کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10551015 | 967104 | 2005 | 8 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Quantification of sugar phosphate intermediates of the pentose phosphate pathway by LC-MS/MS: application to two new inherited defects of metabolism
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کلمات کلیدی
FBSd-Ribulose-5-phosphated-glucose-6-phosphateACNd-fructose-6-phosphaterPIPPPDHAPMRMHBSSLC–MS/MS - LC-MS / MSP/S - P / Sinternal standard - استاندارد داخلیAcetonitrile - استونیتریلTransaldolase - ترانسالدولازdihydroxyacetone phosphate - دی هیدروکسی استون فسفاتRibose-5-phosphate isomerase - ریبوز 5-فسفات ایزومرازfetal bovine serum - سرم جنین گاوSugar phosphate - شکر فسفاتHanks’ balanced salt solution - محلول نمک متعادل هانکسpentose phosphate pathway - مسیر پنتوز فسفاتmultiple reaction monitoring - نظارت چندگانه چندگانهpenicillin and streptomycin - پنی سیلین و استرپتومایسینLiquid chromatography with tandem mass spectrometry - کروماتوگرافی مایع با طیف سنجی جرمی دو طرفه
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
We describe a liquid chromatography tandem mass spectrometry (LC-MS/MS) method to quantify pentose phosphate pathway intermediates (triose-3-phosphates, tetrose-4-phosphate, pentose-5-phosphate, pentulose-5-phosphates, hexose-6-phosphates and sedoheptulose-7-phosphate (sed-7P)) in bloodspots, fibroblasts and lymphoblasts. Liquid chromatography was performed using an ion pair loaded C18 HPLC column and detection of the sugar phosphates was carried out by tandem mass spectrometry using an electron ion spray source operating in the negative mode and multiple reaction monitoring. Reference values for the pentose phosphate pathway intermediates in blood spots, fibroblasts and lymphoblasts were established. The method was applied to cells from patients affected with a deficiency of transaldolase. The transaldolase-deficient cells showed an increased concentration of sedoheptulose-7-phosphate. (Bloodspots: 5.19 and 5.43 μmol/L [0.49-3.33 μmol/L]; fibroblasts 7.43 and 26.46 μmol/mg protein [0.31-1.14 μmol/mg protein]; lymphoblasts 16.03 μmol/mg protein [0.61-2.09 μmol/mg protein].) The method was also applied to study enzymes of the pentose phosphate pathway by incubating fibroblasts or lymphoblasts homogenates with ribose-5-phosphate or 6-phosphogluconate and the subsequent analysis of the formed sugar phosphates.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography B - Volume 823, Issue 1, 25 August 2005, Pages 18-25
Journal: Journal of Chromatography B - Volume 823, Issue 1, 25 August 2005, Pages 18-25
نویسندگان
Mirjam M.C. Wamelink, Eduard A. Struys, Jojanneke H.J. Huck, Birthe Roos, Marjo S. van der Knaap, Cornelis Jakobs, Nanda M. Verhoeven,