Development of an LC-MS Method for the Quantitative Determination of Six Organic Acids Produced by Bacterial Fermentation In Vitro and In Vivo
文献类型： 外文期刊
作者： Wang, Jing 1 ; Tang, Xuxiao 1 ; Qu, Chao 1 ; Zhang, Yingzhong 1 ; Zhang, Chaoqun 2 ; Chu, Qiulu 3 ; Wang, Hao 4 ;
作者机构： 1.Guangdong Acad Forestry, Guangdong Prov Key Lab Silviculture Protect & Util, Guangzhou 510520, Peoples R China
2.South China Agr Univ, Coll Mat & Energy, Key Lab Biobased Mat & Energy, Minist Educ, Guangzhou 510642, Peoples R China
3.Nanjing Forestry Univ, Coll Light Ind & Food Engn, Nanjing 210037, Peoples R China
4.Jiangsu Acad Agr Sci, Inst Vet Med, Key Lab Vet Biol Engn & Technol, Minist Agr & Rural Affairs, Nanjing 210014, Peoples R China
关键词： organic acids; extraction; LC-MS; quantification; in vitro; in vivo
期刊名称：PROCESSES （ 影响因子：2.8； 五年影响因子：3.1 ）
ISSN：
年卷期： 2025 年 13 卷 3 期
页码：
收录情况： SCI
摘要： Due to the strong polarity and volatility of organic acids, there is often a lack of effective quantitative methods for organic acids in vivo or in vitro. This study introduced a robust and scientifically validated LC-MS methodology for quantitatively analyzing six organic acids, encompassing five short-chain fatty acids and lactate, observed in in vitro fermentation and human fecal samples. This method was able to achieve precise quantification through the monitoring of mass-to-charge (m/z) ratios of the deprotonated negative ions. After optimization, a 2:1 (v/v) dichloromethane/acetonitrile mixture was utilized to extract the crude acid mix from fermentation or fecal samples. Subsequently, the organic acids were isolated using 3 mL of 3.5 M ammonium hydroxide solution from mixtures. The calibration curves for six organic acids demonstrated linearity with R2 > 0.991 across the concentration ranges of 0.01-5.0 or 0.01-20 mM. The coefficient of variation and accuracy were 2-13% and 95-128%, respectively, which exhibited good precision and sensitivity. Accurate quantification of organic acids would aid in metabolic analysis in bacterial culture supernatants and human fecal matter.
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