A single-step method for simultaneous quantification of thiamine and its phosphate esters in whole blood sample by ultra-performance liquid chromatography-mass spectrometry

Feasibility of Mass-Spectrometry to Lower Cost and Blood Volume Requirements for Assessment of B Vitamins in Patients Undergoing Bariatric Surgery

“…Because of our research on the impact of bariatric surgery on anemia induced by iron and copper deficiencies [21,22], we sought to measure B vitamins as part of a nutritional panel, since deficiencies in many B vitamins lead to this complication. The literature contains recent research on HPLC-MS/MS technologies for simultaneous assessment of B vitamins from human biological fluids including serum/plasma [9][10][11], whole blood [6][7][8], blood spots [23], and breast milk [24]. The analysis in each of these cases required less than 500 µL of blood, which is less than the 1000 µL required for HPLC techniques that did not use mass spectrometry [3].…”

“…Another matrix issue we may consider when analyzing B vitamins in the future is the suitability of plasma versus whole blood. For example, while Redeuil et al successfully measured thiamine and its metabolites in plasma [9], other groups preferred using whole blood [6][7][8], since the concentration of thiamine diphosphate in erythrocytes is a better measure of thiamine status. Finally, since we expect to use the HPLC-MS/MS method to comprehensively assess the status of B vitamins in patients undergoing bariatric surgery, we would include assessment of vitamin B 12 in future analysis.…”

“…An exciting development is to couple HPLC with tandem mass spectrometry (MS/MS) for detection of the vitamin analytes, which is preferable to ultraviolet-visible spectroscopy or photodiode array detectors in terms of sensitivity and specificity [5]. Several research groups have successfully simultaneously measured B vitamins in blood using HPLC-MS/MS [6][7][8][9]. Some groups went further to apply the HPLC-MS/MS methodology to simultaneously quantify B vitamins in patient populations including those with colorectal cancer [10] and neural tube defects [11].…”

Utility of Whole Blood Thiamine Pyrophosphate Evaluation in TPK1-Related Diseases

“…Regardless of the methodology, the most common derivative measured in blood is TPP given that this is the most abundant form of Vitamin B1 present. However, additional phosphorylated and unphosphorylated forms of thiamine are observed using HPLC chromatograms and can be identified with reference to standards, albeit at low abundance, and sometimes at the limit of quantitation [13,16]. Recently described methods [16,17,18], using HPLC with fluorescence or mass spectrometric detection, can quantify other forms of thiamine.…”

“…However, additional phosphorylated and unphosphorylated forms of thiamine are observed using HPLC chromatograms and can be identified with reference to standards, albeit at low abundance, and sometimes at the limit of quantitation [13,16]. Recently described methods [16,17,18], using HPLC with fluorescence or mass spectrometric detection, can quantify other forms of thiamine. These methods have potential utility during the investigation of inborn errors of thiamine metabolism using biological samples where other thiamine species are more abundant.…”