کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1164267 1490978 2015 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
An isotope-labeled chemical derivatization method for the quantitation of short-chain fatty acids in human feces by liquid chromatography–tandem mass spectrometry
ترجمه فارسی عنوان
یک روش مشتق شده شیمیایی با برچسب ایزوتوپ برای کم کردن اسید چرب کوتاه مدت در مدفوع انسان با استفاده از اسپکترومتری جرمی کروماتوگرافی
کلمات کلیدی
اسید چرب کوتاه مدت، نشانه گذاری ایزوتوپ، مشتقات شیمیایی، 3-نیترفانیل هیدرازین، طیف سنجی جرمی کروماتوگرافی مایع با عملکرد فوق العاده بالا - چندتایی واکنش سنجی جرمی
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• 3-Nitrophenylhydrazine was used to derivatize short-chain fatty acids (SCFAs) for LC-MS/MS.
• 13C6 analogues were produced for use as isotope-labeled internal standards.
• Isotope-labeled standards compensate for ESI matrix effects in LC-MS/MS.
• Femtomolar sensitivities and 93–108% quantitation accuracy were achieved for human fecal SCFAs.

Short-chain fatty acids (SCFAs) are produced by anaerobic gut microbiota in the large bowel. Qualitative and quantitative measurements of SCFAs in the intestinal tract and the fecal samples are important to understand the complex interplay between diet, gut microbiota and host metabolism homeostasis. To develop a new LC-MS/MS method for sensitive and reliable analysis of SCFAs in human fecal samples, 3-nitrophenylhydrazine (3NPH) was employed for pre-analytical derivatization to convert ten C2–C6 SCFAs to their 3-nitrophenylhydrazones under a single set of optimized reaction conditions and without the need of reaction quenching. The derivatives showed excellent in-solution chemical stability. They were separated on a reversed-phase C18 column and quantitated by negative-ion electrospray ionization – multiple-reaction monitoring (MRM)/MS. To achieve accurate quantitation, the stable isotope-labeled versions of the derivatives were synthesized in a single reaction vessel from 13C6-3NPH, and were used as internal standard to compensate for the matrix effects in ESI. Method validation showed on-column limits of detection and quantitation over the range from low to high femtomoles for the ten SCFAs, and the intra-day and inter-day precision for determination of nine of the ten SCFAs in human fecal samples was ≤8.8% (n = 6). The quantitation accuracy ranged from 93.1% to 108.4% (CVs ≤ 4.6%, n = 6). This method was used to determine the SCFA concentrations and compositions in six human fecal samples. One of the six samples, which was collected from a clinically diagnosed type 2 diabetes patient showed a significantly high molar ratio of branch-chain SCFAs to straight-chain SCFAs than the others. In summary, this work provides a new LC-MS/MS method for precise and accurate quantitation of SCFAs in human feces.

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ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytica Chimica Acta - Volume 854, 7 January 2015, Pages 86–94
نویسندگان
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