Single-Laboratory Validation of a Method for the Determination of Vitamin D3 in Dietary Supplements by Liquid Chromatography with Tandem Mass Spectrometry (LC-MS/MS)

2014 ◽  
Vol 97 (2) ◽  
pp. 403-408
Author(s):  
Victor K M Lam ◽  
Ray C T Hung ◽  
Ella L M Wong ◽  
Johnny Y W Fok ◽  
Yiu-Chung Wong
Keyword(s):  
Formic Acid ◽  
Dietary Supplements ◽  
Vitamin D3 ◽  
Time Window ◽  
Collaborative Study ◽  
Internal Standard ◽  
Ammonium Formate ◽  
Soft Gels ◽  
Single Laboratory

Abstract A single-laboratory validation (SLV) for the analysis of vitamin D3 was performed in four types of dietary supplements (capsules, soft gels, syrups, and tablets) using LC-MS/MS. Samples were treated by alkaline saponification for oil-based soft gels and utilized EDTA solution for capsules, syrups,and tablets prior to n-hexane extraction. Vitamin D3 in sample extracts was separated on a reversed-phase C18 column (100 × 2.1 mm, 2.7 μm) using a mobile phase of a 95 + 5 (v/v) mixture of 5 mM ammonium formate in methanol containing 0.1% formic acid and 5 mM ammonium formate in 0.1% formic acid running at a flow rate of 0.2 mL/min. Vitamin D3 was confirmed by the presence of three fragment ions at m/z 107, 159, and 259 within a defined retention time window from the precursor ion at m/z 385. Quantitation was based on the peak area at m/z 367 to that of the internal standard (d3-vitamin D3) at m/z 370 with reference to the respective response ratios of the calibration standards. The linear response of vitamin D3 ranged from 0.10 to 6.29 mg/L and the correlation coefficient (r) of the six-point calibrationcurves was >0.999. Accuracy, in terms of the spiked recoveries from blank syrup and starch powder at three different concentration levels, was 101–103%. Precision, determined by two different analysts over a period of 5 weeks, ranged from 2.7 to 7.0%for the four preparations. The SLV demonstrates the present LC-MS/MS method is reliable and robust for the determination of vitamin D3 in the studied dietary supplements. Considering the attainmentof satisfactory SLV results, further validation through intra-laboratory collaborative study is recommended.

scholarly journals Single-Laboratory Validation of a High-Performance Liquid Chromatographic-Diode Array Detector-Fluorescence Detector/Mass Spectrometric Method for Simultaneous Determination of Water-Soluble Vitamins in Multivitamin Dietary Tablets

2009 ◽  
Vol 92 (2) ◽  
pp. 680-688 ◽  
Author(s):  
Pei Chen ◽  
Renata Atkinson ◽  
Wayne R Wolf
Keyword(s):  
Formic Acid ◽  
Dietary Supplements ◽  
High Performance ◽  
Array Detector ◽  
Diode Array ◽  
Diode Array Detector ◽  
Fluorescence Detector ◽  
Liquid Chromatographic ◽  
Single Laboratory

Abstract The purpose of this study was to develop a single-laboratory validated (SLV) method using high-performance liquid chromatography with different detectors diode array detector (DAD); fluorescence detector (FLD); and mass spectrometry (MS) for determination of 7 B-complex vitamins (B1-thiamin, B2-riboflavin, B3-nicotinamide, B6-pyridoxine, B9-folic acid, pantothenic acid, and biotin) and vitamin C in multivitamin/multimineral dietary supplements. The method involves the use of a reversed-phase octadecylsilyl column (4 m, 250 2.0 mm id) and a gradient mobile phase profile. Gradient elution was performed at a flow rate of 0.25 mL/min. After a 5 min isocratic elution at 100 A (0.1 formic acid in water), a linear gradient to 50 A and 50 B (0.1 formic acid in acetonitrile) at 15 min was employed. Detection was performed with a DAD as well as either an FLD or a triple-quadrupole MS detector in the multiple reaction monitoring mode. SLV was performed using Standard Reference Material (SRM) 3280 Multivitamin/Multimineral Tablets, being developed by the National Institute of Standards and Technology, with support by the Office of Dietary Supplements of the National Institutes of Health. Phosphate buffer (10 mM, pH 2.0) extracts of the NIST SRM 3280 were analyzed by the liquid chromatographic (LC)-DAD-FLD/MS method. Following extraction, the method does not require any sample cleanup/preconcentration steps except centrifugation and filtration.

Expansion of the Scope of AOAC First Action Method 2012.25—Single-Laboratory Validation of Triphenylmethane Dye and Leuco Metabolite Analysis in Shrimp, Tilapia, Catfish, and Salmon by LC-MS/MS

2015 ◽  
Vol 98 (3) ◽  
pp. 636-648 ◽  
Author(s):  
Wendy C Andersen ◽  
Christine R Casey ◽  
Marilyn J Schneider ◽  
Sherri B Turnipseed
Keyword(s):  
Matrix Effects ◽  
Collaborative Study ◽  
Internal Standard ◽  
Triphenylmethane Dyes ◽  
Calibration Methods ◽  
Leucocrystal Violet ◽  
Leucomalachite Green ◽  
Method Accuracy ◽  
Single Laboratory

Abstract Prior to conducting a collaborative study of AOAC First Action 2012.25 LC-MS/MS analytical method for the determination of residues of three triphenylmethane dyes (malachite green, crystal violet, and brilliant green) and their metabolites (leucomalachite green and leucocrystal violet) in seafood, a single-laboratory validation of method 2012.25 was performed to expand the scope of the method to other seafood matrixes including salmon, catfish, tilapia, and shrimp. The validation included the analysis of fortified and incurred residues over multiple weeks to assess analyte stability in matrix at –80°C, a comparison of calibration methods over the range 0.25 to 4 μg/kg, study of matrix effects for analyte quantification, and qualitative identification of targeted analytes. Method accuracy ranged from 88 to 112% with 13% RSD or less for samples fortified at 0.5, 1.0, and 2.0 μg/kg. Analyte identification and determination limits were determined by procedures recommended both by the U. S. Food and Drug Administration and the European Commission. Method detection limits and decision limits ranged from 0.05 to 0.24 μg/kg and 0.08 to 0.54 μg/kg, respectively. AOAC First Action Method 2012.25 with an extracted matrix calibration curve and internal standard correction is suitable for the determination of triphenylmethane dyes and leuco metabolites in salmon, catfish, tilapia, and shrimp by LC-MS/MS at a residue determination level of 0.5 μg/kg or below.

Determination of Pyridoxine in Dietary Supplements by Liquid Chromatography with UV, Fluorescence, and Mass Spectrometric Detection: Single-Laboratory Validation

2013 ◽  
Vol 96 (2) ◽  
pp. 265-275 ◽  
Author(s):  
Robert J Goldschmidt ◽  
Wayne R Wolf
Keyword(s):  
Dietary Supplements ◽  
Vitamin B6 ◽  
Internal Standard ◽  
Reversed Phase ◽  
Reversed Phase Chromatography ◽  
Uv Fluorescence ◽  
Ms Detection ◽  
Different Types ◽  
Single Laboratory

Abstract A single-laboratory validation was performed for a method that determines pyridoxine, one of the B6 vitamers, in dietary supplements using LC and UV, fluorescence, or MS detection. The method was adapted for use with either HPLC or ultra-performance LC (UPLC). Pyridoxine is extracted from samples using 0.1 M formic acid, and specific conditions are adjusted for each of the different types of supplement materials examined. Reversed-phase chromatography with C18-based columns is used in both HPLC and UPLC. Fluorescence detection, often used in chromatographic analyses of vitamin B6 in foods, was successfully used here, but offered no great advantages over UV detection in the supplement materials tested. MS detection was also satisfactory, although use of an internal standard was required. Accuracy of the method was demonstrated in several ways, including use of a standard reference material. Precision and repeatability of the method were found acceptable by analysis of variance and HorRat repeatability calculations.

scholarly journals Method for the Determination of β-Carotene in Supplements and Raw Materials by Reversed-Phase Liquid Chromatography: Single Laboratory Validation

2004 ◽  
Vol 87 (5) ◽  
pp. 1070-1082 ◽  
Author(s):  
Joseph Schierle ◽  
Bernd Pietsch ◽  
Alan Ceresa ◽  
Christian Fizet ◽  
Edward H Waysek
Keyword(s):  
Liquid Chromatography ◽  
Dietary Supplements ◽  
Raw Materials ◽  
Collaborative Study ◽  
Reversed Phase ◽  
Raw Material ◽  
Relative Standard ◽  
Single Laboratory ◽  
Β Carotene

Abstract A single laboratory validation (SLV) study was conducted for a liquid chromatography (LC) method for the determination of total and all-trans-β-carotene in a variety of dietary supplements, including multivitamin tablets, softgels, capsules, and beadlet raw materials. Extraction variants were developed for the different types of supplements tested based upon the supplement type and level of β-carotene. Water dispersible formulations such as powders, emulsions, tablets, and capsules were enzymatically digested with protease and extracted with dichloromethane–ethanol. Oily suspensions were directly dissolved in dichloromethane–ethanol. After appropriate dilution or concentration, the extracts were chromatographed by using either a reversed-phase C18 column or, in products containing high amounts of α-carotene, a reversed-phase C30 column. The LC systems provided linear responses in the range of 0.1–50 μg β-carotene/mL. The main geometrical isomers of β-carotene (all-trans, 9-cis, 13-cis, and 15-cis) were well separated from each other and from other carotenoids such as α-carotene, cryptoxanthin, lutein, lycopene, and zeaxanthin. Duplicate determinations of total β-carotene performed by 2 technicians in 8 different test materials on 5 different days resulted in relative standard deviations of 1.2–4.4%. Recoveries determined for supplements and beadlet raw material spiked with β-carotene levels of 10 μg to 100 mg/test portion and 0.2–40%, respectively, ranged from 97.5 to 102.1%. On the basis of the accuracy, precision, and recovery results from the SLV study, the method is suggested for a collaborative study on the determination of total and all-trans-β-carotene in dietary supplements.

scholarly journals Determination of Ephedrine Alkaloids in Dietary Supplements and Botanicals by Liquid Chromatography/Tandem Mass Spectrometry: Collaborative Study

2003 ◽  
Vol 86 (4) ◽  
pp. 657-668 ◽  
Author(s):  
William A Trujillo ◽  
Wendy R Sorenson ◽  
J Laurensen ◽  
G Luo ◽  
R McClanahan ◽  
...  
Keyword(s):  
Liquid Chromatography ◽  
Dietary Supplements ◽  
Collaborative Study ◽  
Internal Standard ◽  
Negative Control ◽  
Interlaboratory Study ◽  
Tandem Mass ◽  
Accuracy And Precision ◽  
Ephedra Nevadensis

Abstract An interlaboratory study was conducted to evaluate the accuracy and precision of a method for ephedrine-type alkaloids [i.e., norephedrine (NE), norpseudoephedrine (NPE), ephedrine (E), pseudoephedrine (PE), methylephedrine (ME), and methylpseudoephedrine (MPE)] in dietary supplements and botanicals. The amount of ephedrine-type alkaloids present was determined using liquid chromatography with tandem mass selective detection. The samples were diluted to reflect a concentration of 0.0200 to 1.00 μg/mL for each alkaloid. An internal standard was added and the alkaloids were separated using a 5 μm phenyl LC column with an ammonium acetate, glacial acetic acid, acetonitrile, and water mobile phase. Eight blind duplicates of dietary supplements or botanicals were analyzed by 10 collaborators. Included was a negative control, ephedra nevadensis, and negative controls fortified at 2 different levels with each of the 6 ephedrine-type alkaloids. The spike levels were approximately 100 and 1000 μg/g for NE, 100 and 600 μg/g for NPE, 6500 and 65 000 μg/g for E, 1000 and 10 000 μg/g for PE, 300 and 3000 μg/g for ME, and 100 and 1000 μg/g for MPE. On the basis of the accuracy and precision results for this interlaboratory study, it is recommended that this method be adopted Official First Action for the determination of 6 different individual ephedrine-type alkaloids in dietary supplements and botanicals.

Determination of Total Choline by Liquid Chromatography– Electrospray Ionization–Tandem Mass Spectrometry in Infant Formulas

2012 ◽  
Vol 95 (1) ◽  
pp. 157-162 ◽  
Author(s):  
Shishan Fu ◽  
Baohua Tao ◽  
Shiyun Lai ◽  
Jingshun Zhang ◽  
Ren Yiping
Keyword(s):  
Electrospray Ionization ◽  
Neural Development ◽  
Internal Standard ◽  
Water Soluble ◽  
Infant Formulas ◽  
Research Areas ◽  
Hydrolyzed Protein ◽  
Evaluation Matrix ◽  
Single Laboratory

Abstract Choline is a water-soluble nutrient important for infants' brain and neural development. In infant formulas, choline is one of the important fortified nutrients. A single-laboratory validation study conducted an LC-electrospray ionization-MS/MS to determine total choline in infant formulas. Sample preparation was adopted from AOAC Official MethodSM 999.14, and instrumental running conditions were optimized. The LOQ was 0.2 μg/100 g, which is significant for measuring total choline in infant formulas. Average recoveries for milk-, rice-, soybean-, and hydrolyzed protein-based samples ranged from 86.45 ± 6.04% to 108.98 ± 3.68%, with RSD less than 7%. The repeatability RSD (RSDr) range was 0.24–3.59% in within-day evaluation and 1.16–3.24% in day-to-day evaluation. Matrix effect was also investigated, and can be effectively eliminated by using an internal standard. Therefore, this method has high credibility, and could be used as a routine method of quality control, or for clinical studies and other research areas.

Liquid Chromatographic Determination of Diazepam in Tablets: Collaborative Study

1985 ◽  
Vol 68 (3) ◽  
pp. 545-546
Author(s):  
Michael Tsougros
Keyword(s):  
Collaborative Study ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
Reverse Phase ◽  
Photometric Detection ◽  
Liquid Chromatographic Method ◽  
Liquid Chromatographic Determination ◽  
The Mean ◽  
Liquid Chromatographic

Abstract A stability indicating liquid chromatographic method for the determination of diazepam in tablets was collaboratively studied by 6 laboratories. The method uses a Cig reverse phase column, a methanolwater mobile phase, p-tolualdehyde as the internal standard, and photometric detection at 254 nm. The collaborators were supplied with a synthetic tablet powder and 3 commercial tablet samples. The mean recovery of diazepam from the synthetic tablet powder was 100.2%. For all samples analyzed, the coefficient of variation was < 1.5%. The method has been adopted official first action.

Determination of total proteinogenic amino acids and taurine by pre-column derivatisation and UHPLC: Single Laboratory Validation, First Action 2019.09

2020 ◽  
Author(s):  
George Joseph ◽  
Asha Varughese ◽  
Ann Daniel
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Infant Formula ◽  
Retention Time ◽  
Sulfonic Acid ◽  
Internal Standard ◽  
The Other ◽  
Method Performance ◽  
Single Laboratory

Abstract Background A method has been developed and validated for selective, accurate and precise determination of total proteinogenic amino acids and taurine from Infant Formula and Adult/Pediatric Nutritional Formulas (powders, ready-to-feed liquids, and liquid concentrates). The method was reviewed by the AOAC INTERNATIONAL SPIFAN Expert Review Panel (ERP) during the 133rd AOAC Annual Meeting & Expo on September 7, 2019 in Denver, CO, USA and was recommended to First Action Official MethodsSM status. Objective The method involves protein hydrolysis to amino acids, a simple pre-column derivatisation of amino acids and separation of derivatised amino acids by UHPLC. The quantification of amino acids is performed by multi-point calibration using norvaline as the internal standard. The analytical method is capable of quantitative determination for 22 proteinogenic amino acids, but cannot be used to quantitate tryptophan, which is destroyed during the acid hydrolysis step. Asparagine is determined as aspartic acid and glutamine as glutamic acid. The cystine and cysteine are converted to S-2-carboxyethylthiocysteine (CYSx) and the derivative is separated from the other amino acids. Citrulline which is present in some matrices and it is separated from other amino acids is not included in the method performance evaluation in the single laboratory validation (SLV). Method The proposed method met all the performance requirement limits set in standard method performance requirements (SMPR) 2014.013 for total proteinogenic amino acids and taurine. The correlation coefficient of multi-point calibration was not less than 0.999 for any amino acids at any point in the SLV study confirming the validity of linear dynanic range (LDR) and linearity of the method. The individual amino acids in the chromatogram were identified by absolute retention time and relative retention time (RRT) with respect to the internal standard norvaline. There were no significant (S/N Ratio <10) interferences from the reagents or by-products of derivatisation and targeted matrices. The method demonstrated high selectivity. Result Accuracy of the method was validated using standard reference materials (NIST SRM 1869 and 1849a) and spike recovery studies. The amino acid results in the SRMs were within the ranges of Reference Mass Fraction Values. The accuracy of the method was corroboratively validated by spike recovery studies. The average spike recovery range between 93 to 107% ensure the accuracy of the method for amino acids and compliance to the AOAC SMPR 2014.013. Conclusions Precision data of the method demonstrate that it meets the stakeholder requirements as per the SMPR. The mean RSDr for all the amino acids for 17 matrices selected for the SLV were not more than 4%. The method is very sensitive and the LOQ can go down to approximately ten times lower than the SMPR requirements. The sensitivity of method is a direct reflection of its signal to noise ratio which ensures guaranteed method performance at the lower levels of amino acids in these matrices. Highlights Taurine (aminoethane sulfonic acid) unlike the other amino acids is a beta-sulfonic amino acid that is not used in protein synthesis but is found as a free amino acid in tissues. The acidic functional group (-COOH) in common amino acid is replaced with a sulfonic acid (-SO3H) group in Taurine. The method offers baseline separation of citrulline which is an alpha amino acid generally present in Infant Formula and Adult/Pediatric Nutritional products. The separation of citrulline eliminates the risk of interference of this compound with other amino acids. The method can also separate and quantitate hydroxyproline, an important component of collagen that is often used to quantitate collagen. The method is simple and does not include any proprietary chemicals or instruments and can be performed on any basic reverse phase UHPLC system with UV detection.

Liquid Chromatographic Determination of Methiocarb in Technical and Formulated Products: Collaborative Study

1984 ◽  
Vol 67 (3) ◽  
pp. 492-493
Author(s):  
Stephen C Slahck ◽  
◽  
A A Carlstrom ◽  
L T Chenery ◽  
N D Ellis ◽  
...  
Keyword(s):  
Coefficient Of Variation ◽  
Collaborative Study ◽  
Internal Standard ◽  
Chromatographic Determination ◽  
Reverse Phase ◽  
Reverse Phase Chromatography ◽  
Wettable Powder ◽  
Liquid Chromatographic Determination ◽  
Liquid Chromatographic

Abstract An LC method for the determination of methiocarb in methiocarb technical and formulated products has been subjected to a collaborative study with 9 participating collaborators. Formulations are extracted with acetonitrile and analyzed by reverse phase chromatography, with acetophenone as an internal standard. Collaborators were furnished samples of technical, 75% wettable powder, 75% seed treater, 75% concentrate, and 50% hopper box treater. Coefficient of variation values obtained on the 5 samples were 0.71, 0.83, 0.62, 1.57, and 0.82%, respectively. The method has been adopted official first action.

Sign in / Sign up

Export Citation Format

Share Document