کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10533712 | 961890 | 2012 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
High-performance liquid chromatography-tandem mass spectrometry assay of fatty acid amide hydrolase (FAAH) in blood: FAAH inhibition as clinical biomarker
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کلمات کلیدی
BioanalyticalArachidonoylethanolamideHPLC–MS/MS - HPLC-MS / MSEnzyme activity assay - آزمایش فعالیت آنزیمیanandamide - آناندامیدfatty acid amide hydrolase (FAAH) - اسید آمید هیدرولاز اسید چرب (FAAH)Radiometric - رادیومتریTime-dependent inhibition - مهار وابسته به زمانBiomarkers - نشانگر زیستی یا بیومارکر
موضوعات مرتبط
مهندسی و علوم پایه
شیمی
شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
![عکس صفحه اول مقاله: High-performance liquid chromatography-tandem mass spectrometry assay of fatty acid amide hydrolase (FAAH) in blood: FAAH inhibition as clinical biomarker High-performance liquid chromatography-tandem mass spectrometry assay of fatty acid amide hydrolase (FAAH) in blood: FAAH inhibition as clinical biomarker](/preview/png/10533712.png)
چکیده انگلیسی
Fatty acid amide hydrolase (FAAH) is one of the main enzymes responsible for the degradation of the endocannabinoid anandamide (N-arachidonoylethanolamine, AEA). FAAH inhibitors may be useful in treating many disorders involving inflammation and pain. Although brain FAAH may be the relevant target for inhibition, rat studies show a correlation between blood and brain FAAH inhibition, allowing blood FAAH activity to be used as a target biomarker. Building on experience with a rat leukocyte FAAH activity assay using [3H]AEA, we have developed a human leukocyte assay using stably labeled [2H4]AEA as substrate. The deuterium-labeled ethanolamine reaction product ([2H4]EA) was analyzed by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in the positive electrospray ionization (ESI) mode. The response for [2H4]EA was linear from 10 nM to 10 μM, and the analysis time was less than 6 min/sample. Results using the [2H4]AEA and HPLC-MS/MS method agreed well with those obtained using the [3H]AEA radiometric assay. In addition to using a nonradioactive substrate, the HPLC-MS/MS method had increased sensitivity with lower background. Importantly, the assay preserved partial FAAH inhibition resulting from ex vivo treatment with a time-dependent irreversible inhibitor, suggesting its utility with clinical samples. The assay has been used to profile the successful inhibition of FAAH in recent clinical trials.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 421, Issue 2, 15 February 2012, Pages 556-565
Journal: Analytical Biochemistry - Volume 421, Issue 2, 15 February 2012, Pages 556-565
نویسندگان
Udeni Yapa, Jeffery J. Prusakiewicz, Ann D. Wrightstone, Lori J. Christine, Joe Palandra, Elizabeth Groeber, Arthur J. Wittwer,