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.

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 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.

Method for the Determination of Lycopene in Supplements and Raw Material by Reversed-Phase Liquid Chromatography: Single-Laboratory Validation

2008 ◽  
Vol 91 (6) ◽  
pp. 1284-1297 ◽  
Author(s):  
André Müller ◽  
Bernd Pietsch ◽  
Nicole Faccin ◽  
Joseph Schierle ◽  
Edward H Waysek
Keyword(s):  
Dietary Supplements ◽  
High Performance ◽  
Raw Materials ◽  
Reversed Phase ◽  
Raw Material ◽  
Intermediate Precision ◽  
Relative Standard ◽  
Product Forms ◽  
Single Laboratory

Abstract A single-laboratory validation study was conducted for a liquid chromatographic (LC) method for the determination of total and all-trans-lycopene in a variety of dietary supplements and raw materials. Gelatin-based and other water-dispersible beadlets, or tablets, capsules, and softgels containing such product forms, were digested with protease. Alginate formulations and the respective applications were treated with an alkaline sodium EDTA acetate buffer to release lycopene from the matrix. Lycopene and other carotenoids were extracted from the resulting aqueous suspensions with dichloromethane and ethanol. Oily product forms were directly dissolved in dichloromethane and ethanol. The extracts were chromatographed on an isocratic high-performance LC system using a C16 alkylamide modified silica column that provided satisfactory resolution of all-trans-lycopene from its predominant cis-isomers and separated the lycopene isomers from other carotenoids such as - and -carotene, cryptoxanthin, lutein, and zeaxanthin. The within-day precision relative standard deviation (RSD) for the determination of total lycopene ranged from 0.9 to 5.7 over concentration ranges of 50200 g/kg for raw materials and 0.324 g/kg for dietary supplements. The intermediate precision RSD (total RSD) ranged from 0.8 to 8.9. Recoveries obtained for beadlet and tablet material for the different extraction variants ranged from 95.0 to 102.1 at levels of 0.0220 g/kg for tablets and from 95.0 to 101.1 at levels of 1200 g/kg for beadlet material.

scholarly journals Determination of Finasteride in Tablets by High Performance Liquid Chromatography

2007 ◽  
Vol 4 (1) ◽  
pp. 109-116 ◽  
Author(s):  
K. Basavaiah ◽  
B. C. Somashekar
Keyword(s):  
High Performance ◽  
Internal Standard ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Calibration Graph ◽  
Official Method ◽  
Pharmaceutical Dosage Forms ◽  
Bulk Drug ◽  
Liquid Chromatographic

A rapid, highly sensitive high performance liquid chromatographic method has been developed for the determination of finasteride(FNS) in bulk drug and in tablets. FNS was eluted from a ODS C18reversed phase column at laboratory temperature (30 ± 2°C) with a mobile phase consisting of methanol and water (80+20) at a flow rate of 1 mL min-1with UV detection at 225 nm. The retention time was ∼ 6.1 min and each analysis took not more than 10 min. Quantitation was achieved by measurement of peak area without using any internal standard. Calibration graph was linear from 2.0 to 30 μg mL-1with limits of detection (LOD) and quantification (LOQ) being 0.2 and 0.6 μg mL-1, respectively. The method was validated according to the current ICH guidelines. Within-day co efficients of variation (CV) ranged from 0.31 to 0.69% and between-day CV were in the range 1.2-3.2%. Recovery of FNS from the pharmaceutical dosage forms ranged from 97.89 – 102.9 with CV of 1.41-4.13%. The developed method was compared with the official method for FNS determination in its tablet forms.

Determination of tigecycline in turkey plasma by LC-MS/MS: validation and application in a pharmacokinetic study

2017 ◽  
Vol 20 (2) ◽  
pp. 241-249 ◽  
Author(s):  
A. Jasiecka-Mikołajczyk ◽  
J.J. Jaroszewski
Keyword(s):  
Pharmacokinetic Study ◽  
High Performance ◽  
Limit Of Detection ◽  
Internal Standard ◽  
Reversed Phase ◽  
Selected Reaction Monitoring ◽  
Limit Of Quantification ◽  
Complicated Skin ◽  
Intravenous And Oral Administration

Abstract Tigecycline (TIG), a novel glycylcycline antibiotic, plays an important role in the management of complicated skin and intra-abdominal infections. The available data lack any description of a method for determination of TIG in avian plasma. In our study, a selective, accurate and reversed-phase high performance liquid chromatography-tandem mass spectrometry method was developed for the determination of TIG in turkey plasma. Sample preparation was based on protein precipitation and liquid-liquid extraction using 1,2-dichloroethane. Chromatographic separation of TIG and minocycline (internal standard, IS) was achieved on an Atlantis T3 column (150 mm × 3.0 mm, 3.0 μm) using gradient elution. The selected reaction monitoring transitions were performed at 293.60 m/z → 257.10 m/z for TIG and 458.00 m/z → 441.20 m/z for IS. The developed method was validated in terms of specificity, selectivity, linearity, lowest limit of quantification, limit of detection, precision, accuracy, matrix effect, carry-over effect, extraction recovery and stability. All parameters of the method submitted to validation met the acceptance criteria. The assay was linear over the concentration range of 0.01-100 μg/ml. This validated method was successfully applied to a TIG pharmacokinetic study in turkey after intravenous and oral administration at a dose of 10 mg/kg at various time-points.

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