The analysis of thiamin and its derivatives in whole blood samples under high pH conditions of the mobile phase

Landeghem, Bart A.J. van; Puts, Johan; Claessens, H.A.

doi:10.1016/j.jchromb.2005.05.027

Cited by 13 publications

(12 citation statements)

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“…One group of 5 untreated animals was anesthetized and blood removed as vehicle control. Blood samples were directly processed to analyze benfotiamine, S-benzoylthiamine, thiamine and its derivatives by HPLC [66][67][68][69][70].…”

Section: Methodsmentioning

confidence: 99%

Efficient Transdermal Delivery of Benfotiamine in an Animal Model

2015

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We designed a transdermal system to serve as a delivery platform for benfotiamine utilizing the attributes of passive penetration enhancing molecules to penetrate through the outer layers of skin combined with the advance of incorporating various peripherally-acting vasodilators to enhance drug uptake. Benfotiamine, incorporated into this transdermal formulation, was applied to skin in an animal model in order to determine the ability to deliver this thiamine pro-drug effectively to the sub-epithelial layers. In this proof of concept study in guinea pigs, we found that a single topical application of either a solubilized form of benfotiamine (15 mg) or a microcrystalline suspension form (25 mg) resulted in considerable increases of the dephosphorylated benfotiamine (S-benzoylthiamine) in the skin tissue as well as in significant increases in the thiamine and thiamine phosphate pools compared to control animals. The presence of a ~8000x increase in thiamine and increases in its phosphorylated derivatives in the epidermis

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Section: Methodsmentioning

confidence: 99%

Efficient Transdermal Delivery of Benfotiamine in an Animal Model

2015

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“…Benfotiamine was measured in the whole blood with an assay modification described below based on the previously published methods [31][32][33][34]. The analytical system used for the quantitation of benfotiamine was an Agilent 1100 HPLC unit (Agilent, Santa Clara, CA) with fluorimetric detector connected to an Agilent ChemStation and data analysis unit.…”

Section: Methodsmentioning

confidence: 99%

Pharmacokinetics of the transdermal delivery of benfotiamine

Zhu

Váradi

Carter³

2015

Acta Diabetol

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“…The most frequently used method for analysis of TDP and PLP is a HPLC-based assay for each vitamin separately [ 11 – 25 ]. Indeed, the currently used HPLC-based methods in our laboratory for measuring TDP and PLP consist of two separate HPLC methods.…”

Section: Introductionmentioning

confidence: 99%

“…Indeed, the currently used HPLC-based methods in our laboratory for measuring TDP and PLP consist of two separate HPLC methods. TDP is analyzed at pH 10 by reversed phase liquid chromatography performing a pre-column derivatization step with alkaline potassium ferricyanide and fluorescence detection as described by van Landeghem et al [ 11 ]. PLP is pre-column converted to a semi-carbazone followed by column separation and fluorescence detection at pH12 according to Schrijver et al [ 12 ].…”

Section: Introductionmentioning

confidence: 99%

Simultaneous Determination of Underivatized Vitamin B1 and B6 in Whole Blood by Reversed Phase Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry

et al. 2015

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BackgroundVitamin B1 (thiamine-diphosphate) and B6 (pyridoxal-5’phosphate) are micronutrients. Analysis of these micronutrients is important to diagnose potential deficiency which often occurs in elderly people due to malnutrition, in severe alcoholism and in gastrointestinal compromise due to bypass surgery or disease. Existing High Performance Liquid Chromatography (HPLC) based methods include the need for derivatization and long analysis time. We developed an Ultra High Performance Liquid Chromatography Tandem Mass spectrometry (UHPLC-MS/MS) assay with internal standards for simultaneous measurement of underivatized thiamine-diphosphate and pyridoxal-5’phosphate without use of ion pairing reagent.MethodsWhole blood, deproteinized with perchloric acid, containing deuterium labelled internal standards thiamine-diphosphate(thiazole-methyl-D3) and pyridoxal-5’phosphate(methyl-D3), was analyzed by UHPLC-MS/MS. The method was validated for imprecision, linearity, recovery and limit of quantification. Alternate (quantitative) method comparisons of the new versus currently used routine HPLC methods were established with Deming regression.ResultsThiamine-diphosphate and pyridoxal-5’phosphate were measured within 2.5 minutes instrumental run time. Limits of detection were 2.8 nmol/L and 7.8 nmol/L for thiamine-diphosphate and pyridoxal-5’phosphate respectively. Limit of quantification was 9.4 nmol/L for thiamine-diphosphate and 25.9 nmol/L for pyridoxal-5’phosphate. The total imprecision ranged from 3.5–7.7% for thiamine-diphosphate (44–157 nmol/L) and 6.0–10.4% for pyridoxal-5’phosphate (30–130 nmol/L). Extraction recoveries were 101–102% ± 2.5% (thiamine-diphosphate) and 98–100% ± 5% (pyridoxal-5’phosphate). Deming regression yielded slopes of 0.926 and 0.990 in patient samples (n = 282) and national proficiency testing samples (n = 12) respectively, intercepts of +3.5 and +3 for thiamine-diphosphate (n = 282 and n = 12) and slopes of 1.04 and 0.84, intercepts of -2.9 and +20 for pyridoxal-5’phosphate (n = 376 and n = 12).ConclusionThe described UHPLC-MS/MS method allows simultaneous determination of underivatized thiamine-diphosphate and pyridoxal-5’phosphate in whole blood without intensive sample preparation.

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The analysis of thiamin and its derivatives in whole blood samples under high pH conditions of the mobile phase

Cited by 13 publications

References 20 publications

Efficient Transdermal Delivery of Benfotiamine in an Animal Model

Efficient Transdermal Delivery of Benfotiamine in an Animal Model

Pharmacokinetics of the transdermal delivery of benfotiamine

Simultaneous Determination of Underivatized Vitamin B1 and B6 in Whole Blood by Reversed Phase Ultra High Performance Liquid Chromatography Tandem Mass Spectrometry

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