate injections and recovery assessment of spiked samples.

Results are reported in mg/g (raw material) or mg/serving (finished product). Testing confirms label accuracy, ensures consistent potency, and verifies raw material purity.

This test quantifies L-histidine — a semi-essential amino acid that serves as a precursor to histamine and carnosine, and plays important roles in immune regulation, oxygen transport via hemoglobin, and tissue repair — in dietary supplements, protein blends, amino acid formulations, and raw materials using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Accurate quantification is important for label claim verification in amino acid and protein supplement products. LC-MS/MS provides the sensitivity and compound-specific selectivity needed to accurately quantify L-histidine in complex amino acid matrices, where its unique imidazole side chain can present challenges for certain chromatographic methods. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved or hydrolyzed in a suitable aqueous solvent. An isotopically labeled internal standard (e.g., ¹³C- or ²H-labeled L-histidine) is added prior to sample preparation to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase or HILIC LC-MS/MS, with detection by electrospray ionization (ESI) in positive ion mode using multiple reaction monitoring (MRM) transitions specific to L-histidine. Quantification is performed against a multi-point external calibration curve prepared from a certified L-histidine reference standard. Quality control samples at multiple concentration levels are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

L-histidine's imidazole side chain confers a unique basic character and chromatographic behavior that can make it challenging to retain and resolve on standard reversed-phase columns under typical amino acid analysis conditions. LC-MS/MS with MRM detection provides the compound-specific selectivity and sensitivity needed to unambiguously quantify L-histidine in complex protein and amino acid matrices regardless of chromatographic challenges, ensuring accurate label claim verification. This level of analytical rigor supports raw material qualification and cGMP compliance under 21 CFR 111.

This test confirms the botanical identity of licorice root raw materials using High-Performance Thin-Layer Chromatography (HPTLC). Licorice root is derived primarily from Glycyrrhiza glabra L. (Spanish licorice), Glycyrrhiza uralensis Fisch. (Chinese licorice), or Glycyrrhiza inflata Bat., all of which are recognized as official sources in major pharmacopoeias including the United States Pharmacopeia (USP), European Pharmacopoeia (Ph. Eur.), and the Chinese Pharmacopoeia. Licorice root is widely used in dietary supplements for its adaptogenic, anti-inflammatory, and demulcent properties, with glycyrrhizin (glycyrrhizinic acid) and its aglycone glycyrrhetinic acid being the principal bioactive and marker compounds. HPTLC identity testing generates a characteristic chromatographic fingerprint of the licorice root extract — including glycyrrhizin, glycyrrhetinic acid, liquiritin, and other flavonoid and triterpenoid markers — that is compared against an authenticated reference standard to confirm species identity and detect potential substitution or adulteration.

A representative sample of licorice root raw material is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or ethanol/water) to produce a total extract enriched in the characteristic triterpenoid saponins and flavonoid constituents. The extract is applied alongside a certified Glycyrrhiza spp. reference standard and, where applicable, reference standards for glycyrrhizin and glycyrrhetinic acid, onto a silica gel 60 F₂₅₄ HPTLC plate using an automated sample applicator to ensure precise, reproducible band placement. The plate is developed in a validated solvent system (e.g., ethyl acetate/formic acid/glacial acetic acid/water) optimized for resolution of licorice root's characteristic triterpenoid and flavonoid markers. After development, the plate is dried and visualized under UV light at 254 nm and 366 nm, followed by derivatization with an appropriate spray reagent (e.g., anisaldehyde-sulfuric acid or Natural Products Reagent A) and heating to reveal the full characteristic fingerprint. The resulting chromatographic pattern — including the positions, colors, and relative intensities of all characteristic zones — is compared with the authenticated reference standard. Identity is confirmed when the sample fingerprint is concordant with the reference. The method is performed in accordance with USP General Chapter <203> (HPTLC) and/or AHPA/AHP HPTLC reference standards.

HPTLC is the preferred method for botanical identity testing of licorice root because it generates a holistic chromatographic fingerprint of the complex mixture of triterpenoid saponins (glycyrrhizin, glycyrrhetinic acid) and flavonoids (liquiritin, isoliquiritin) that together constitute the authentic chemical profile of Glycyrrhiza spp. This multi-marker fingerprint approach is more discriminating than single-marker testing and is capable of detecting adulteration with morphologically similar roots (e.g., Glycyrrhiza echinata, Abrus precatorius) that would not be identified by visual inspection alone. Botanical identity verification is a mandatory requirement under 21 CFR 111.75 for dietary supplement raw material qualification, and HPTLC identity testing is explicitly recognized as an appropriate identity testing methodology in USP General Chapter <203> and FDA's Dietary Supplement Ingredient Advisory List guidance.

This test simultaneously identifies and quantifies four key bioactive secondary metabolites of lion's mane mushroom (Hericium erinaceus) — erinacine A (a cyathane diterpenoid from the mycelium) and hericenones A, C, and D (aromatic compounds from the fruiting body) — using a combined LC-MS/MS and HPLC approach. Erinacines (found exclusively in the mycelium) and hericenones (found in the fruiting body) are the two principal classes of bioactive compounds responsible for the nerve growth factor (NGF)-stimulating and neuroprotective activity attributed to H. erinaceus. Erinacine A is the most pharmacologically potent and well-characterized erinacine, with strong evidence for NGF induction, while hericenones A, C, and D are aromatic isoindolinone and related compounds that contribute to the fruiting body's neuroactive profile. Comprehensive panel quantification of both compound classes enables differentiation between mycelium-based and fruiting body-based ingredients, supports label claim verification, and provides a complete characterization of the neuroactive bioactive content of the ingredient. Results are reported in milligrams per gram or per serving for each individual compound.

A representative sample is accurately weighed and extracted using an appropriate organic solvent system (e.g., methanol or ethanol/water) optimized to capture both the diterpenoid erinacines and the aromatic hericenone fractions. For LC-MS/MS analysis of erinacine A, the extract is analyzed using reversed-phase chromatography with electrospray ionization (ESI) in positive ion mode, with multiple reaction monitoring (MRM) transitions selected for the characteristic precursor and product ions of erinacine A. For HPLC quantification of hericenones A, C, and D, the extract is analyzed by reversed-phase HPLC on a C18 column with UV detection at an appropriate wavelength (typically 210–280 nm) that provides optimal sensitivity for the hericenone chromophores. Quantification of erinacine A is performed against a multi-point external calibration curve prepared from a certified erinacine A reference standard. Hericenones A, C, and D are quantified against certified reference standards for each individual compound. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

The combined LC-MS/MS and HPLC approach is employed because erinacine A and the hericenones differ significantly in their chemical properties, concentrations, and optimal detection modes. Erinacine A — a complex diterpenoid present at relatively low concentrations — requires the compound-specific selectivity and sensitivity of LC-MS/MS with MRM detection for accurate quantification in complex mushroom matrices. Hericenones A, C, and D — aromatic compounds present at higher concentrations in fruiting body materials — are amenable to HPLC with UV detection, which provides a practical and cost-effective quantification approach. Critically, the simultaneous profiling of both erinacines (mycelium markers) and hericenones (fruiting body markers) enables definitive characterization of the ingredient's source composition, distinguishing pure fruiting body, pure mycelium, and blended materials — a distinction of major commercial and quality significance in the lion's mane supplement market. This supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This test confirms the identity of lion's mane mushroom (Hericium erinaceus) in raw materials, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Hericium erinaceus is a culinary and medicinal mushroom native to North America, Europe, and Asia, widely used in dietary supplements for its potential to support cognitive function, nerve growth factor (NGF) stimulation, and immune health. Its characteristic phytochemical profile includes hericenones (from the fruiting body) and erinacines (from the mycelium), along with polysaccharides and other secondary metabolites. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated H. erinaceus reference standard to confirm species identity and detect potential adulteration, substitution, or misidentification with related Hericium species or inferior fungal materials.

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

Lion's mane is subject to adulteration and species substitution, particularly given the existence of closely related Hericium species (e.g., H. coralloides, H. americanum) and the variability in quality between fruiting body and mycelium-based materials. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation that distinguishes authentic H. erinaceus from related species and non-fungal adulterants, offering a level of discriminatory power that single-marker potency assays cannot provide. 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 the enzymatic activity of lipase, which catalyzes the hydrolysis of triglycerides into free fatty acids and glycerol. Testing ensures potency, stability, and consistency of enzyme-containing products.

Samples are incubated with an appropriate triglyceride substrate under controlled pH and temperature conditions. The release of fatty acids is quantified using a colorimetric or titrimetric method. Enzyme activity is reported relative to standardized reference units.

Results are reported in Lipase Units (LU) per gram (raw materials) or per serving (finished products). Testing verifies label claims, confirms enzyme stability, and ensures batch-to-batch consistency.

This test quantifies L-isoleucine — one of the three branched-chain amino acids (BCAAs), alongside leucine and valine — in dietary supplements, protein blends, amino acid formulations, and raw materials using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). L-isoleucine plays a critical role in muscle protein synthesis, energy metabolism, and blood glucose regulation, and is a key component of BCAA and sports nutrition products. LC-MS/MS provides the sensitivity and specificity needed to accurately quantify L-isoleucine and distinguish it from its structural isomers, including L-leucine and L-norvaline, in complex amino acid matrices. Results are reported as a percentage or in milligrams per serving to support label claim verification and cGMP compliance.

A representative sample is accurately weighed and dissolved or hydrolyzed in a suitable aqueous solvent. An isotopically labeled internal standard (e.g., ¹³C- or ²H-labeled L-isoleucine) is added prior to sample preparation to correct for matrix effects and recovery variability. The sample is filtered and injected onto a reversed-phase or HILIC LC column for chromatographic separation. Detection is performed by electrospray ionization (ESI) in positive ion mode, with multiple reaction monitoring (MRM) transitions selected to provide highly specific quantification of L-isoleucine. Quantification is performed against a multi-point external calibration curve prepared from a certified L-isoleucine reference standard. Quality control samples at multiple concentration levels are analyzed concurrently to confirm method accuracy, precision, and linearity.

L-isoleucine is structurally identical in molecular weight to L-leucine and shares near-identical chromatographic behavior under many standard HPLC conditions, making accurate differentiation and quantification of individual BCAAs particularly challenging in complex amino acid matrices. LC-MS/MS with MRM detection provides the compound-specific selectivity needed to unambiguously quantify L-isoleucine alongside its structural isomers, ensuring that BCAA ratios and individual amino acid label claims are accurately verified. This level of analytical rigor is essential for sports nutrition products where precise BCAA ratios are a key product differentiator and consumer expectation.

This test quantifies L-leucine, an essential branched-chain amino acid (BCAA) and the primary trigger of muscle protein synthesis via the mTOR pathway, in dietary supplements, protein powders, and raw materials using Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS). L-leucine is one of the most widely used amino acids in sports nutrition and recovery formulations, and accurate quantification is critical for label claim verification and ensuring consistent dosing across batches. Results are reported in mg per serving or mg per gram to support cGMP compliance.

A representative sample is weighed and extracted using an aqueous acidic solvent or protein precipitation with acetonitrile to isolate free L-leucine from the matrix. The clarified extract is injected onto a reversed-phase 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 L-leucine are monitored for quantification and identity confirmation. Quantification is performed against a multi-point calibration curve prepared from a certified L-leucine reference standard, with a stable isotope-labeled internal standard used to correct for matrix effects and ensure accurate recovery across sample types.

L-leucine shares near-identical molecular weight and chromatographic behavior with its structural isomers L-isoleucine and L-norleucine, making UV-based HPLC methods insufficient for unambiguous quantification in complex supplement matrices. LC-MS/MS in MRM mode provides the molecular selectivity required to distinguish L-leucine from its isomers and accurately quantify it at the levels present in high-dose amino acid and BCAA formulations, supporting reliable label claim substantiation and batch-to-batch consistency.

This test quantifies L-Lysine, an essential amino acid important for nutritional labeling and quality control, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). It is applicable to various matrices including raw materials, finished products, capsules, and powders. The method provides sensitive detection with quantification limits down to low microgram per gram levels, ensuring accurate measurement in complex formulations.

Samples are prepared by aqueous extraction followed by filtration to remove particulates. The extract is injected into an LC-MS/MS system equipped with a reverse-phase column, using multiple reaction monitoring (MRM) mode to detect L-Lysine transitions. Quantification is performed using a calibration curve constructed from certified L-Lysine reference standards, with an isotopically labeled internal standard to correct for matrix effects and instrument variability. Method accuracy is verified through duplicate injections, quality control samples, and spike recovery experiments.

Results are reported in mg/g (raw materials) or mg/serving (finished products). Testing confirms label claims, ensures raw material purity, and verifies batch-to-batch consistency in production.

This test quantifies L-methionine — an essential sulfur-containing amino acid that serves as the precursor to S-adenosylmethionine (SAMe), cysteine, and glutathione, and plays a central role in methylation reactions, liver detoxification, and antioxidant defense — in dietary supplements, protein blends, amino acid formulations, and raw materials using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Accurate quantification is important for label claim verification and for confirming the methionine contribution in complete protein and multi-amino acid formulations. LC-MS/MS provides the sensitivity and compound-specific selectivity needed to accurately quantify L-methionine in complex matrices. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved or hydrolyzed in a suitable aqueous solvent, with care taken to minimize oxidation of the sulfur-containing side chain during sample preparation. An isotopically labeled internal standard (e.g., ¹³C- or ²H-labeled L-methionine) is added prior to sample preparation to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase or HILIC LC-MS/MS, with detection by electrospray ionization (ESI) in positive ion mode using multiple reaction monitoring (MRM) transitions specific to L-methionine. Quantification is performed against a multi-point external calibration curve prepared from a certified L-methionine reference standard. Quality control samples at multiple concentration levels are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

L-methionine is an essential amino acid with a reactive sulfur-containing side chain that is susceptible to oxidation to methionine sulfoxide during processing and storage, potentially reducing bioavailability and potency. LC-MS/MS with MRM detection provides the compound-specific selectivity needed to specifically quantify intact L-methionine and distinguish it from its oxidized form and other sulfur-containing amino acids such as cysteine and homocysteine in complex matrices. This level of analytical rigor supports label claim accuracy, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay measures L-Norvaline, a branched-chain amino acid analog commonly used in nitric oxide–support and pre-workout formulations. LC-MS/MS provides high specificity and sensitivity for accurate quantification in both raw materials and finished products.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. L-Norvaline is detected in multiple reaction monitoring (MRM) mode and quantified using certified reference standards. Internal calibration and QC checks ensure precision, accuracy, and reproducibility across matrices.

Testing confirms label claims, ensures raw material purity, and verifies consistent dosing in sports nutrition formulations.

This assay measures L-Ornithine using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). The method provides high sensitivity and selectivity for accurate quantification in raw materials and finished formulations.

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

Testing verifies label claims, ensures purity, and supports batch-to-batch consistency across amino acid formulations.

This test confirms the botanical identity of lotus (Nelumbo nucifera) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic alkaloid and flavonoid fingerprint — including nuciferine and quercetin glycosides — of the sample is compared against a certified Nelumbo nucifera reference standard to confirm species authenticity and detect substitution with other aquatic plant materials or unrelated botanical powders. Results are reported as confirmed identity or non-conforming.

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

Lotus is an emerging supplement ingredient with limited commercial supply of authenticated reference material, making species-level identity verification particularly important for quality assurance. HPTLC identity testing based on the characteristic nuciferine alkaloid profile provides a practical and defensible confirmation of botanical authenticity, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This test quantifies L-phenylalanine — an essential aromatic amino acid that serves as a precursor to tyrosine, dopamine, norepinephrine, and epinephrine — in dietary supplements, amino acid blends, protein hydrolysates, and raw materials using High-Performance Liquid Chromatography (HPLC). Accurate quantification is critical for label claim verification, ensuring that the declared amount of this essential amino acid is present in the finished product. This test is also relevant for products bearing phenylalanine content disclosures required for individuals with phenylketonuria (PKU), a metabolic disorder in which phenylalanine cannot be properly metabolized. Results are reported as a percentage or in milligrams per serving.

A representative sample is accurately weighed and hydrolyzed under acidic conditions (e.g., 6N hydrochloric acid at elevated temperature) if present in a protein-bound form, or dissolved directly in a suitable buffer if in free amino acid form. The sample is derivatized using a pre-column or post-column reagent — such as o-phthalaldehyde (OPA), phenylisothiocyanate (PITC), or 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC) — to enable UV or fluorescence detection. Separation is performed by reversed-phase HPLC on a C18 column, and quantification is performed against a multi-point external calibration curve prepared from a certified L-phenylalanine reference standard. System suitability and quality control standards are run concurrently to confirm method accuracy and precision.

L-phenylalanine is an essential amino acid that cannot be synthesized by the human body and must be obtained through diet or supplementation, making accurate potency verification important for product efficacy and consumer safety. Critically, phenylalanine is a mandatory disclosure ingredient for individuals with PKU, and inaccurate labeling of phenylalanine content in food and supplement products poses a direct health risk to this population. HPLC with derivatization provides the sensitivity and specificity needed to accurately quantify L-phenylalanine in complex amino acid and protein matrices, supporting both label claim compliance and regulatory disclosure requirements under 21 CFR 101 and 111.

This test quantifies L-Proline, an important amino acid relevant for nutritional and quality assessment, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). It is applicable to various matrices including raw materials, powders, capsules, and finished products. The method provides sensitive detection with a reporting limit of 0.1 mg/kg, ensuring accurate measurement for quality control purposes.

Samples are prepared by extracting 0.5 g of material with 10 mL of 0.1% formic acid in water, followed by centrifugation and filtration. The extract is analyzed using LC-MS/MS with electrospray ionization in positive mode, monitoring L-Proline transitions in multiple reaction monitoring (MRM) mode. Quantification is performed using an external calibration curve constructed from certified L-Proline standards ranging from 0.1 to 100 mg/L. Quality control includes duplicate injections, analysis of spiked samples for recovery assessment, and periodic injection of QC standards to verify instrument performance.

Results are reported in mg/g (raw material) or mg/serving (finished product). Testing confirms label accuracy, verifies raw material purity, and supports consistent manufacturing quality.

This assay quantifies L-theanine, a naturally occurring amino acid found in green tea and commonly used in calming and nootropic supplements. Using HPLC, it verifies L-theanine content in capsules, powders, and beverages to confirm label claims and ensure consistent dosing.

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

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm active content and detect any degradation or underformulation.

This test quantifies L-threonine — an essential amino acid that plays a critical role in protein synthesis, immune globulin production, intestinal mucosal integrity, and collagen formation — in dietary supplements, protein blends, amino acid formulations, and raw materials using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). Accurate quantification is important for label claim verification in amino acid and protein supplement products, as well as for confirming the threonine contribution in complete protein and BCAA formulations. LC-MS/MS provides the sensitivity and compound-specific selectivity needed to accurately quantify L-threonine in complex amino acid matrices. Results are reported as a percentage or in milligrams per gram or per serving.

A representative sample is accurately weighed and dissolved or hydrolyzed in a suitable aqueous solvent. An isotopically labeled internal standard (e.g., ¹³C- or ²H-labeled L-threonine) is added prior to sample preparation to correct for matrix effects and recovery variability. The extract is filtered and analyzed by reversed-phase or HILIC LC-MS/MS, with detection by electrospray ionization (ESI) in positive ion mode using multiple reaction monitoring (MRM) transitions specific to L-threonine. Quantification is performed against a multi-point external calibration curve prepared from a certified L-threonine reference standard. Quality control samples at multiple concentration levels are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

L-threonine is an essential amino acid that cannot be synthesized by the human body, making accurate potency verification important for confirming that supplement products deliver the declared dose. LC-MS/MS with MRM detection provides the compound-specific selectivity needed to unambiguously quantify L-threonine in complex protein and amino acid matrices, where co-eluting amino acids may interfere with less selective analytical methods. This level of analytical rigor supports label claim accuracy, raw material qualification, and cGMP compliance under 21 CFR 111.

This test quantifies L-Tryptophan, an essential amino acid important for nutritional and quality control purposes, using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). It is suitable for analyzing raw materials, finished products, capsules, and powders. The method provides high specificity and sensitivity with detection limits typically in the low nanogram per milliliter range.

Samples are first extracted with a methanol-water solution to release L-Tryptophan, followed by filtration to remove particulates. The extract is injected into an LC-MS/MS system operating in Multiple Reaction Monitoring (MRM) mode, using electrospray ionization in positive mode. Separation is achieved on a reversed-phase C18 column with a gradient mobile phase of water and acetonitrile containing 0.1% formic acid. Quantification is performed using a calibration curve constructed from certified L-Tryptophan standards, with an isotopically labeled internal standard to correct for matrix effects and instrument variability. Method accuracy and precision are verified through duplicate injections, quality control samples, and spike recovery experiments.

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

This test quantifies L-Tyrosine, an important amino acid, in raw materials, powders, capsules, and finished products using High-Performance Liquid Chromatography (HPLC). Accurate measurement of L-Tyrosine ensures product quality and compliance with nutritional specifications. The assay has a detection limit of 0.1 mg/g, providing sensitive and precise quantification.

Samples are prepared by aqueous extraction followed by filtration to remove particulates. The extract is injected into an HPLC system equipped with a C18 reversed-phase column and UV detection at 274 nm, specific for L-Tyrosine. Quantification is achieved using an external calibration curve constructed from certified L-Tyrosine reference standards over a defined concentration range. Method precision is confirmed by duplicate injections and quality control samples, while spike recovery tests validate accuracy.

Results are reported in mg/g (raw material) or mg/serving (finished products). Testing confirms label claims, verifies raw material purity, and ensures batch-to-batch consistency in amino acid formulations.

This test quantifies the fibrinolytic activity of lumbrokinase — a complex of serine proteases derived from Lumbricus rubellus (red earthworm) with potent fibrin-degrading activity — in dietary supplements and raw materials, expressed in Fibrinolytic Units (FU) using the Standard Plasminogen Unit (SPU) activity assay method. Unlike mass-based quantification, this bioactivity assay directly measures the enzyme's functional capacity to degrade fibrin, providing a physiologically relevant measure of lumbrokinase potency. Accurate activity quantification is essential for label claim verification and for ensuring that the declared FU count reflects true enzymatic potency in the finished product. Results are reported in FU per gram or per serving.

A representative sample is dissolved in a suitable aqueous buffer at a defined pH and temperature to activate the lumbrokinase enzyme complex. The fibrinolytic activity is measured using a standardized substrate assay in which the sample is incubated with a fibrin or chromogenic substrate under controlled conditions. In the SPU method, the rate of fibrin degradation or chromogenic substrate hydrolysis is measured spectrophotometrically, and the activity is calculated by comparison to a certified lumbrokinase or plasminogen reference standard with a known FU activity. Assay conditions including pH, temperature, incubation time, and substrate concentration are tightly controlled throughout the analysis, and positive and negative controls are run concurrently to confirm assay validity and reproducibility.

Lumbrokinase is a high-value, activity-dependent enzyme ingredient where potency is defined by fibrinolytic activity rather than protein mass. Processing conditions, storage temperature, and formulation excipients can all affect enzymatic activity without changing the total protein content, making activity-based testing essential for confirming that the product delivers its intended physiological effect. The SPU fibrinolytic activity method provides a standardized, reproducible measure of lumbrokinase potency that is directly relevant to its mechanism of action, supporting label claim accuracy and consumer confidence in cardiovascular enzyme supplement products.

This assay quantifies lutein, a carotenoid found in marigold extract and leafy greens, commonly used in vision support and antioxidant supplements. Using HPLC, it measures lutein content in raw materials and finished products to confirm potency and support structure-function claims.

Samples are extracted using organic solvents (typically hexane or ethanol-based), then analyzed by HPLC with UV-Vis detection at a compound-specific wavelength (typically ~445 nm). Quantification is performed using certified lutein standards, with internal standard correction and duplicate injections for accuracy.

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

This assay quantifies luteolin, a bioactive flavonoid found in herbs, vegetables, and botanical supplements. Using LC-MS/MS, it measures luteolin content in capsules, powders, and plant extracts to verify label claims and ensure consistency in products targeting brain, immune, and inflammatory health.

Samples are extracted using alcohol- or methanol-based solvents, then analyzed by LC-MS/MS with compound-specific mass transitions. Quantification is performed using certified luteolin standards with internal standard correction and duplicate injections for precision and reproducibility.

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

This test quantifies L-valine — one of the three branched-chain amino acids (BCAAs), alongside leucine and isoleucine — in dietary supplements, protein blends, amino acid formulations, and raw materials using Liquid Chromatography coupled with Tandem Mass Spectrometry (LC-MS/MS). L-valine plays an important role in muscle tissue repair, nitrogen balance, and energy production during sustained exercise, and is a standard component of BCAA and sports nutrition formulations. LC-MS/MS provides the sensitivity and specificity required to accurately quantify L-valine and distinguish it from co-eluting structural isomers in complex amino acid matrices. Results are reported as a percentage or in milligrams per serving to support label claim verification and cGMP compliance.

A representative sample is accurately weighed and dissolved or hydrolyzed in a suitable aqueous solvent. An isotopically labeled internal standard (e.g., ¹³C- or ²H-labeled L-valine) is added prior to sample preparation to correct for matrix effects and recovery variability. The sample is filtered and injected onto a reversed-phase or HILIC LC column for chromatographic separation. Detection is performed by electrospray ionization (ESI) in positive ion mode, with multiple reaction monitoring (MRM) transitions selected to provide highly specific quantification of L-valine. Quantification is performed against a multi-point external calibration curve prepared from a certified L-valine reference standard. Quality control samples at multiple concentration levels are analyzed concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Accurate quantification of individual BCAAs — including L-valine — is essential for verifying the declared BCAA ratio and total amino acid content in sports nutrition products, where precise formulation is a key product claim and consumer expectation. LC-MS/MS with MRM detection provides the compound-specific selectivity needed to unambiguously quantify L-valine alongside leucine and isoleucine in complex protein and amino acid matrices, where standard HPLC methods may not provide sufficient resolution between structural isomers. This level of analytical specificity supports both raw material qualification and finished product release testing under 21 CFR 111 cGMP requirements.

This test quantifies lycopene, a fat-soluble carotenoid pigment found predominantly in tomatoes and tomato-derived ingredients, in dietary supplements, food products, and raw materials using High-Performance Liquid Chromatography with UV/Visible detection (HPLC-UV/Vis). Lycopene is one of the most studied carotenoids for its antioxidant activity and its association with prostate and cardiovascular health. 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 subjected to saponification using an ethanolic potassium hydroxide solution to hydrolyze esterified carotenoids, followed by liquid-liquid extraction with an organic solvent such as hexane or ethyl acetate to isolate lycopene. The extract is evaporated under nitrogen, reconstituted in mobile phase, and injected onto a reversed-phase C18 or C30 HPLC column — C30 columns are preferred for improved resolution of lycopene from structurally similar carotenoid isomers. Detection is performed by UV/Vis at approximately 472 nm, corresponding to the characteristic visible absorbance of lycopene. All sample preparation steps are conducted under amber or reduced-light conditions to prevent photodegradation, and quantification is performed against a multi-point external calibration curve prepared from a certified lycopene reference standard.

Lycopene is highly susceptible to oxidative and photodegradation, and its accurate quantification requires careful sample handling combined with a chromatographic method capable of resolving it from cis-isomers and co-present carotenoids such as beta-carotene and phytoene. HPLC with UV/Vis detection at 472 nm provides the sensitivity and selectivity needed for reliable potency measurement across softgel, powder, and oil-based supplement formats, ensuring label claim accuracy and raw material qualification.

This test confirms the botanical identity of maca root (Lepidium meyenii) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic glucosinolate and macamide fingerprint of the sample is compared against a certified Lepidium meyenii reference standard to confirm species authenticity and detect substitution with starch-based fillers or unrelated root powders. Results are reported as confirmed identity or non-conforming.

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

Maca root commands a significant price premium and is a known target for adulteration with cheaper root powders or starch fillers. HPTLC identity testing provides a defensible confirmation of botanical species, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This test confirms the botanical identity of maca root (Lepidium meyenii) in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). HPTLC generates a characteristic chromatographic fingerprint based on maca's unique glucosinolate and alkaloid marker compounds, which is compared against a certified Lepidium meyenii reference standard to confirm species authenticity and detect substitution or adulteration. Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol or an aqueous-organic solvent system and applied alongside a certified maca root reference standard onto an HPTLC silica gel plate using an automated applicator. The plate is developed in a validated solvent system, dried, and derivatized with an appropriate detection reagent to visualize the characteristic marker bands associated with maca's glucosinolate and macamide profile. The resulting fingerprint is compared visually and by densitometric scanning to the reference standard pattern. Identity is confirmed when the sample fingerprint matches the reference in terms of Rf values, band positions, and color profile, per established HPTLC botanical identity methods.

Maca root is a premium Andean botanical that commands a significant price premium, making it a target for substitution with less expensive root powders or unrelated starch-based fillers. HPTLC identity testing provides a rapid and scientifically defensible confirmation of botanical species, supporting supplier qualification, and label accuracy.

This assay quantifies malic acid, a naturally occurring organic acid found in fruits and commonly added as an acidulant in foods, beverages, and supplements. Using LC-MS/MS, it measures malic acid content to confirm formulation accuracy, detect adulteration, and verify nutrition or ingredient label claims.

Samples are extracted in aqueous solvent and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed with certified malic acid standards, using internal standard correction and duplicate injections to ensure sensitivity and reproducibility. The method allows detection of both natural and added malic acid in complex matrices.

Results are reported in mg per g, mg per 100 mL, or per serving depending on the product matrix. Values are compared against declared label claims, formulation targets, or regulatory specifications to ensure accuracy and product quality.

This assay measures the Dextrose Equivalent (DE) of maltodextrin using a titration method compliant with the USP monograph for maltodextrin. DE reflects the level of hydrolysis of starch into simpler sugars and is used to classify maltodextrin functionality and purity.

Samples are hydrolyzed under controlled acidic conditions to convert polysaccharides to reducing sugars. The concentration of reducing sugars is determined by titration with Fehling’s solution. The result is expressed as DE, which indicates the percentage of total reducing sugar content relative to dextrose.

Results are reported as Dextrose Equivalent (DE) units. USP guidelines typically specify DE ranges (e.g., 4–20) for maltodextrin depending on grade. This value helps confirm the degree of hydrolysis and ensures material matches the expected performance characteristics.

This assay quantifies maltodextrin content using a titration-based method in accordance with FCC (Food Chemicals Codex) specifications. It is used to verify maltodextrin identity and purity in powders and blends, especially in carrier systems or carbohydrate-rich functional products.

Samples are hydrolyzed under acidic conditions to release reducing sugars. The amount of reducing sugar is then measured by titration using Fehling’s solution or an equivalent FCC-validated method. Results are calculated as dextrose equivalents to assess maltodextrin concentration.

Results are reported as % w/w of maltodextrin or as dextrose equivalents (DE). This method helps confirm the identity, purity, and functionality of maltodextrin in food and supplement applications.

This assay quantifies mannitol, a sugar alcohol widely used as a low-calorie sweetener, bulking agent, and functional ingredient. Using LC-MS/MS, it measures mannitol levels in supplements, foods, and functional beverages to confirm label claims, verify formulation accuracy, and support “sugar-free” or “low-glycemic” positioning.

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

Results are reported in g per 100 g, g per 100 mL, or per serving. Values are compared with nutrition label claims and formulation specifications to confirm compliance, detect overages, and prevent undeclared sugar alcohols.

This test quantifies melanin — a class of high-molecular-weight, heterogeneous biopolymeric pigments produced through the oxidative polymerization of phenolic and indolic precursors — in botanical raw materials, fungal extracts (such as chaga mushroom), and dietary supplements using UV-Visible (UV-Vis) Spectrophotometry. Melanins are broadly classified into eumelanins (black-brown, indole-based), pheomelanins (yellow-red, sulfur-containing), and allomelanins (found in plants and fungi), and are valued in dietary supplement applications for their antioxidant properties and role as marker compounds for certain botanical and fungal ingredients. UV-Vis spectrophotometry exploits melanin's characteristic broad-spectrum light absorption, which increases continuously from the visible into the UV range, to provide a rapid and practical measure of total melanin content. Results are reported as a percentage or in milligrams per gram, typically expressed as melanin equivalents relative to a reference standard.

A representative sample is accurately weighed and extracted using an appropriate alkaline aqueous solution (e.g., dilute sodium hydroxide) to solubilize melanin polymers, which are poorly soluble under neutral or acidic conditions. The extract is filtered or centrifuged to remove insoluble particulates, and the absorbance of the clarified solution is measured spectrophotometrically at a characteristic wavelength — typically 400 nm or 475 nm — against a solvent blank. Total melanin concentration is calculated by comparison to a multi-point calibration curve prepared from a certified melanin reference standard (e.g., synthetic DOPA-melanin or Sepia melanin) or using the Beer-Lambert law with the established molar absorptivity for the reference melanin. All measurements are performed in triplicate and averaged for final quantification.

Melanin's broad, featureless UV-Vis absorption spectrum — arising from its extensively conjugated, cross-linked polymeric structure — makes UV-Vis spectrophotometry a practical and well-suited method for total melanin quantification. While UV-Vis does not resolve individual melanin subtypes or monomeric precursors, it provides a reliable measure of total melanin content appropriate for quality control of melanin-rich materials such as chaga mushroom extract, where melanin is a key marker compound and contributor to antioxidant activity. This method supports raw material potency verification, supplier qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies melatonin, a hormone commonly used in sleep-support supplements. Using HPLC, it verifies melatonin content in tablets, capsules, powders, and functional blends to ensure label accuracy and proper dosing in products designed for rest and relaxation.

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

Results are reported in mg per g or per serving. Values are compared with formulation targets and declared label claims to confirm potency, detect over- or under-formulation, and ensure product consistency.

This test quantifies methylene blue, a dye commonly used in pharmaceutical and food applications, to ensure product safety and compliance. Using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS), the method accurately measures methylene blue in various matrices such as solutions, powders, and finished formulations. The assay provides sensitive detection with quantification limits down to low microgram per liter levels.

Samples are first diluted in an aqueous solvent to extract methylene blue, followed by filtration to remove particulates. The prepared extracts are analyzed using LC-MS/MS with electrospray ionization in positive mode, monitoring methylene blue via multiple reaction monitoring (MRM) transitions specific to its molecular ions. Quantification is performed using a calibration curve constructed from certified methylene blue reference standards. Method accuracy and precision are verified through duplicate injections, matrix spike recoveries, and quality control samples analyzed alongside each batch.

Results are reported in µg/mL (liquids) or µg/g (solids). Testing verifies dosage accuracy, confirms product uniformity, and ensures compliance with quality and purity standards.

This assay quantifies methylliberine, the purine alkaloid branded as Dynamine, commonly used in energy, nootropic, and performance supplements. Using HPLC, it verifies methylliberine content to ensure label accuracy, consistent dosing, and safe stimulant formulation.

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 methylliberine 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 declared label claims to confirm potency and detect mislabeling or improper standardization.

This test identifies and quantifies methylsulfonylmethane (MSM), also known as dimethyl sulfone, in raw materials and finished dietary supplement products using liquid chromatography–tandem mass spectrometry (LC-MS/MS). MSM is a low-molecular-weight organosulfur compound commonly formulated in powders, capsules, tablets, gummies, liquids, and multi-ingredient joint- and mobility-support products. Because MSM is highly water-soluble and has limited chromophoric response for conventional UV detection, LC-MS/MS provides selective detection in the presence of amino acids, sugars, minerals, botanical extracts, flavor systems, and other formulation components. Results are reported as % w/w, mg/g, or mg per serving, according to the product specification and intended label declaration.

A representative sample is accurately weighed and dissolved or extracted in a validated aqueous or aqueous-organic solvent system, typically purified water or a water/methanol mixture, selected to achieve complete recovery of MSM from the relevant dosage form. Solid materials are mixed, sonicated, and/or agitated as necessary; finished matrices may undergo additional clarification steps to remove insoluble excipients and reduce matrix interference. The prepared extract is centrifuged and membrane-filtered before analysis. MSM is separated using a validated chromatographic mode suitable for highly polar analytes, such as hydrophilic-interaction liquid chromatography (HILIC) or a suitably retained reversed-phase method. Detection is performed by tandem mass spectrometry using optimized atmospheric-pressure ionization and compound-specific precursor/product-ion transitions. An isotopically labeled MSM internal standard or a qualified structurally appropriate internal standard may be used to correct for sample-preparation variability and matrix effects. Quantification is performed against a multi-point calibration curve prepared from a qualified, purity-corrected MSM reference standard. Method blanks, system-suitability standards, duplicate preparations, spike-recovery samples, calibration-verification standards, and concurrent quality-control samples are evaluated to confirm selectivity, accuracy, precision, linearity, recovery, and carryover control.

MSM is a small, polar, nonvolatile compound that may be challenging to retain and distinguish from formulation components using non-specific analytical approaches. LC-MS/MS offers high analyte specificity through chromatographic retention, molecular-ion detection, and characteristic fragmentation behavior, enabling reliable MSM quantification even in complex multi-ingredient supplements. The use of an internal standard and matrix-appropriate extraction conditions helps control potential ion-suppression and recovery effects that may otherwise influence results. This method supports raw-material qualification, formulation verification, lot-to-lot consistency, label-claim substantiation, and dietary supplement cGMP quality-control requirements under 21 CFR 111.

This assay quantifies methylsulfonylmethane (MSM), a sulfur-containing compound commonly used in joint health and recovery supplements. Using LC-MS/MS, it verifies MSM content in capsules, powders, and blends to confirm label accuracy and ensure consistent therapeutic dosing.

Samples are extracted in aqueous solution and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed with certified MSM 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 with formulation targets and label claims to confirm dosing consistency and detect dilution or mislabeling.

This test confirms the identity of milk thistle (Silybum marianum (L.) Gaertn.) in raw materials, seed powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Silybum marianum is one of the most extensively studied hepatoprotective botanicals, with its bioactivity attributed to the silymarin complex — a group of flavonolignan compounds including silybin A and B, isosilybin A and B, silychristin, silydianin, and taxifolin — concentrated in the seeds and fruit. Milk thistle is widely used in dietary supplements for liver health, detoxification support, and antioxidant protection. HPTLC identity testing generates a characteristic chromatographic fingerprint anchored by the silymarin flavonolignan profile that is compared against an authenticated S. marianum reference standard to confirm species identity and detect potential adulteration, substitution with other thistle species, or blending with inferior materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic flavonolignan profile of S. marianum. The extract is applied alongside a certified milk thistle 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 silymarin flavonolignan constituents of S. marianum. After development, the plate is derivatized with Natural Products Reagent A (NP/PEG) — the preferred reagent for flavonoid and flavonolignan visualization — and evaluated under UV light at 366 nm, where the silymarin components display characteristic orange-yellow fluorescence. The plate is also evaluated under UV light at 254 nm and under white light after 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.

Milk thistle raw materials are subject to adulteration and species substitution, including replacement with other thistle species (e.g., Carduus spp., Cirsium spp.) that lack the characteristic silymarin flavonolignan profile and associated hepatoprotective activity. HPTLC fingerprinting with NP/PEG derivatization provides a holistic, multi-compound chromatographic identity confirmation that is anchored by the distinctive fluorescent silymarin band pattern, enabling detection of substitution or adulteration that would not be apparent from single-marker silybin assays alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This assay quantifies the key active flavonolignans in milk thistle (Silybum marianum), commonly grouped under the silymarin complex. Using LC-MS/MS, it measures individual and total silymarin content to verify potency in liver support supplements and botanical blends.

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

Results are reported in mg per g or per serving for each analyte and total silymarin. Values are compared to formulation targets and label claims to confirm extract standardization and detect potential degradation or adulteration.

This assay quantifies total mogrosides, the active sweet compounds found in monk fruit (Siraitia grosvenorii). Using LC-MS/MS, it measures mogroside content—typically focusing on mogroside V and related glycosides—to confirm natural sweetener potency, label accuracy, and protection against added sugar adulteration.

Samples are extracted using aqueous or alcohol-based solvents and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified mogroside 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 with formulation targets and label claims to verify sweetening strength and ensure consistency across monk fruit-based formulations.

This assay quantifies mogroside V, the primary sweet compound in monk fruit (Siraitia grosvenorii). Using LC-MS/MS, it verifies mogroside V content in natural sweeteners, beverages, and functional formulations to ensure accurate label claims and consistent sweetness in low- or no-sugar products.

Samples are extracted using aqueous or alcohol-based solvents, then analyzed by LC-MS/MS with compound-specific mass transitions. Quantification is performed using certified mogroside V 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 sweetness potency, detect dilution, or identify non-monk fruit adulterants.

This assay confirms the botanical identity of Monascus purpurea using High-Performance Thin Layer Chromatography (HPTLC). Characteristic marker compounds and chromatographic fingerprints are compared against authentic reference standards.

Samples are extracted and applied to an HPTLC plate alongside reference materials. After chromatographic development, plates are visualized under specific light and/or derivatization reagents. The resulting banding pattern is compared to authentic Monascus purpurea for positive identification.

Results are reported qualitatively as “Conforms” (identity verified) or “Does Not Conform.” Testing ensures raw material authenticity, detects adulteration or substitution, and supports regulatory and quality compliance.

This assay measures monolaurin (1-lauroyl-rac-glycerol), a glycerol monoester of lauric acid commonly used for immune and microbiome support. LC-MS/MS provides high sensitivity and selectivity for distinguishing monolaurin from other medium-chain glycerides and fatty acids.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Monolaurin is detected in multiple reaction monitoring (MRM) mode and quantified using certified reference standards. Internal calibration and QC checks ensure reproducible and accurate results across lipid-rich matrices.

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

This test confirms the identity of moringa (Moringa oleifera) in raw materials, leaf powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Moringa oleifera, commonly known as the drumstick tree or miracle tree, is a tropical plant widely used in dietary supplements for its exceptional nutritional density and content of bioactive compounds including isothiocyanates, glucosinolates, flavonoids (notably quercetin and kaempferol glycosides), and chlorogenic acid derivatives. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated M. oleifera reference standard to confirm species identity and detect potential adulteration, substitution with other Moringa species, or blending with inferior plant materials.

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

Moringa raw materials are subject to adulteration and species substitution, including blending with other Moringa species (e.g., M. stenopetala) or unrelated plant powders, particularly given the growing global demand for moringa as a superfood ingredient. 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 nutritional or 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 assay quantifies L-DOPA, the primary bioactive compound in Mucuna pruriens (velvet bean), using LC-MS/MS. It verifies the potency and standardization of Mucuna extracts in supplements formulated for neurological support, motivation, and men’s health.

Samples are extracted using acidified aqueous solvents and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed using certified L-DOPA 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 to standardization targets (e.g., 15% or 20% L-DOPA) and label claims to confirm potency, consistency, and raw material quality.

This assay quantifies N-acetylcysteine (NAC), a stable derivative of the amino acid cysteine widely used in supplements for antioxidant support and glutathione production. Using HPLC, it measures NAC content in raw materials and finished products to confirm label claims, ensure potency, and detect degradation.

Samples are extracted in aqueous or methanolic solution and analyzed by HPLC with UV detection at a compound-specific wavelength (typically ~210 nm). Quantification is performed using certified NAC 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 with declared label claims and formulation targets to verify potency, support stability checks, and confirm compliance with product specifications.

This test quantifies N-Acetyl-D-Glucosamine (GlcNAc), an important amino sugar involved in the structure of glycoproteins and chitin, using LC-MS/MS. It is applicable to various sample types including raw materials, finished products, capsules, and powders. The method offers sensitive detection with a reporting limit of 0.1 mg/kg, ensuring accurate measurement even in complex matrices.

Samples are prepared by extracting with aqueous methanol followed by filtration to remove particulates. Analysis is performed using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) operating in multiple reaction monitoring (MRM) mode, targeting specific GlcNAc transitions. Quantification is achieved through an external calibration curve constructed from certified GlcNAc reference standards. Internal standards are used to correct for matrix effects and instrument variability. Method accuracy and precision are confirmed by analyzing quality control samples and performing spike recovery tests, with duplicate injections conducted for each sample.

Results are reported in mg/g (raw material) or mg/serving (finished products). Testing verifies ingredient purity, supports formulation quality, and ensures compliance with product specifications.

This test quantifies N-Acetyl-L-Carnitine (ALCAR), a compound important for cellular energy metabolism and neurological function, using LC-MS/MS. It is applicable to raw materials, capsules, powders, and finished dietary supplements. The method offers sensitive detection with a reporting limit of 0.1 mg/kg, ensuring accurate measurement for quality control and regulatory compliance.

Samples are prepared by extracting 0.5 g of material with 10 mL of methanol-water (80:20 v/v) followed by centrifugation and filtration. The extract is analyzed using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with electrospray ionization in positive mode, monitoring specific MRM transitions for ALCAR. Quantification is performed using a calibration curve generated from certified ALCAR reference standards ranging from 0.1 to 50 mg/L. Method accuracy and precision are verified through duplicate injections, spiked recovery samples, and quality control standards analyzed alongside each batch.

Testing confirms label accuracy, verifies purity, and supports consistency in products formulated with carnitine derivatives.

This assay quantifies N-Acetyl-L-Cysteine Ethyl Ester (NACET), a highly bioavailable derivative of NAC, in supplement and functional ingredient products. Using HPLC, it verifies NACET content to ensure correct dosing, support label claims, and confirm product integrity in advanced antioxidant and detox formulations.

Samples are extracted under stabilized, pH-controlled conditions to preserve the NACET structure. The extract is analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is achieved using high-purity NACET standards, with internal standard correction and duplicate runs to ensure reliable results.

Results are reported in mg per g or per serving. Values are compared with formulation targets and label claims. Testing confirms proper NACET delivery and helps distinguish premium NACET-based products from standard NAC or mislabeled formulations.

This assay quantifies N-acetyl-L-tyrosine (NALT), an acetylated derivative of the amino acid tyrosine used in nootropic and performance supplements. Using HPLC, it measures NALT content in raw materials and finished products to verify potency, ensure label accuracy, and confirm consistency across production batches.

Samples are extracted in aqueous or methanolic solution and analyzed by HPLC with UV detection at a compound-specific wavelength (typically ~274 nm for aromatic amino acids). Quantification is performed using certified N-acetyl-L-tyrosine reference standards, with internal standard correction and duplicate injections to ensure reproducibility and accuracy.

Results are reported in mg per g or per serving. Values are compared against declared label claims and formulation specifications to confirm dosing accuracy, detect degradation, or identify adulteration.

This test quantifies naringin, a flavanone glycoside and the primary bitter bioactive compound found in grapefruit and other citrus fruits, in dietary supplements, citrus extracts, and raw materials using High-Performance Liquid Chromatography with UV detection (HPLC-UV). Naringin is widely used as a potency marker for grapefruit and citrus bioflavonoid extracts and has been studied for its antioxidant, anti-inflammatory, and lipid-modulating 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 or gentle heating to ensure complete solubilization of naringin 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 283–284 nm, and quantification is performed against a multi-point external calibration curve prepared from a certified naringin reference standard. System suitability and QC samples are run concurrently to confirm method accuracy and reproducibility across the analytical run.

Naringin is the primary standardization marker for grapefruit extract and citrus bioflavonoid ingredients, and accurate quantification is essential for verifying extract potency and label claim compliance. HPLC-UV provides the selectivity needed to resolve naringin from structurally related flavonoids — including hesperidin, narirutin, and neohesperidin — that are commonly co-present in citrus-derived matrices, delivering reliable potency data for both raw material qualification and finished product release testing.

This test confirms the identity of neem (Azadirachta indica A. Juss.) in raw materials, leaf, bark, or seed powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Azadirachta indica, commonly known as neem or Indian lilac, is one of the most extensively used medicinal plants in Ayurvedic and traditional medicine, valued for its broad-spectrum biological activities including antimicrobial, anti-inflammatory, antioxidant, and immunomodulatory properties. Its characteristic phytochemical profile includes limonoids (azadirachtin, nimbin, nimbidin, salannin, gedunin), flavonoids (quercetin, kaempferol), tannins, and terpenoids. HPTLC identity testing generates a characteristic chromatographic fingerprint — anchored by the distinctive limonoid profile — that is compared against an authenticated A. indica reference standard to confirm species identity and detect potential adulteration, substitution with other Meliaceae family members, or blending with unrelated plant materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol, ethanol, or a combination of non-polar and polar solvents) to capture the characteristic secondary metabolite profile of A. indica, including limonoids, flavonoids, and tannins. The extract is applied alongside a certified neem 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 limonoid and flavonoid constituents of A. indica. After development, the plate is derivatized with an appropriate reagent (e.g., anisaldehyde-sulfuric acid or vanillin-sulfuric acid for limonoid visualization, or Natural Products Reagent A / NP/PEG for flavonoid visualization) 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.

Neem raw materials are subject to adulteration and species substitution, including replacement with other Meliaceae family members or unrelated plant materials that may share morphological similarities in dried and powdered form. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation — anchored by the characteristic limonoid band pattern unique to A. indica — that is more discriminating than single-marker assays, enabling detection of substitution or adulteration that would not be apparent from potency testing alone. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This test confirms the identity of nettle leaf (Urtica dioica L., and related species including U. urens) in raw materials, dried leaf powders, extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Urtica dioica, commonly known as stinging nettle, is a widely used medicinal and nutritional herb with a long history of use in traditional European and Ayurvedic medicine for supporting joint and musculoskeletal health, allergic rhinitis, benign prostatic hyperplasia (BPH), and urinary tract function. Its characteristic phytochemical profile includes flavonoids (quercetin, kaempferol, isorhamnetin, and their glycosides including rutin and isoquercitrin), hydroxycinnamic acids (chlorogenic acid, caffeic acid), lectins (Urtica dioica agglutinin, UDA), sterols (β-sitosterol, stigmasterol), and polysaccharides. HPTLC identity testing generates a characteristic chromatographic fingerprint that is compared against an authenticated U. dioica reference standard to confirm species identity and detect potential adulteration, substitution with other Urtica species or unrelated plant materials, or blending with inferior leaf materials.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic flavonoid and phenolic acid profile of U. dioica leaf. The extract is applied alongside a certified nettle leaf 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 flavonoid glycosides and phenolic acids of U. dioica, including rutin and chlorogenic acid as key marker compounds. After development, the plate is derivatized with Natural Products Reagent A (NP/PEG) — the preferred reagent for flavonoid visualization — and evaluated under UV light at 366 nm, where the characteristic flavonoid bands display orange-yellow fluorescence. The plate is also evaluated under UV light at 254 nm and under white light after 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.

Nettle leaf raw materials are subject to adulteration and species substitution, including replacement with other Urtica species, related Urticaceae family members, or unrelated leaf materials that may share morphological similarities in dried and powdered form. HPTLC fingerprinting with NP/PEG derivatization provides a holistic, multi-compound chromatographic identity confirmation — anchored by the characteristic flavonoid glycoside pattern, particularly the rutin and isoquercitrin bands — 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 assay quantifies NAD⁺ (nicotinamide adenine dinucleotide), a vital coenzyme involved in redox reactions and mitochondrial energy metabolism. Using HPLC, it verifies NAD⁺ content in supplements and functional products to confirm bioactive potency and support claims related to energy, longevity, and cellular health.

Samples are extracted under cold, acid-stabilized conditions to preserve NAD⁺ integrity, then analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified NAD⁺ 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 degradation, which NAD⁺ is particularly susceptible to in heat or pH-variable environments.

This assay quantifies nicotinamide riboside, a bioavailable form of vitamin B3 that acts as a precursor to NAD⁺. The test measures active content using LC-MS/MS to ensure potency and formulation accuracy in supplements targeting metabolic and cellular health.

Samples are extracted under controlled conditions to preserve nicotinamide riboside. 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. Stable levels across batches confirm formulation consistency, while deviations may indicate instability or degradation.

This assay quantifies nicotine levels in food or plant-derived products using high-performance liquid chromatography (HPLC). Nicotine is an alkaloid commonly found in tobacco and related species, and its measurement is critical in ensuring compliance with regulatory standards and validating product labeling. The method uses a validated HPLC protocol that isolates nicotine from complex matrices, achieving accurate quantification with minimal interference.

Samples are homogenized and subjected to solvent extraction using a method optimized for nicotine recovery. After filtration and cleanup, the extract is analyzed by HPLC, which separates nicotine from other matrix components based on retention time. Quantification is performed using UV detection against calibration curves prepared with qualified reference standards. Quality controls and internal standards ensure reproducibility and accuracy.

Results are reported as numerical concentrations (typically in ppm or µg/g). Lower values indicate minimal nicotine presence, often consistent with regulatory thresholds for non-tobacco products. The results support both compliance verification and quality control for product consistency across batches.

This assay quantifies nitrate content in beet juice, powders, and related raw materials using Ion Chromatography (IC). It measures nitrate ions directly, ensuring accurate assessment of beet products marketed for cardiovascular support, endurance, and nitric oxide boosting effects.

Samples are dissolved in water and filtered before analysis by ion chromatography with conductivity detection. Nitrate ions are separated on an anion-exchange column and quantified against certified nitrate standards. Duplicate injections and QC samples are run to ensure reproducibility and accuracy.

Results are reported in mg/g (powders) or mg/100 mL (juices), and may also be expressed per serving. Values are compared against product specifications and label claims to confirm potency and detect batch-to-batch variability.

This assay measures NMNH (dihydronicotinamide mononucleotide), the reduced form of NMN, using High-Performance Liquid Chromatography (HPLC). The analysis provides accurate quantification for quality control and standardization in raw materials and finished products.

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

Testing verifies standardized potency, confirms label claims, and ensures batch-to-batch consistency in NMNH-containing formulations.

This assay quantifies NMN (Nicotinamide Mononucleotide), a precursor to NAD⁺ widely used in longevity and mitochondrial health supplements. Using HPLC, it verifies NMN content in raw materials and finished products to ensure label accuracy, proper dosing, and product stability.

Samples are extracted in aqueous or buffered solution and analyzed by HPLC with UV detection at a compound-specific wavelength (typically ~260 nm). Quantification is performed using certified NMN standards with internal standard correction and duplicate injections to ensure precision and

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

This comprehensive assay compiles a full nutritional profile of a food product by integrating results from multiple individual tests. It measures macronutrients, fiber, vitamins, minerals, and other components to provide the complete data required for label statements and nutritional information.

The sample is subjected to a series of standardized assays (e.g., Dumas for protein, Soxhlet for fat, AOAC methods for fiber, HPLC for vitamins) under controlled conditions. Data from each test are compiled and cross-checked against quality control samples. The final nutritional facts panel is reviewed for accuracy before reporting.

Results are delivered as quantitative values (e.g., calories, grams per serving) for each nutrient. These data enable manufacturers to verify that their products meet nutritional targets and comply with labeling requirements. The integrated panel provides a clear overview of the product’s nutritional composition.

This assay quantifies oleuropein, a polyphenolic compound found in olive leaves and fruit. Using HPLC, it verifies oleuropein content in botanical extracts, capsules, and functional blends to ensure accurate dosing and support claims related to heart health, inflammation balance, and immune function.

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

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm active content and assess extract quality or degradation.

This assay measures characteristic olive marker compounds using Liquid Chromatography–Tandem Mass Spectrometry (LC-MS/MS). Analysis typically targets olive polyphenols and secoiridoids to confirm ingredient identity and standardized active content.

Samples are extracted and analyzed under validated LC-MS/MS chromatographic conditions. Target olive markers 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 authenticity, supports label claims, and ensures batch-to-batch consistency in olive-derived formulations.

This assay determines total Oxygen Radical Absorbance Capacity (ORAC) by measuring both hydrophilic (water-soluble) and hydrophobic (fat-soluble) antioxidant fractions. The combined result reflects overall antioxidant activity across diverse compound classes.

Samples are separated into hydrophilic and hydrophobic fractions and analyzed using a UV-Vis–based ORAC assay. Antioxidant capacity is determined by monitoring the inhibition of fluorescence decay caused by peroxyl radicals, relative to a Trolox standard. Individual fractions are calculated and summed to yield Total ORAC.

Testing supports product comparison, standardization, and verification of antioxidant-related claims.

This assay measures PABA (para-aminobenzoic acid), a B-complex–related compound used in nutritional and cosmetic formulations. HPLC provides accurate and selective quantification of PABA in raw materials and finished products.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. PABA is separated from related aromatic compounds and detected via UV absorbance. Quantitation is performed with certified reference standards; calibration curves and replicate injections ensure accuracy and reproducibility.

Testing confirms label claims, verifies purity, and ensures consistency across production batches.

This assay measures palmitoylethanolamide (PEA), also known as palmitic acid monoethanolamide, using High-Performance Liquid Chromatography (HPLC). The analysis provides accurate quantification for quality control and standardization in raw materials and finished products.

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

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

This test confirms the botanical identity of Panax ginseng root in raw materials and finished products using High-Performance Thin-Layer Chromatography (HPTLC). The characteristic ginsenoside fingerprint of the sample is compared against a certified Panax ginseng reference standard to confirm species authenticity and detect substitution with related species such as American ginseng (P. quinquefolius) or Siberian ginseng (Eleutherococcus senticosus). Results are reported as confirmed identity or non-conforming.

A representative sample is extracted using methanol and applied alongside a certified Panax ginseng 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 resulting fingerprint is compared visually and by densitometric scanning to the reference standard in terms of Rf values, band positions, and color profile.

Panax ginseng is one of the most frequently adulterated botanical ingredients globally, with substitution by less expensive ginseng species a well-documented industry problem. HPTLC identity testing provides a rapid and defensible confirmation of botanical species, supporting supplier qualification and cGMP compliance under 21 CFR 111.

This assay measures pantothenic acid originating from pantethine, a biologically active dimeric form of vitamin B5. HPLC analysis provides accurate quantification of pantethine-derived pantothenic acid in raw materials and finished products, ensuring correct dosing and formulation integrity.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. Pantethine is cleaved to yield measurable pantothenic acid, which is separated from other B-vitamin compounds and detected via UV or diode-array detection. Quantitation is performed using certified reference standards, with calibration curves and replicate injections ensuring precision and reproducibility.

Testing verifies label claims, confirms purity, and supports consistent manufacturing for pantethine-containing formulations.

This assay measures the enzymatic activity of papain, a cysteine protease derived from papaya latex, using the FCC (Food Chemicals Codex) method. Activity is reported in Papain Units (PU), which reflect the enzyme’s ability to hydrolyze protein substrates under standardized conditions.

Samples are incubated with a casein substrate under defined pH and temperature conditions. The extent of protein hydrolysis is measured by the release of soluble peptides, typically monitored spectrophotometrically after precipitation of unreacted protein. Activity is calculated against the FCC reference definition of one PU. Duplicate runs and standard curve validation ensure accuracy.

Results are reported in PU per gram or PU per serving. Values are compared to product specifications and label claims to verify enzyme strength, confirm stability, and ensure consistency across batches.

This assay quantifies paraxanthine, a metabolite of caffeine known for its clean stimulant effect with reduced side effects. Using HPLC, it verifies paraxanthine content in energy, nootropic, and pre-workout supplements to ensure proper dosing and label accuracy.

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

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

This test confirms the identity of partially hydrolyzed guar gum (PHGG) — a water-soluble, low-viscosity dietary fiber derived from the enzymatic hydrolysis of guar gum (Cyamopsis tetragonoloba (L.) Taub.) galactomannan — in raw materials and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). PHGG is produced by controlled enzymatic hydrolysis of native guar gum, reducing its molecular weight and viscosity while preserving its galactomannan backbone structure composed of a (1→4)-β-D-mannose main chain with (1→6)-α-D-galactose side chains. PHGG is used in dietary supplements and functional foods as a prebiotic soluble fiber with clinical evidence supporting improvements in gut microbiota composition, bowel regularity, and digestive comfort. HPTLC identity testing employs acid hydrolysis to release the characteristic monosaccharide constituents — primarily mannose and galactose in the approximately 2:1 molar ratio characteristic of guar galactomannan — which are resolved and visualized as a diagnostic sugar fingerprint compared against an authenticated PHGG reference standard to confirm identity and detect potential adulteration or substitution with other galactomannan- or polysaccharide-containing materials.

A representative sample is accurately weighed and subjected to complete acid hydrolysis (e.g., using dilute sulfuric or hydrochloric acid under reflux conditions) to hydrolyze the galactomannan polysaccharide into its constituent monosaccharides. The hydrolysate is neutralized, filtered, and applied alongside an authenticated PHGG hydrolysate reference standard, individual monosaccharide reference standards (mannose, galactose), and, where applicable, potential adulterant hydrolysates (e.g., locust bean gum, fenugreek galactomannan), onto an HPTLC silica gel plate using an automated sample applicator. The plate is developed in a validated aqueous-organic solvent system optimized for monosaccharide resolution. After development, the plate is derivatized with an appropriate sugar-visualizing reagent (e.g., diphenylamine-aniline-phosphoric acid or anisaldehyde-sulfuric acid) and evaluated under white light and UV light. The resulting monosaccharide fingerprint — characterized by the characteristic mannose and galactose band pattern in the approximately 2:1 molar ratio diagnostic of guar galactomannan — is compared visually and, where applicable, by densitometric analysis to the authenticated reference standard in accordance with established HPTLC identity testing guidelines.

PHGG is subject to adulteration and substitution with other galactomannan-containing materials — including locust bean gum, fenugreek galactomannan, and native guar gum — as well as other polysaccharide fillers that may be visually indistinguishable in dried powder form. HPTLC monosaccharide fingerprinting following acid hydrolysis provides a practical and discriminating identity confirmation method that exploits the characteristic mannose-to-galactose ratio of guar galactomannan (approximately 2:1) to distinguish PHGG from other galactomannans with different monosaccharide ratios (e.g., locust bean gum ~4:1, fenugreek galactomannan ~1:1) and from non-galactomannan polysaccharide 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 verifies the identity of Passiflora incarnata using High-Performance Thin-Layer Chromatography (HPTLC). Characteristic chromatographic fingerprints are compared against authenticated reference material to confirm botanical identity.

Samples are extracted and applied to an HPTLC plate alongside reference standards. After chromatographic development and visualization under appropriate conditions, the resulting banding pattern is evaluated and compared to the reference profile for identity confirmation.

Testing supports raw material authentication, supplier verification, and quality control for botanical ingredients.

Initially, a sample is measured for the Amino Acid composition using method AOAC 994.12. A human digestion simulation follows, breaking down the proteins into amino acids, which are reacted with Ninhydrin and measured; determining the digestibility. Taking the limiting amino acid value the digestibility is corrected resulting in a Protein Digestibility Corrected Amino Acid Score (PDCAAS).

The sample undergoes controlled enzymatic digestion using the patented in vitro method. The extent of protein breakdown is measured and used to calculate PDCAAS.

Results are reported on a 0–1 scale. A score of 1.0 indicates high-quality protein, while lower scores suggest amino acid limitations or reduced digestibility.

This test confirms the botanical identity of peppermint raw materials — including dried leaf, leaf powder, and leaf extract — using High-Performance Thin-Layer Chromatography (HPTLC). Peppermint is derived from Mentha × piperita L. (Lamiaceae), a sterile hybrid of watermint (Mentha aquatica) and spearmint (Mentha spicata), recognized as an official article in major pharmacopoeias including the United States Pharmacopeia (USP), European Pharmacopoeia (Ph. Eur.), and British Pharmacopoeia (BP). Peppermint is widely used in dietary supplements for its digestive, carminative, antispasmodic, and cooling properties, with menthol, menthone, menthyl acetate, and rosmarinic acid serving as the principal marker compounds. The monoterpene composition of the essential oil — dominated by menthol and menthone — is characteristic of M. × piperita and distinguishes it from other commercially important mint species, particularly spearmint (M. spicata, characterized by carvone) and cornmint (M. arvensis, characterized by high menthol with low menthyl acetate). HPTLC identity testing generates a characteristic chromatographic fingerprint of the peppermint extract — encompassing both its volatile terpenoid and non-volatile phenolic acid profiles — that is compared against an authenticated reference standard to confirm species identity and detect potential substitution or adulteration.

A representative sample of peppermint raw material is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or 70% ethanol) to produce a total extract enriched in both the characteristic phenolic acid constituents (rosmarinic acid, caffeic acid) and, where applicable, the non-volatile terpenoid residues. For comprehensive volatile marker profiling, an additional extraction using a non-polar solvent (e.g., dichloromethane or hexane) may be employed to capture menthol, menthone, and menthyl acetate. The extract is applied alongside a certified Mentha × piperita reference standard and, where applicable, reference standards for menthol, menthone, and rosmarinic acid, onto a silica gel 60 F₂₅₄ HPTLC plate using an automated sample applicator to ensure precise, reproducible band placement. The plate is developed in a validated solvent system (e.g., toluene/ethyl acetate or ethyl acetate/formic acid/glacial acetic acid/water) optimized for resolution of peppermint's characteristic terpenoid and phenolic markers. After development, the plate is dried and visualized under UV light at 254 nm and 366 nm, followed by derivatization with an appropriate spray reagent (e.g., anisaldehyde-sulfuric acid or Natural Products Reagent A) and heating to reveal the full characteristic fingerprint. The resulting chromatographic pattern — including the positions, colors, and relative intensities of all characteristic zones — is compared with the authenticated reference standard. Identity is confirmed when the sample fingerprint is concordant with the reference. The method is performed in accordance with USP General Chapter <203> (HPTLC) and/or AHPA/AHP HPTLC reference standards.

HPTLC is the preferred method for botanical identity testing of peppermint because it generates a holistic chromatographic fingerprint encompassing both the terpenoid and phenolic acid profiles that together constitute the authentic chemical signature of Mentha × piperita. This multi-marker fingerprint approach is more discriminating than single-marker testing and is capable of distinguishing authentic peppermint from closely related and commercially significant mint species — including spearmint (M. spicata) and cornmint (M. arvensis) — that are common adulterants or substitutes in the botanical ingredient supply chain and would not be reliably differentiated by visual or organoleptic inspection alone. The characteristic terpenoid composition of peppermint's essential oil, particularly the ratio of menthol to menthone and the presence of menthyl acetate, provides a highly distinctive chemical identity marker that is faithfully captured in the HPTLC fingerprint. Botanical identity verification is a mandatory requirement under 21 CFR 111.75 for dietary supplement raw material qualification, and HPTLC identity testing is explicitly recognized as an appropriate identity testing methodology in USP General Chapter <203>.

This test quantifies phosphatidylcholine (PC), an essential phospholipid found in lecithin and cell membranes, using High-Performance Liquid Chromatography (HPLC). It is applicable to raw materials, finished products, capsules, and powders, providing precise measurement of PC content to ensure product quality and label accuracy. The method achieves detection limits suitable for trace-level quantification, with results reported in mg/g or percentage of total lipid content.

Samples are prepared by extracting lipids using a chloroform-methanol solvent mixture (2:1 v/v) followed by phase separation to isolate the lipid fraction. The extracted lipids are then injected into an HPLC system equipped with a C18 reverse-phase column. Phosphatidylcholine is detected using evaporative light scattering detection (ELSD) to enhance sensitivity for non-UV absorbing lipids. Quantification is performed by comparing peak areas to a calibration curve constructed from certified phosphatidylcholine reference standards. Method accuracy is verified through duplicate injections, inclusion of quality control samples, and spike recovery tests to confirm extraction efficiency and instrument precision.

Results are reported in % w/w (raw materials) or mg/serving (finished products). Testing confirms standardized potency, supports label claims, and ensures consistency of lecithin and lipid-based supplements.

This test quantifies phosphatidylserine (PS) — a phospholipid and critical structural component of neuronal cell membranes, widely used in dietary supplements for its FDA-qualified health claim supporting cognitive function and dementia risk reduction — in raw materials, lecithin-derived extracts, and finished products using High-Performance Liquid Chromatography (HPLC). Phosphatidylserine is typically derived from soy or sunflower lecithin and is standardized to a defined PS content (commonly 20% or higher). Accurate quantification is essential for verifying label claims, confirming the potency of standardized PS concentrates, and ensuring that the declared amount of this functional phospholipid 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 extracted using an appropriate lipid extraction solvent system (e.g., chloroform/methanol or isopropanol) to isolate the phospholipid fraction. The extract is filtered and analyzed by HPLC using a normal-phase or HILIC column with evaporative light scattering detection (ELSD) or charged aerosol detection (CAD), which are preferred over UV detection due to the absence of a strong UV chromophore in the phosphatidylserine headgroup. Where UV detection is employed, a wavelength of 205 nm may be used to detect the unsaturated fatty acid moieties. Quantification is performed against a multi-point external calibration curve prepared from a certified phosphatidylserine reference standard. Quality control samples are run concurrently to confirm method accuracy, precision, and linearity throughout the analytical run.

Phosphatidylserine is a complex phospholipid that co-occurs with other phospholipid classes — including phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol — in lecithin-derived raw materials. HPLC with ELSD or CAD detection provides the chromatographic resolution needed to separate and specifically quantify the PS fraction from co-occurring phospholipids without the need for mass spectrometric detection, making it a practical and cost-effective method for routine quality control. This approach supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111, and is consistent with the analytical requirements for products bearing FDA-qualified health claims for phosphatidylserine.

This test identifies and quantifies phospholipids in raw materials and finished dietary supplement products using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Depending on the validated assay scope and product matrix, the panel may include phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositol (PI), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidic acid (PA), lysophospholipids, and relevant sphingolipids. Individual molecular species may also be reported according to acyl-chain composition, such as PC 16:0/18:2 or PS 18:0/22:6. The assay is particularly suited to complex lipid-containing ingredients, including soy or sunflower lecithin, egg-derived phospholipids, krill oil, marine oils, milk-derived fractions, and phospholipid-enriched blends. Results can be reported as mg/g, mg/serving, % w/w, individual phospholipid-class concentration, individual molecular-species concentration, and/or total phospholipids calculated as the sum of validated phospholipid classes.

A representative sample is accurately weighed and subjected to a validated lipid-extraction procedure using an appropriate biphasic organic-solvent system, such as a modified Folch, Bligh–Dyer, or methyl tert-butyl ether extraction. Class-appropriate stable-isotope-labeled or structurally analogous internal standards are added before extraction to monitor recovery, ionization effects, and analytical variability. The lipid extract is concentrated, reconstituted in an LC-MS-compatible solvent, and analyzed by reversed-phase or hydrophilic-interaction liquid chromatography (HILIC), selected according to whether the validated method emphasizes separation by molecular species or phospholipid class. Tandem mass spectrometry is performed using validated electrospray-ionization conditions and compound-specific multiple-reaction-monitoring (MRM) or high-resolution MS/MS transitions. Analytes are identified by retention-time agreement, precursor-ion mass, characteristic fragment ions, and comparison with qualified reference standards. Quantification is based on multi-point calibration curves using certified or qualified phospholipid standards, with response correction by the corresponding internal standard where applicable. Method blanks, duplicate preparations, spike-recovery controls, calibration-verification standards, and quality-control samples are evaluated with each batch to confirm extraction efficiency, accuracy, precision, linearity, and absence of carryover or significant matrix interference.

Phospholipids are structurally diverse amphiphilic lipids that cannot be fully characterized by total phosphorus measurement or non-specific gravimetric lipid testing alone. LC-MS/MS provides molecular specificity that distinguishes phospholipid classes and individual acyl-chain species from triglycerides, sterols, glycolipids, free fatty acids, and formulation excipients. This distinction is especially important where a product claim relates to a particular phospholipid class, such as phosphatidylserine or phosphatidylcholine, or where the source and composition of a phospholipid-rich ingredient must be verified. The use of internal standards and MS/MS fragmentation improves quantitative reliability in complex matrices that may exhibit differential extraction recovery or ion-suppression effects. The assay supports incoming-material qualification, lot-to-lot consistency, label-claim substantiation, and dietary supplement cGMP quality-control requirements under 21 CFR 111.

This test quantifies piperine (1-[5-(1,3-benzodioxol-5-yl)-1-oxo-2,4-pentadienyl]piperidine) — the principal pungent alkaloid and primary bioactive constituent of black pepper (Piper nigrum L.) fruit — in raw materials, black pepper powders, extracts, and dietary supplements using High-Performance Liquid Chromatography (HPLC). Piperine is responsible for the characteristic pungency of black pepper and is widely recognized for its ability to enhance the bioavailability of co-administered nutrients and pharmaceuticals by inhibiting intestinal and hepatic metabolic enzymes. It is used both as a standalone active ingredient and as a bioavailability-enhancing adjunct in multi-ingredient supplement formulations. Accurate potency verification is essential for label claim substantiation and for confirming that the declared piperine content is present. Results are reported as a percentage or in milligrams per gram or per serving.

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

Piperine's conjugated diene chromophore provides strong and characteristic UV absorption at 343 nm, making HPLC with UV detection a highly sensitive and specific method for its quantification without the need for derivatization. Chromatographic separation on a C18 column resolves piperine from structurally related piperamide alkaloids — including its geometric isomer chavicine — and from matrix components in complex botanical extracts and finished supplement blends, ensuring that potency results reflect piperine content specifically. This method is applicable to both standardized black pepper extracts (including proprietary forms such as BioPerine®) and non-standardized black pepper ingredients, and supports label claim substantiation, raw material qualification, and cGMP compliance under 21 CFR 111.

This assay quantifies piperine, the active alkaloid in black pepper and the standardized extract marketed as BioPerine. Using HPLC, it verifies piperine content in capsules, blends, and bioenhanced formulations to ensure label accuracy and consistent dosing in absorption-support products.

Samples are extracted using alcohol-based solvents under controlled conditions and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified piperine 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 underdosing or ingredient substitution.

This assay quantifies total polysaccharides using UV-Visible spectrophotometry (UV-Vis). The method provides a rapid and reliable measure of total carbohydrate polymers present in natural extracts and raw materials.

Samples are hydrolyzed under controlled conditions, and the resulting solution is reacted with colorimetric reagents to produce a measurable chromophore. Absorbance is read at a specific wavelength using a UV-Vis spectrophotometer, and concentration is determined relative to a glucose or polysaccharide standard curve.

Results are reported as % w/w (raw materials) or mg/serving (finished products). Testing confirms standardized potency, supports label claims, and ensures consistency for polysaccharide-rich ingredients such as mushrooms, algae, and plant extracts.

This test confirms the identity of pomegranate fruit (Punica granatum L.) in raw materials, fruit powders, juice powders, peel extracts, and dietary supplements using High-Performance Thin-Layer Chromatography (HPTLC). Punica granatum, commonly known as pomegranate, is a widely used functional food and dietary supplement ingredient valued for its exceptional antioxidant capacity and rich content of bioactive polyphenols. Its characteristic phytochemical profile includes ellagitannins (punicalagins A and B, punicalin, pedunculagin), ellagic acid and ellagic acid glycosides, anthocyanins (delphinidin, cyanidin, and pelargonidin glycosides), flavonoids (quercetin, luteolin, kaempferol glycosides), and organic acids. HPTLC identity testing generates a characteristic chromatographic fingerprint — anchored by the distinctive ellagitannin and ellagic acid profile — that is compared against an authenticated P. granatum reference standard to confirm species identity and detect potential adulteration, substitution with other fruit materials, or blending with inferior pomegranate fractions.

A representative sample is accurately weighed and extracted using an appropriate solvent system (e.g., methanol or aqueous ethanol) to capture the characteristic polyphenol profile of P. granatum fruit, including ellagitannins, ellagic acid, and anthocyanins. The extract is applied alongside a certified pomegranate 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 ellagitannin, ellagic acid, and flavonoid constituents of P. granatum. After development, the plate is derivatized with an appropriate reagent (e.g., Natural Products Reagent A / NP/PEG for flavonoid and polyphenol visualization, or ferric chloride for tannin visualization) 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.

Pomegranate raw materials — particularly juice powders and peel extracts — are subject to adulteration and dilution with other fruit materials, added sugars, or inferior pomegranate fractions that may lack the characteristic ellagitannin profile responsible for the majority of pomegranate's documented antioxidant and health-promoting activity. HPTLC fingerprinting provides a holistic, multi-compound chromatographic identity confirmation — anchored by the distinctive punicalagin and ellagic acid band pattern characteristic of authentic P. granatum fruit — that is more discriminating than single-marker ellagic acid assays alone, enabling detection of substitution or adulteration that would not be apparent from potency testing of individual markers. This method aligns with USP botanical identity testing guidelines and supports cGMP compliance under 21 CFR 111, ensuring that only correctly identified raw materials are used in finished products.

This assay quantifies pregnenolone, a steroidal hormone precursor involved in the biosynthesis of progesterone, estrogens, and androgens. Using HPLC, it verifies pregnenolone content in raw materials and finished products to confirm label claims, ensure potency, and detect degradation or adulteration.

Samples are extracted using organic solvents and analyzed by HPLC with UV detection at a compound-specific wavelength. Quantification is performed using certified pregnenolone reference standards, with internal standard correction and duplicate injections to ensure accuracy and reproducibility.

Results are reported in mg per g or per serving. Values are compared with label claims and formulation targets to confirm potency and support consistent dosing across production batches.

This test quantifies total proanthocyanidins (condensed tannins) in botanical extracts, dietary supplements, and raw materials using UV-Vis spectrophotometry with a colorimetric derivatization method. Proanthocyanidins are oligomeric and polymeric flavanols found at high concentrations in grape seed, pine bark (Pycnogenol), cranberry, and other berry-derived extracts, and their total content is the primary potency and standardization marker for these ingredients. Results are reported as a percentage of proanthocyanidin content relative to extract weight, expressed as procyanidin equivalents, to support label claim verification and cGMP compliance.

A representative sample is weighed and extracted using a methanol or acetone-water solvent system with sonication. The extract is reacted with a chromogenic reagent — typically the DMAC (4-dimethylaminocinnamaldehyde) or vanillin-hydrochloric acid method — which produces a colored complex selectively with proanthocyanidins and flavan-3-ol units. Absorbance is measured by UV-Vis spectrophotometry at the appropriate wavelength (640 nm for DMAC; 500 nm for vanillin-HCl). Quantification is performed against a multi-point external calibration curve prepared from a certified procyanidin or catechin reference standard, and results are expressed as percentage proanthocyanidins relative to the initial sample weight.

Total proanthocyanidin content is the industry-standard potency marker for grape seed, pine bark, and berry-derived extracts, and its accurate measurement is essential for verifying standardization levels and substantiating label claims. UV-Vis spectrophotometry with colorimetric derivatization is the conventional and widely accepted method for total proanthocyanidin quantification, providing a practical and reproducible measure of extract potency for both raw material qualification and finished product release testing.

This assay quantifies proline-rich polypeptides (PRPs), immunomodulatory peptides found in colostrum and whey. Using LC-MS/MS, it detects bioactive PRPs to validate functional claims.

Peptides are extracted and analyzed by LC-MS/MS using mass-specific transitions for PRP sequences. Quantification uses peptide standards and internal calibration.

Results are reported in µg/g or µg per serving. Supports neuroimmune and adaptogenic product positioning.

This test measures the acid protease (SAP — Spectrophotometric Acid Protease) activity of protease enzyme preparations at pH 3.0 in raw materials and dietary supplements using the FCC (Food Chemicals Codex) standardized activity assay. Acid proteases are a class of proteolytic enzymes that function optimally under acidic conditions, making them particularly relevant for protein digestion support in the stomach environment. SAP activity is expressed in Spectrophotometric Acid Protease Units (SAP), where one SAP unit is defined as the amount of enzyme that liberates a defined quantity of tyrosine equivalents from a hemoglobin substrate per unit time under the specified assay conditions (pH 3.0, 40°C). Accurate activity verification is essential for confirming that enzyme ingredient potency meets label claim specifications and for ensuring consistent digestive support efficacy. Results are reported in SAP units per gram or per serving.

A representative sample is accurately weighed and dissolved in an appropriate cold buffer to prepare a working enzyme solution. The acid protease activity assay is performed according to the FCC monograph procedure: a defined volume of the enzyme solution is incubated with a hemoglobin substrate solution at pH 3.0 and 40°C for a precisely timed reaction period. The reaction is terminated by addition of trichloroacetic acid (TCA) to precipitate undigested protein, and the absorbance of the TCA-soluble hydrolysate — reflecting the concentration of liberated tyrosine and tyrosine-equivalent peptides — is measured spectrophotometrically at 275 nm. Enzyme activity is calculated from the absorbance reading using the FCC-defined unit calculation, referenced against a tyrosine standard curve. Reagent blanks and substrate controls are run concurrently to correct for non-enzymatic hydrolysis and background absorbance.

The FCC SAP assay is the standardized, industry-recognized method for acid protease activity measurement, providing a reproducible and internationally accepted unit of enzyme potency that enables direct comparison of activity across different enzyme sources, suppliers, and batches. Performing the assay at pH 3.0 specifically characterizes the gastric-phase proteolytic activity of the enzyme preparation, which is the most physiologically relevant activity parameter for acid protease ingredients used in digestive enzyme supplement formulations. FCC-defined activity units are the standard for enzyme ingredient labeling and quality specifications in the dietary supplement industry, supporting label claim substantiation and cGMP compliance under 21 CFR 111.

This assay determines total protein content in food samples by measuring nitrogen content and converting it to protein using a standard conversion factor. The Dumas method is a combustion-based technique valued for its speed, accuracy, and minimal chemical waste compared to traditional methods.

Samples are combusted in an oxygen-rich environment, and the released nitrogen is measured via thermal conductivity detection. The measured nitrogen is converted to protein content using a conversion factor (typically 6.25). Calibration with certified reference materials and duplicate analyses ensure reliability.

Results are reported as grams of protein per 100 g of food. These values are used for nutritional labeling and quality control. Consistent protein levels across batches confirm that the product meets formulation specifications, while deviations may indicate processing issues.

This test quantifies total protein content in food, dietary supplement, and raw material matrices using the Kjeldahl nitrogen determination method in accordance with AOAC Official Method 981.10 (Crude Protein in Meat and Meat Products, applicable broadly to food and supplement matrices). The Kjeldahl method measures total organic nitrogen in a sample, which is converted to protein content by multiplication with an appropriate nitrogen-to-protein conversion factor (commonly 6.25 for general food proteins, or matrix-specific factors such as 5.71 for wheat, 6.38 for dairy, and others as applicable). As a reference method with a long history of regulatory acceptance, Kjeldahl protein determination is recognized by AOAC International, the FDA, and international food regulatory bodies for nutritional labeling compliance under 21 CFR 101. Results are reported as a percentage of crude protein on an as-received or dry basis.

A representative sample is accurately weighed and subjected to acid digestion in concentrated sulfuric acid in the presence of a catalyst system (e.g., copper sulfate and potassium sulfate) at elevated temperature until complete oxidation of organic matter is achieved, converting all organic nitrogen to ammonium sulfate. The digested solution is made alkaline by addition of concentrated sodium hydroxide, and the liberated ammonia is distilled by steam distillation into a receiving solution of boric acid or a standard acid solution. The captured ammonia is quantified by back-titration with a standardized hydrochloric or sulfuric acid solution to a potentiometric or colorimetric endpoint. Total nitrogen content is calculated from the titration volume and expressed as a percentage; crude protein is calculated by multiplying the nitrogen percentage by the appropriate nitrogen-to-protein conversion factor. Reagent blanks are analyzed concurrently to correct for background nitrogen.

The Kjeldahl method is the long-established reference method for total protein determination in food and dietary supplement matrices, providing a direct measurement of total organic nitrogen that is independent of protein structure, molecular weight, or amino acid composition. AOAC 981.10 is a validated, internationally recognized official method with broad regulatory acceptance for nutritional labeling compliance. While the Kjeldahl method measures total nitrogen — including non-protein nitrogen from nucleic acids, free amino acids, and other nitrogenous compounds — it remains the standard for crude protein reporting in regulatory and labeling contexts. This method supports nutritional label compliance under 21 CFR 101, raw material specification testing, and cGMP compliance under 21 CFR 111.

This test determines the protein content of a sample on a dry weight basis by combining two complementary methods: total nitrogen quantification by the Dumas combustion method (nitrogen analyzer) and moisture determination by AOAC Official Method 925.10 (Loss on Drying). The Dumas method measures total nitrogen released upon high-temperature combustion of the sample, which is converted to protein content using an appropriate nitrogen-to-protein conversion factor. The moisture content determined by AOAC 925.10 is then used to correct the protein result to a dry basis, providing a moisture-independent measure of true protein content. This is the standard approach for protein specification and label claim verification in protein powders, amino acid ingredients, and high-protein food and supplement products. Results are reported as a percentage protein on a dry weight basis.

For moisture determination (AOAC 925.10), a representative sample is accurately weighed into a pre-dried, tared dish and dried in a convection oven at 105°C until constant weight is achieved. The moisture content is calculated from the loss in weight relative to the initial sample weight. For protein determination, a separate representative sample is accurately weighed and combusted at high temperature (typically above 900°C) in a nitrogen analyzer. The nitrogen gas released is measured by thermal conductivity detection, and total nitrogen content is calculated. Protein content is derived by multiplying total nitrogen by the appropriate conversion factor (e.g., 6.25 for general food proteins, or a product-specific factor where applicable). The protein result is then corrected to a dry basis using the moisture content determined by AOAC 925.10.

Reporting protein content on a dry basis eliminates the variability introduced by moisture differences between batches and between raw materials from different suppliers, providing a standardized and comparable measure of true protein content. The Dumas combustion method is the preferred modern alternative to the Kjeldahl method for total nitrogen determination, offering faster analysis, no hazardous reagents, and equivalent accuracy. Combining Dumas nitrogen analysis with AOAC 925.10 moisture correction is the industry-standard approach for protein specification in dietary supplement and food ingredient applications, supporting accurate nutrition labeling under 21 CFR 101 and raw material qualification under 21 CFR 111.

This assay quantifies protodioscin, a steroidal saponin commonly found in Tribulus terrestris and other botanical extracts. Using HPLC, it verifies protodioscin content in male health and performance supplements to ensure potency, authenticity, and consistent standardization.

Samples are extracted using methanol or aqueous alcohol 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 protodioscin standards, with internal standard correction and duplicate injections to ensure precision.

Results are reported in mg per g or per serving. Values are compared with label claims and formulation specifications to confirm standardized extract levels and detect adulteration or batch-to-batch variability.

Proximate analysis is a standardized suite of quantitative tests used to determine the macroscopic nutritional composition of food, beverage, and dietary supplement products. The standard panel includes the direct analytical determination of Moisture, Crude Protein, Crude Fat, and Ash, while Total Carbohydrates and Calories are determined by calculation. This comprehensive profile is the foundational requirement for generating Nutritional Facts or Supplement Facts panels and verifying macronutrient label claims. Moisture and Ash determine the water content and total inorganic mineral content, respectively. Protein is determined by measuring total nitrogen, and fat is measured via solvent extraction. Carbohydrates are calculated "by difference" (100% minus the sum of moisture, protein, fat, and ash). Calories are calculated using standardized Atwater factors (e.g., 4 kcal/g for protein and carbohydrates, 9 kcal/g for fat). This testing is critical for regulatory compliance under 21 CFR 101 (Food Labeling).

The panel utilizes a combination of standardized compendial methods (e.g., AOAC International). Moisture is typically determined by gravimetric loss on drying (LOD) in a vacuum oven or via Karl Fischer titration. Ash is determined by incinerating the sample in a muffle furnace (typically at 550–600°C) until all organic matter is destroyed and only inorganic residue remains, which is then weighed. Protein is quantified using the Kjeldahl method (e.g., AOAC 981.10) or Dumas combustion to measure total nitrogen, which is then multiplied by a product-specific conversion factor (commonly 6.25). Fat is measured via continuous solvent extraction (e.g., Soxhlet) or acid hydrolysis followed by extraction, depending on the complexity of the matrix. Carbohydrates are calculated by subtracting the measured percentages of moisture, ash, protein, and fat from 100%. Calories are calculated by multiplying the grams of protein, fat, and carbohydrates by their respective FDA-recognized Atwater caloric conversion factors. Quality control measures, including blank determinations and certified reference materials, are employed across all assays.

Proximate analysis is the universally accepted and regulatory-mandated methodology for nutritional profiling and labeling. Utilizing standardized compendial methods (AOAC) ensures that the results are robust, reproducible, and legally defensible. Calculating carbohydrates by difference and calories via Atwater factors is the standard FDA-approved approach under 21 CFR 101.9 for nutrition labeling. This complete panel provides manufacturers with the critical data required to formulate products accurately, verify macronutrient label claims, ensure lot-to-lot consistency, and meet both cGMP (21 CFR 111) and Nutrition Labeling and Education Act (NLEA) requirements.

This assay quantifies trans-pterostilbene, a methylated analog of resveratrol known for its enhanced bioavailability and antioxidant activity. Using HPLC, it verifies pterostilbene content in supplements and longevity-focused formulations to confirm label accuracy and dosing consistency.

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

Results are reported in mg per g or per serving. Values are compared to formulation targets and label claims to confirm potency and detect variability in raw material or storage stability.

This test quantifies Pyridoxal-5-Phosphate Monohydrate (P5P) — the biologically active, phosphorylated coenzyme form of Vitamin B6 — in dietary supplements and raw materials using UV-Visible (UV-Vis) Spectrophotometry. Unlike standard pyridoxine HCl, P5P is the metabolically active form that does not require hepatic conversion, making it a premium Vitamin B6 ingredient. Accurate potency verification is essential for confirming that the declared amount of this active coenzyme form is present and intact. Results are reported as a percentage or in milligrams per gram or per serving to support label claim substantiation and cGMP compliance.

A representative sample is accurately weighed and dissolved in a dilute aqueous buffer (typically phosphate buffer at a defined pH) to stabilize the phosphorylated compound and ensure complete dissolution. The absorbance of the solution is measured at the characteristic UV absorption maximum of P5P — approximately 388 nm in neutral to slightly alkaline conditions — against a solvent blank. Concentration is calculated using the Beer-Lambert law with the established molar extinction coefficient for P5P, or alternatively by comparison to a multi-point calibration curve prepared from a certified P5P reference standard. Measurements are performed in triplicate and the average value is used for final quantification.

Rationale | P5P is a high-value, premium form of Vitamin B6 that commands a price premium over standard pyridoxine HCl, making potency verification important for both label claim accuracy and raw material qualification. UV-Vis spectrophotometry is well suited for P5P quantification in high-purity raw material testing, where the compound is the dominant UV-absorbing species and matrix interference is minimal. The method is rapid, cost-effective, and appropriate for incoming material screening and release testing under 21 CFR 111. For complex finished product matrices where interfering compounds may be present, HPLC is the preferred alternative.

This assay quantifies pyrroloquinoline quinone (PQQ), a redox cofactor used in supplements targeting brain health, energy metabolism, and anti-aging. Using LC-MS/MS, it accurately measures PQQ content in capsules, powders, and functional formulations to verify label claims and ensure bioactive potency.

Samples are extracted using aqueous or alcohol-based solvents and analyzed by LC-MS/MS with compound-specific mass transitions. Quantification is performed using certified PQQ standards with internal standard correction and duplicate injections for precision and reproducibility.

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

This assay quantifies quercetin, a plant-derived flavonoid commonly used in immune, allergy, and longevity supplements. Using LC-MS/MS, it measures quercetin content in capsules, powders, and botanical blends to verify label claims and support bioactive consistency across formulations.

Samples are extracted in alcohol or methanol-based solvents and analyzed by LC-MS/MS using compound-specific mass transitions. Quantification is performed with certified quercetin standards, 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 dosing consistency and detect degradation or adulteration.

This assay measures raspberry ketones (4-(4-hydroxyphenyl)-2-butanone) using High-Performance Liquid Chromatography (HPLC). The method provides accurate and selective quantification of this aromatic compound in raw materials and finished formulations.

Samples are extracted and analyzed under validated HPLC chromatographic conditions. Raspberry ketones are separated from related phenolic compounds and detected via UV absorbance. Quantitation is performed using certified reference standards, with calibration curves and replicate injections ensuring accuracy and reproducibility.

Testing verifies label claims, confirms purity, and ensures consistent quality in raspberry-ketone–containing products.

This assay quantifies rauwolscine (also known as alpha-yohimbine), a stimulant alkaloid structurally related to yohimbine. Using HPLC, it verifies rauwolscine content in pre-workout, thermogenic, and focus-enhancing supplements to ensure accurate dosing and label compliance.

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 rauwolscine 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 with formulation targets and label claims to confirm potency and detect any over- or underdosing that could impact stimulant safety or regulatory compliance.