کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1251547 1496283 2016 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A facile stable-isotope dilution method for determination of sphingosine phosphate lyase activity
ترجمه فارسی عنوان
یک روش رقیق کننده پایدار با ایزوتوپ برای تعیین فعالیت اسپینگزین فسفات لیاز
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی (عمومی)
چکیده انگلیسی

A new technique for quantifying sphingosine phosphate lyase activity in biological samples is described. In this procedure, 2-hydrazinoquinoline is used to convert (2E)-hexadecenal into the corresponding hydrazone derivative to improve ionization efficiency and selectivity of detection. Combined utilization of liquid chromatographic separation and multiple reaction monitoring-mass spectrometry allows for simultaneous quantification of the substrate S1P and product (2E)-hexadecenal. Incorporation of (2E)- d5-hexadecenal as an internal standard improves detection accuracy and precision. A simple one-step derivatization procedure eliminates the need for further extractions. Limits of quantification for (2E)-hexadecenal and sphingosine-1-phosphate are 100 and 50 fmol, respectively. The assay displays a wide dynamic detection range useful for detection of low basal sphingosine phosphate lyase activity in wild type cells, SPL-overexpressing cell lines, and wild type mouse tissues. Compared to current methods, the capacity for simultaneous detection of sphingosine-1-phosphate and (2E)-hexadecenal greatly improves the accuracy of results and shows excellent sensitivity and specificity for sphingosine phosphate lyase activity detection.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Chemistry and Physics of Lipids - Volume 194, January 2016, Pages 101–109
نویسندگان
, , , ,