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A sensitive method for the quantification of short-chain fatty acids by benzyl chloroformate derivatization combined with GC-MS.
Analyst ( IF 3.6 ) Pub Date : 2020/02/10 , DOI: 10.1039/d0an00005a Menghan Li 1 , Rongrong Zhu , Xiaoxia Song , Zhijun Wang , Hongbo Weng , Jianying Liang
Analyst ( IF 3.6 ) Pub Date : 2020/02/10 , DOI: 10.1039/d0an00005a Menghan Li 1 , Rongrong Zhu , Xiaoxia Song , Zhijun Wang , Hongbo Weng , Jianying Liang
Affiliation
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Short-chain fatty acids (SCFAs) were identified as critical markers in the diagnosis of chronic and metabolic diseases, but a sensitive and stable method to determine SCFAs in feces is a challenge for analysts due to the high volatility. Herein, a sensitive and accurate method to determine SCFAs adopting precolumn derivatization coupled with gas chromatography-mass spectrometry (GC-MS) has been developed. Benzyl chloroformate (BCF) was chosen as the reaction reagent and emulsified derivatization was applied to homogenize the reaction system. Higher sensitivity, wider application and satisfactory derivatization efficiency were obtained using the developed method. An excellent method validation showed a good linearity ranging from 0.9947 to 0.9998. At the same time, the intra-day and inter-day precision were achieved in the range of 0.56% to 13.07%. The lower limits of detection of all target analytes varied from 0.1 to 5 pg. The recovery ranged from 80.87% to 119.03%, and storage stability under three different conditions was also determined. This method was also successfully applied to the analysis of SCFAs in mice fecal samples to illustrate the significant differences between normal and type 2 diabetes mellitus mice.
中文翻译：
氯甲酸苄酯衍生化与GC-MS结合使用的定量短链脂肪酸的灵敏方法。
短链脂肪酸（SCFA）被确定为诊断慢性和代谢性疾病的关键标志，但是由于挥发性高，测定粪便中SCFA的灵敏且稳定的方法是分析人员面临的挑战。在此，开发了一种灵敏而准确的方法，即采用柱前衍生化结合气相色谱-质谱（GC-MS）测定SCFA。选择氯甲酸苄酯（BCF）作为反应试剂，并进行乳化衍生化反应体系。使用该方法获得了更高的灵敏度，更广泛的应用和令人满意的衍生化效率。出色的方法验证表明线性范围为0.9947至0.9998。同时，日内和日间精度达到0.56％至13.07％的范围。所有目标分析物的检测下限为0.1到5 pg。回收率从80.87％到119.03％，并且还确定了在三种不同条件下的储存稳定性。该方法还成功地应用于小鼠粪便样品中的SCFA分析，以说明正常小鼠和2型糖尿病小鼠之间的显着差异。
更新日期：2020-03-31
中文翻译：
氯甲酸苄酯衍生化与GC-MS结合使用的定量短链脂肪酸的灵敏方法。
短链脂肪酸（SCFA）被确定为诊断慢性和代谢性疾病的关键标志，但是由于挥发性高，测定粪便中SCFA的灵敏且稳定的方法是分析人员面临的挑战。在此，开发了一种灵敏而准确的方法，即采用柱前衍生化结合气相色谱-质谱（GC-MS）测定SCFA。选择氯甲酸苄酯（BCF）作为反应试剂，并进行乳化衍生化反应体系。使用该方法获得了更高的灵敏度，更广泛的应用和令人满意的衍生化效率。出色的方法验证表明线性范围为0.9947至0.9998。同时，日内和日间精度达到0.56％至13.07％的范围。所有目标分析物的检测下限为0.1到5 pg。回收率从80.87％到119.03％，并且还确定了在三种不同条件下的储存稳定性。该方法还成功地应用于小鼠粪便样品中的SCFA分析，以说明正常小鼠和2型糖尿病小鼠之间的显着差异。
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