Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS
Urine organic acid contains water-soluble metabolites and/or metabolites—derived from sugars, amino acids, lipids, vitamins, and drugs—which can reveal a human’s physiological condition. These urine organic acids—hippuric acid, benzoic acid, phenylacetic acid, phenylpropionic acid, 4-hydroxybenzoic...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-08-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/27/17/5363 |
| _version_ | 1797494251100045312 |
|---|---|
| author | Yu-Tsung Lee Sui-Qing Huang Ching-Hao Lin Li-Heng Pao Chun-Hui Chiu |
| author_facet | Yu-Tsung Lee Sui-Qing Huang Ching-Hao Lin Li-Heng Pao Chun-Hui Chiu |
| author_sort | Yu-Tsung Lee |
| collection | DOAJ |
| description | Urine organic acid contains water-soluble metabolites and/or metabolites—derived from sugars, amino acids, lipids, vitamins, and drugs—which can reveal a human’s physiological condition. These urine organic acids—hippuric acid, benzoic acid, phenylacetic acid, phenylpropionic acid, 4-hydroxybenzoic acid, 4-hydroxyphenyl acetic acid, 3-hydroxyphenylpropionic acid, 3,4-dihydroxyphenyl propionic acid, and 3-indoleacetic acid—were the eligible candidates for the dysbiosis of gut microbiota. The aim of this proposal was to develop and to validate a liquid chromatography–tandem mass spectrometry (LC-MS/MS) bioanalysis method for the nine organic acids in human urine. Stable-labeled isotope standard (creatinine-d<sub>3</sub>) and acetonitrile were added to the urine sample. The supernatant was diluted with deionized water and injected into LC-MS/MS. This method was validated with high selectivity for the urine sample, a low limit of quantification (10–40 ng/mL), good linearity (<i>r</i> > 0.995), high accuracy (85.8–109.7%), and high precision (1.4–13.3%). This method simultaneously analyzed creatinine in urine, which calibrates metabolic rate between different individuals. Validation has been completed for this method; as such, it could possibly be applied to the study of gut microbiota clinically. |
| first_indexed | 2024-03-10T01:31:40Z |
| format | Article |
| id | doaj.art-6e3cf48c86e74088a9e8084bbf13ff7c |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T01:31:40Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-6e3cf48c86e74088a9e8084bbf13ff7c2023-11-23T13:40:45ZengMDPI AGMolecules1420-30492022-08-012717536310.3390/molecules27175363Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MSYu-Tsung Lee0Sui-Qing Huang1Ching-Hao Lin2Li-Heng Pao3Chun-Hui Chiu4Graduate Institute of Health Industry and Technology, Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, TaiwanGraduate Institute of Health Industry and Technology, Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, TaiwanGraduate Institute of Health Industry and Technology, Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, TaiwanGraduate Institute of Health Industry and Technology, Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, TaiwanGraduate Institute of Health Industry and Technology, Research Center for Food and Cosmetic Safety, College of Human Ecology, Chang Gung University of Science and Technology, Taoyuan 33303, TaiwanUrine organic acid contains water-soluble metabolites and/or metabolites—derived from sugars, amino acids, lipids, vitamins, and drugs—which can reveal a human’s physiological condition. These urine organic acids—hippuric acid, benzoic acid, phenylacetic acid, phenylpropionic acid, 4-hydroxybenzoic acid, 4-hydroxyphenyl acetic acid, 3-hydroxyphenylpropionic acid, 3,4-dihydroxyphenyl propionic acid, and 3-indoleacetic acid—were the eligible candidates for the dysbiosis of gut microbiota. The aim of this proposal was to develop and to validate a liquid chromatography–tandem mass spectrometry (LC-MS/MS) bioanalysis method for the nine organic acids in human urine. Stable-labeled isotope standard (creatinine-d<sub>3</sub>) and acetonitrile were added to the urine sample. The supernatant was diluted with deionized water and injected into LC-MS/MS. This method was validated with high selectivity for the urine sample, a low limit of quantification (10–40 ng/mL), good linearity (<i>r</i> > 0.995), high accuracy (85.8–109.7%), and high precision (1.4–13.3%). This method simultaneously analyzed creatinine in urine, which calibrates metabolic rate between different individuals. Validation has been completed for this method; as such, it could possibly be applied to the study of gut microbiota clinically.https://www.mdpi.com/1420-3049/27/17/5363gut microbiotaLC-MS/MSorganic acidhuman urine |
| spellingShingle | Yu-Tsung Lee Sui-Qing Huang Ching-Hao Lin Li-Heng Pao Chun-Hui Chiu Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS Molecules gut microbiota LC-MS/MS organic acid human urine |
| title | Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS |
| title_full | Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS |
| title_fullStr | Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS |
| title_full_unstemmed | Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS |
| title_short | Quantification of Gut Microbiota Dysbiosis-Related Organic Acids in Human Urine Using LC-MS/MS |
| title_sort | quantification of gut microbiota dysbiosis related organic acids in human urine using lc ms ms |
| topic | gut microbiota LC-MS/MS organic acid human urine |
| url | https://www.mdpi.com/1420-3049/27/17/5363 |
| work_keys_str_mv | AT yutsunglee quantificationofgutmicrobiotadysbiosisrelatedorganicacidsinhumanurineusinglcmsms AT suiqinghuang quantificationofgutmicrobiotadysbiosisrelatedorganicacidsinhumanurineusinglcmsms AT chinghaolin quantificationofgutmicrobiotadysbiosisrelatedorganicacidsinhumanurineusinglcmsms AT lihengpao quantificationofgutmicrobiotadysbiosisrelatedorganicacidsinhumanurineusinglcmsms AT chunhuichiu quantificationofgutmicrobiotadysbiosisrelatedorganicacidsinhumanurineusinglcmsms |