Short-chain fatty acids (SCFAs) are major gut microbiota-derived metabolites, which can reshape the intestine and regulate gut immunity. The application of conventional GC methods has been hampered for quantifying low-concentrated SCFAs, such as in serum, saliva, and digesta of germ-free animals. Herein, we established a LC–MS method to quantify SCFAs after 5-(dimethylamino)-1-carbohydrazide-isoquinoline (DMAQ) derivatization. The DMAQ derivatization significantly enhanced the detection sensitivity and improved separation of SCFAs. 2-methylbutyric acid and 3-methylbutyric acid were separately quantitated. Moreover, the matrix effect was diminished using DMAQ-13C/15N-tagged SCFAs as internal standards. The established quantitation method was successfully applied in the analysis of plasma and cecum digesta collected from neonatal piglets, revealing that significant increases in biological SCFA contents in cecum digesta were closely related to the variation of gut microbial diversity. The established quantitation method is capable of sensitively and comprehensively quantifying SCFAs that may provide insights into underlying gut-microbiota functions.
Ruyan Hou,Mengmeng Tong,Wanjun Gao,Lei Wang,Tianxi Yang,Lili He
To investigate the degradation and penetration behaviors of the organophosphate insecticide dimethoate applied on spinach leaves, in situ SERS and LC-MS methods were used to detect dimethoate residue on&in spinach leaves picked on different days after treatment (DAT). The SERS and LC-MS methods determined that the dimethoate degradation rate followed first-order kinetics, with a half-life of 3.56 or 4.13 days, depending on the respective method. The correlation coefficient of quantification value of these two methods was 0.9562. With the SERS method, we detected most of the penetrated dimethoate in the depth of 60–110 μm. With the LC-MS method, we detected dimethoate up to 0.17 ng in leaves and that the dimethoate on:in ratio of spinach leaves ranged from 562.25 on 0 DAT to 5.23 on 14 DAT. The combination of these two methods facilitated a better understanding of the behavior and biological fate of pesticides in a complex biological system.