Fast quantification of short chain fatty acids and ketone bodies by liquid chromatography-tandem mass spectrometry after facile derivatization coupled with liquid-liquid extraction

A rapid and convenient derivatization method for quantitation of short‐chain fatty acids in human feces by ultra‐performance liquid chromatography/tandem mass spectrometry

“…The total run time (5.5 min) of the LC/MS/MS analysis was shorter than or similar to that of other methods for derivatives of 2‐NPH (61 min), 3‐NPH (12 min), O‐BHA (7.5 min or 5.3 min), 2‐bromoacetophenone (18 min), and Dns‐PP (39 min).…”

“…Acetate interference was previously reported, preventing accurate quantitation. We also determined that the peak height of the acetic acid derivative was 7.3 × 10 5 counts in the double blank sample (no analyte, no IS, Figure S1A, supporting information).…”

“…The final optimum conditions determined for derivatization were a reaction time of 20 min at room temperature. The reaction conditions (room temperature for 20 min) were relatively mild and fast compared with those for 2‐NPH (60°C for 60 min), 3‐NPH (40°C for 30 min), O‐BHA (37°C for 20 min, or 25°C for 60 min), 2‐bromoacetophenone (40°C for 20 min), and Dns‐PP (37°C for 150 min). Therefore, the unstable SCFAs can quickly be converted with AMQ into stable derivatized products.…”

“…Research studies on several methods have been published to improve the chromatographic and MS behavior of SCFAs using liquid chromatography/tandem mass spectrometry (LC/MS/MS) through various chemical derivatization reactions. The following derivatization reagents have been reported: 2‐nitrophenylhydrazine (2‐NPH), 3‐nitrophenylhydrazine (3‐NPH), O ‐benzylhydroxylamine (O‐BHA), 2‐bromoacetophenone, and 5‐(dimethylamino)naphthalene‐1‐sulfonyl piperazine (Dns‐PP) . The aforementioned derivatization reactions required relatively high temperature, a long reaction time, complex derivatization steps, or a long analysis time in LC/MS/MS.…”

Fast quantification of short‐chain fatty acids in rat plasma by gas chromatography

“…Gas chromatography (GC) combined with polymerase chain reaction (Wang et al., 2017), liquid chromatography‐tandem mass spectrometry (Zeng & Cao 2018), gas chromatography/mass spectrometry (GC/MS) (Mulat & Feilberg 2015; Amer et al., 2015), Gas chromatography‐flame ionization detection (GC‐FID) (Zhang, Wang & Liu 2015), and the solid‐phase micro extraction (SPME)‐GC‐MS (Douny et al., 2019) methods have been used either directly or after derivatization to analyze SCFAs. Prior to GC analysis, the sample preparation processes as described in the previous studies commonly included solvent extraction, phase separation, evaporation to complete dryness, saponification under heat, and the addition of internal standards (Gao et al., 2018; Lu et al., 2018).…”