Targeted profiling of circulating and hepatic bile acids in human, mouse, and rat using a UPLC-MRM-MS-validated method

García-Cañaveras, Juan Carlos; Donato, M. Teresa; Castell, José V.; Lahoz, Agustín

doi:10.1194/jlr.d028803

Cited by 236 publications

(214 citation statements)

References 39 publications

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“…As can be seen, intraday precision and accuracy of the 57 BAs were consistent with inter-day precision and accuracy for the QC1, QC2, QC3 and QC4 compared to previous work that showed significant decreased of the inter-day accuracies. 27 Figure 4. Accuracy distribution of the QC1 (10 nM), QC2 (50 nM), QC3 (100 nM), QC4 (750nM) on intra-and inter-day.…”

Section: Intra-and Inter Day Precision and Accuracymentioning

confidence: 99%

“…Similar results have been found in previous work for QC1 where values were approaching the saturation limit. 27 Conversely, QC2, QC3 and QC4, which benefit from being far from the noise and the saturation level,…”

Section: Intra-and Inter Day Precision and Accuracymentioning

confidence: 99%

“…[15][16][17] Within the past decade, LC-MS has been heavily utilized for the separation and detection of BAs in human and animal model biofluid samples. 9,[18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] The most comprehensive of these methods have a relatively long analytical cycle (> 30 minutes), precluding rapid sample analysis. 21,25,28,30,34 On the other hand, the most rapid methods (< 10 minutes) have limited BA coverage.…”

mentioning

confidence: 99%

“…19,33 The majority of separations have analytical times greater than 20 minutes and intermediate level of BA species coverage (ranging from 11 to 32 species monitored). 9,[18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][33][34] Very few LC-MS methods report coverage of tertiary BAs such as sulfate conjugates which have recently been implicated in the important role of the gut microbiota in human metabolism. 9,32,[34][35][36] The analytical foundation of these methods is reversed-phase separation utilizing a C 18 stationary phase, which is suitable for retention and separation of the diverse range of hydrophobicity present among BAs.…”

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confidence: 99%

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Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

et al. 2015

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Bile acids are important end products of cholesterol metabolism. While they have been identified as key factors in lipid emulsification and absorption due to their detergent properties, bile acids have also been shown to act as signaling molecules and intermediates between the host and the gut microbiota. To further the investigation of bile acid functions in humans, an advanced platform for high throughput analysis is essential. Herein, we describe the development and application of a 15 min UPLC procedure for the separation of bile acid species from human biofluid samples requiring minimal sample preparation. High resolution time-of-flight mass spectrometry was applied for profiling applications, elucidating rich bile acid profiles in both normal and disease state plasma. In parallel, a second mode of detection was developed utilizing tandem mass spectrometry for sensitive and quantitative targeted analysis of 145 bile acid (BA) species including primary, secondary, and tertiary bile acids. The latter system was validated by testing the linearity (lower limit of quantification, LLOQ, 0.25?10 nM and upper limit of quantification, ULOQ, 2.5?5 ?M), precision (?6.5%), and accuracy (81.2?118.9%) on inter- and intraday analysis achieving good recovery of bile acids (serum/plasma 88% and urine 93%). The ultra performance liquid chromatography?mass spectrometry (UPLC-MS)/MS targeted method was successfully applied to plasma, serum, and urine samples in order to compare the bile acid pool compositional difference between preprandial and postprandial states, demonstrating the utility of such analysis on human biofluids

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Section: Intra-and Inter Day Precision and Accuracymentioning

confidence: 99%

Section: Intra-and Inter Day Precision and Accuracymentioning

confidence: 99%

mentioning

confidence: 99%

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confidence: 99%

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Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

et al. 2015

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“…10 Although there is a consensus in the literature regarding the use of multiple reaction monitoring (MRM) for the quantication of taurineand glycine-conjugated BAs, MRM has not yet been widely used for quantication of unconjugated BAs. 11,[13][14][15][16][17][18][19][20][21][22][23][24][25] In the present study, we applied specic fragmentation using MRM for both conjugated and unconjugated BAs in order to increase the specicity of detection and sensitivity for quantication of BAs in complex biological matrices such as serum and plasma.…”

Section: Introductionmentioning

confidence: 99%

Quantification of multiple bile acids in uninephrectomized rats using ultra-performance liquid chromatography-tandem mass spectrometry

et al. 2013

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In order to study the roles of individual BAs and due to limited blood sample volumes available from experimental animals, improved methods for the simultaneous quantification of multiple BAs are needed. We developed and validated an ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method for the quantification of 24 BAs, including 11 unconjugated, 6 glycine-conjugated and 7 taurine-conjugated BAs, in 50 mL of rat serum or plasma. The UPLC-MS/MS method, operated in negative and positive ion mode, allows quantification of BAs using multiplereaction monitoring (MRM), with specific fragmentation of BAs. The method showed acceptable intraand inter-day accuracy, precision, extraction recovery and high sensitivity, with a lower limit of quantification (LLOQ) in the pM range for several taurine-conjugated BAs. We applied the established method to investigate potential time-dependent changes of BAs in plasma from sham-operated and uninephrectomized male Sprague-Dawley rats. The levels of several primary and secondary BAs were transiently elevated one week after uninephrectomy, followed by normalization thereafter. In contrast, several conjugated BAs were slightly increased after the second week post-surgery. The established UPLC-MS/MS method, employing specific fragmentation of free and conjugated BAs by MRM, allows the simultaneous quantification of multiple BAs in 50 mL serum or plasma samples, and can be used to assess BA profiles in patho-physiological situations.

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Mechanistic Modeling of Drug‐Induced Liver Injury (DILI)

Yang

Woodhead

Shoda

et al. 2015

Methods and Principles in Medicinal Chemistry

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Targeted profiling of circulating and hepatic bile acids in human, mouse, and rat using a UPLC-MRM-MS-validated method

Cited by 236 publications

References 39 publications

Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

Bile Acid Profiling and Quantification in Biofluids Using Ultra-Performance Liquid Chromatography Tandem Mass Spectrometry

Quantification of multiple bile acids in uninephrectomized rats using ultra-performance liquid chromatography-tandem mass spectrometry

Mechanistic Modeling of Drug‐Induced Liver Injury (DILI)

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