کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1212979 1494107 2013 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Quantification of urinary folate catabolites using liquid chromatography–tandem mass spectrometry
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Quantification of urinary folate catabolites using liquid chromatography–tandem mass spectrometry
چکیده انگلیسی


• Urinary folate catabolites are useful as non-invasive markers of folate status.
• Low concentrations and interfering compounds complicate quantification.
• Derivatization to butyl esters improves ionization efficiency and separation.
• UPLC–APCI-MS/MS method provides high accuracy, sensitivity and specificity.
• The assay is extensively validated and optimized for cost-effective high-throughput.

Folate catabolites p-aminobenzoylglutamate (pABG) and p-acetamidobenzoylglutamate (apABG) in human urine result from break-down of endogenous folate pools and are potential biomarkers of folate status. There is growing interest in analysis of these non-invasive indicators of folate status, since widespread diseases such as cancer, arteriosclerosis and dementia may be linked to disturbed availability of folates. Determination of pABG and apABG in human urine is challenging due to their low urinary concentrations and due to interferences with other urinary compounds. To address these analytical difficulties, we developed an improved LC–MS/MS method with chemical derivatization for fast, selective and sensitive quantification of pABG and apABG in human urine. Forming butyl esters of urinary folate catabolites pABG and apABG improves ionization efficiency as well as enables selective chromatographic separation on standard C18 reversed-phase column material. In contrast to some previously proposed methods for folate catabolites, the new method allows precise differentiation of apABG from pABG. Partial degradation of apABG during derivatization is exactly accounted for using a second differentially labeled stable isotope internal standard. This method is highly sensitive and covers the full range of physiologically occurring concentrations (from 2 to 1000 nmol/L), with volume requirements of only 80 μL urine. Method performance has been validated according to widely accepted standard recommendations. Use of two stable isotope-labeled internal standards and qualifier ion monitoring for both analytes ensure correct identification and unbiased quantification. With run times of less than 2.5 min per sample and cost-efficient sample preparation, this method allows exact quantitation of urinary folate catabolites pABG and apABG for large-scale non-invasive screening of folate status in clinical and epidemiological trials.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 929, 15 June 2013, Pages 116–124
نویسندگان
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