A Facile Profiling Method of Short Chain Fatty Acids Using Liquid Chromatography-Mass Spectrometry
Abstract
:1. Introduction
2. Materials and Methods
2.1. Materials
2.2. Sample Preparation
2.3. Biological Samples
2.4. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS)
2.5. Method Validation
3. Results and Discussion
3.1. Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) of Derivatized Short Chain Fatty Acids
3.2. Optimization of the Derivatization Reaction and Method Validation
3.3. Profiling of SCFAs in Biological Samples
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
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Compound | DP | EP | MRM Transitions | IS | Calibration Curve | LOQ, μM (ng/mL) | Intraday (10 μM) | Interday (10 μM) | ||
---|---|---|---|---|---|---|---|---|---|---|
(V) | (V) | Q1 | Q3 | Calibration Range (μM) | R2 | RSD, % (Accuracy, %) | RSD, % (Accuracy, %) | |||
Acetic acid | 70 | 8 | 252 | 210 | A | 1–1000 | 1 | 1.03 (61.8) | 6.66 (98.5) | 14.9 (103) |
Propionic acid | 30 | 10 | 266 | 210 | B | 0.1–1000 | 0.999 | 0.293 (21.7) | 0.909 (110) | 15.1 (97.4) |
Isobutyric acid | 70 | 6 | 280 | 210 | C | 0.1–1000 | 0.999 | 0.450 (39.6) | 1.05 (110) | 7.97 (93.2) |
Butyric acid | 70 | 6 | 280 | 210 | C | 0.1–1000 | 1 | 0.158 (13.9) | 1.72 (116) | 13.4 (97.0) |
2,2-dimethylpropionic acid | 50 | 10 | 294 | 210 | D | 0.1–100 | 0.989 | 0.470 (47.9) | 1.75 (110) | 11.3 (86.1) |
2-Methylbutyric acid | 50 | 10 | 294 | 210 | D | 0.1–500 | 0.998 | 0.773 (78.8) | 1.50 (102) | 8.87 (86.9) |
Isovaleric acid | 50 | 10 | 294 | 210 | D | 0.1–100 | 0.999 | 0.624 (63.6) | 2.11 (103) | 9.09 (91.4) |
Valeric acid | 50 | 10 | 294 | 210 | D | 0.1–1000 | 0.997 | 0.451 (46.0) | 0.475 (101) | 6.00 (87.0) |
2,2-Dimethylbutyric acid | 70 | 10 | 308 | 210 | E | 0.1–100 | 0.997 | 0.430 (49.9) | 7.67 (116) | 11.6 (85.7) |
2-Ethylbutyric acid, | 70 | 10 | 308 | 210 | E | 0.1–100 | 0.999 | 0.546 (63.3) | 4.83 (108) | 12.2 (96.0) |
2-Methylvaleric acid | 70 | 10 | 308 | 210 | E | 0.1–500 | 0.983 | 0.773 (89.7) | 3.38 (112) | 4.49 (105) |
Caproic acid | 70 | 10 | 308 | 210 | E | 0.1–1000 | 0.994 | 0.609 (70.6) | 1.66 (89.8) | 4.74 (89.3) |
Acetic acid-2,2,2-d3 (A) | 70 | 6 | 255 | 211 | - | - | - | - | - | - |
Propionic acid-d6 (B) | 30 | 8 | 272 | 211 | - | - | - | - | - | - |
Butyric acid- d7 (C) | 70 | 8 | 287 | 211 | - | - | - | - | - | - |
Valeric acid-2,2,3,3-d4 (D) | 50 | 8 | 298 | 211 | - | - | - | - | - | - |
Caproic-5,5,6,6,6-d5 (E) | 70 | 8 | 313 | 210 | - | - | - | - | - | - |
Mouse Feces (nmol/mg) | Mouse plasma (μM) | Human EBC (μM) | |||||||
---|---|---|---|---|---|---|---|---|---|
Average | Intraday | Interday | Average | Intraday | Interday | Average | Intraday | Interday | |
RSD (%) | RSD (%) | RSD (%) | RSD (%) | RSD (%) | RSD (%) | ||||
Acetic acid | 19.5 | 3.25 | 8.04 | 77.4 | 1.61 | 8.03 | 66.4 | 1.27 | 6.02 |
Propionic acid | 3.54 | 0.866 | 2.88 | 8.08 | 3.1 | 2.27 | 13.4 | 1.17 | 2.41 |
Isobutyric acid | 0.802 | 4.87 | 9.23 | 31.1 | 7.37 | 9.74 | 2.43 | 5.2 | 6.15 |
Butyric acid | 0.0934 | 8.13 | 11.2 | 20.7 | 5.46 | 8.98 | 2.79 | 6.01 | 2.28 |
2,2-Dimethylpropionic acid | ND | - | - | 35.9 | 5.17 | 9.73 | ND | - | - |
2-Methylbutyric acid | 0.645 | 1.12 | 10 | 14.3 | 6.54 | 12.5 | ND | - | - |
Isovaleric acid | 0.379 | 1.29 | 8.84 | 9.92 | 6.95 | 18 | ND | - | - |
Valeric acid | 0.224 | 1.64 | 4.53 | 6.16 | 10.4 | 11.5 | ND | - | - |
2,2-Dimethylbutyric acid | ND | - | - | ND | - | - | ND | - | - |
2-Ethylbutyric acid | ND | - | - | 0.651 | 7.36 | 11.1 | ND | - | - |
2-Methylvaleric acid | ND | - | - | 2.09 | 6.07 | 2.01 | ND | - | - |
Caproic acid | 0.0125 | 8.74 | 9.61 | 0.711 | 6.66 | 8.95 | ND | - | - |
HFD Mouse Feces, (nmol/mg) | LFD Mouse Feces, (nmol/mg) | t Test | |||
---|---|---|---|---|---|
Average | STD | Average | STD | ||
Acetic acid | 16.4 | 0.659 | 19.5 | 1.57 | * |
Propionic acid | 2.99 | 0.118 | 3.54 | 0.102 | ** |
Isobutyric acid | 0.416 | 0.0655 | 0.802 | 0.074 | ** |
Butyric acid | 0.821 | 0.0352 | 0.0934 | 0.0105 | ** |
2,2-dimethylpropionic acid | ND | - | ND | - | |
2-Methylbutyric acid | 0.534 | 0.0503 | 0.645 | 0.0646 | |
Isovaleric acid | 0.675 | 0.0046 | 0.379 | 0.0335 | ** |
Valeric acid | 1.389 | 0.0698 | 0.223 | 0.0101 | ** |
2,2-Dimethylbutyric acid | ND | - | ND | - | |
2-Ethylbutyric acid | ND | - | ND | - | |
2-Methylvaleric acid | ND | - | ND | - | |
Caproic acid | 0.00478 | 0.000357 | 0.0125 | 0.0012 | ** |
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Song, H.E.; Lee, H.Y.; Kim, S.J.; Back, S.H.; Yoo, H.J. A Facile Profiling Method of Short Chain Fatty Acids Using Liquid Chromatography-Mass Spectrometry. Metabolites 2019, 9, 173. https://doi.org/10.3390/metabo9090173
Song HE, Lee HY, Kim SJ, Back SH, Yoo HJ. A Facile Profiling Method of Short Chain Fatty Acids Using Liquid Chromatography-Mass Spectrometry. Metabolites. 2019; 9(9):173. https://doi.org/10.3390/metabo9090173
Chicago/Turabian StyleSong, Ha Eun, Hyo Yeong Lee, Su Jung Kim, Sung Hoon Back, and Hyun Ju Yoo. 2019. "A Facile Profiling Method of Short Chain Fatty Acids Using Liquid Chromatography-Mass Spectrometry" Metabolites 9, no. 9: 173. https://doi.org/10.3390/metabo9090173