Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S]
We present a method using a combination of enzymatic deconjugation and targeted LC-multiple reaction monitoring (MRM)-MS analysis for analyzing all common bile acids (BAs) in piglet urine, and in particular, for detecting conjugated BAs either in the absence of their standards, or when present in lo...
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| Format: | Article |
| Language: | English |
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Elsevier
2016-10-01
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| Series: | Journal of Lipid Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S0022227520354006 |
| _version_ | 1818412095166742528 |
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| author | Nianbai Fang Shanggong Yu Sean H. Adams MartinJ.J. Ronis Thomas M. Badger |
| author_facet | Nianbai Fang Shanggong Yu Sean H. Adams MartinJ.J. Ronis Thomas M. Badger |
| author_sort | Nianbai Fang |
| collection | DOAJ |
| description | We present a method using a combination of enzymatic deconjugation and targeted LC-multiple reaction monitoring (MRM)-MS analysis for analyzing all common bile acids (BAs) in piglet urine, and in particular, for detecting conjugated BAs either in the absence of their standards, or when present in low concentrations. Initially, before enzymatic deconjugation, 19 unconjugated BAs (FBAs) were detected where the total concentration of the detected FBAs was 9.90 μmol/l. Sixty-seven conjugated BAs were identified by LC-MRM-MS analysis before and after enzymatic deconjugation. Four enzymatic assays were used to deconjugate the BA conjugates. FBAs in urine after cholylglycine hydrolase/sulfatase treatment were 33.40 μmol/l, indicating the urinary BAs were comprised of 29.75% FBAs and 70.25% conjugated BAs in single and multiple conjugated forms. For the conjugates in single form, released FBAs from cholylglycine hydrolase deconjugation indicated that the conjugates with amino acids were 14.54% of urinary BAs, 16.27% glycosidic conjugates were found by β-glucuronidase treatment, and sulfatase with glucuronidase inhibitor treatment liberated FBAs that constituted 16.67% of urinary BAs. Notably, chenodeoxycholic acid (CDCA) was initially detected only in trace amounts in urine, but was found at significant levels after the enzymatic assays above. These results support that CDCA is a precursor of γ-muricholic acid in BA biosynthesis in piglets. |
| first_indexed | 2024-12-14T10:41:51Z |
| format | Article |
| id | doaj.art-808aef7f449a42c581789648e610c15f |
| institution | Directory Open Access Journal |
| issn | 0022-2275 |
| language | English |
| last_indexed | 2024-12-14T10:41:51Z |
| publishDate | 2016-10-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Lipid Research |
| spelling | doaj.art-808aef7f449a42c581789648e610c15f2022-12-21T23:05:39ZengElsevierJournal of Lipid Research0022-22752016-10-01571019171933Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S]Nianbai Fang0Shanggong Yu1Sean H. Adams2MartinJ.J. Ronis3Thomas M. Badger4Arkansas Children's Nutrition Center University of Arkansas for Medical Sciences, Little Rock, AR; Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, AR; To whom correspondence should be addressed. (N.F.); (T.M.B.)Arkansas Children's Nutrition Center University of Arkansas for Medical Sciences, Little Rock, ARArkansas Children's Nutrition Center University of Arkansas for Medical Sciences, Little Rock, AR; Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, ARArkansas Children's Nutrition Center University of Arkansas for Medical Sciences, Little Rock, AR; Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, ARArkansas Children's Nutrition Center University of Arkansas for Medical Sciences, Little Rock, AR; Department of Pediatrics, University of Arkansas for Medical Sciences, Little Rock, AR; To whom correspondence should be addressed. (N.F.); (T.M.B.)We present a method using a combination of enzymatic deconjugation and targeted LC-multiple reaction monitoring (MRM)-MS analysis for analyzing all common bile acids (BAs) in piglet urine, and in particular, for detecting conjugated BAs either in the absence of their standards, or when present in low concentrations. Initially, before enzymatic deconjugation, 19 unconjugated BAs (FBAs) were detected where the total concentration of the detected FBAs was 9.90 μmol/l. Sixty-seven conjugated BAs were identified by LC-MRM-MS analysis before and after enzymatic deconjugation. Four enzymatic assays were used to deconjugate the BA conjugates. FBAs in urine after cholylglycine hydrolase/sulfatase treatment were 33.40 μmol/l, indicating the urinary BAs were comprised of 29.75% FBAs and 70.25% conjugated BAs in single and multiple conjugated forms. For the conjugates in single form, released FBAs from cholylglycine hydrolase deconjugation indicated that the conjugates with amino acids were 14.54% of urinary BAs, 16.27% glycosidic conjugates were found by β-glucuronidase treatment, and sulfatase with glucuronidase inhibitor treatment liberated FBAs that constituted 16.67% of urinary BAs. Notably, chenodeoxycholic acid (CDCA) was initially detected only in trace amounts in urine, but was found at significant levels after the enzymatic assays above. These results support that CDCA is a precursor of γ-muricholic acid in BA biosynthesis in piglets.http://www.sciencedirect.com/science/article/pii/S0022227520354006liquid chromatography-multiple reaction monitoring-mass spectrometryenzymatic deconjugationpiglet urinebile acid conjugates |
| spellingShingle | Nianbai Fang Shanggong Yu Sean H. Adams MartinJ.J. Ronis Thomas M. Badger Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] Journal of Lipid Research liquid chromatography-multiple reaction monitoring-mass spectrometry enzymatic deconjugation piglet urine bile acid conjugates |
| title | Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] |
| title_full | Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] |
| title_fullStr | Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] |
| title_full_unstemmed | Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] |
| title_short | Profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted LC-MRM-MS[S] |
| title_sort | profiling of urinary bile acids in piglets by a combination of enzymatic deconjugation and targeted lc mrm ms s |
| topic | liquid chromatography-multiple reaction monitoring-mass spectrometry enzymatic deconjugation piglet urine bile acid conjugates |
| url | http://www.sciencedirect.com/science/article/pii/S0022227520354006 |
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