scholarly journals Identification and Distribution of Sterols, Bile Acids, and Acylcarnitines by LC–MS/MS in Humans, Mice, and Pigs—A Qualitative Analysis

Metabolites ◽  
2022 ◽  
Vol 12 (1) ◽  
pp. 49
Author(s):  
Ambrin Farizah Babu ◽  
Ville Mikael Koistinen ◽  
Soile Turunen ◽  
Gloria Solano-Aguilar ◽  
Joseph F. Urban ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Bile Acid ◽  
Bile Acids ◽  
Biological Samples ◽  
Simultaneous Detection ◽  
Reversed Phase ◽  
Proximal Colon ◽  
Low Concentrations ◽  
Human Stool ◽  
Mouse Ileum

Sterols, bile acids, and acylcarnitines are key players in human metabolism. Precise annotations of these metabolites with mass spectrometry analytics are challenging because of the presence of several isomers and stereoisomers, variability in ionization, and their relatively low concentrations in biological samples. Herein, we present a sensitive and simple qualitative LC–MS/MS (liquid chromatography with tandem mass spectrometry) method by utilizing a set of pure chemical standards to facilitate the identification and distribution of sterols, bile acids, and acylcarnitines in biological samples including human stool and plasma; mouse ileum, cecum, jejunum content, duodenum content, and liver; and pig bile, proximal colon, cecum, heart, stool, and liver. With this method, we detected 24 sterol, 32 bile acid, and 27 acylcarnitine standards in one analysis that were separated within 13 min by reversed-phase chromatography. Further, we observed different sterol, bile acid, and acylcarnitine profiles for the different biological samples across the different species. The simultaneous detection and annotation of sterols, bile acids, and acylcarnitines from reference standards and biological samples with high precision represents a valuable tool for screening these metabolites in routine scientific research.

scholarly journals Polyphenol Consumption on Human Bile Acids Metabolism: Preliminary Data of Bile Acid Profiles in Human Biological Samples (P06-131-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Anqi Zhao ◽  
Xuhuiqun Zhang ◽  
Amandeep Sandhu ◽  
Indika Edirisinghe ◽  
Barbara Shukitt-Hale ◽  
...  
Keyword(s):  
Bile Acid ◽  
Qualitative Analysis ◽  
Bile Acids ◽  
Biological Samples ◽  
Urine Samples ◽  
Fragmentation Pattern ◽  
Plasma Samples ◽  
Potential Health ◽  
Human Bile ◽  
Freeze Dried

Abstract Objectives Bile acids (BAs) play a critical role in regulating human health through the activation of BAs receptor farnesoid X receptor (FXR) and membrane G protein coupled bile acid receptor-1 (TGR5). We aimed to develop methods to characterize BAs and their metabolites in human biological samples and characterize changes in BAs profile after chronic polyphenol consumption to help guide investigations on the potential health effects of polyphenols via BAs metabolism. Methods Plasma, fecal and urine samples from two human studies that included berry intake were used for developing qualitative analysis of BAs using ultra high-performance liquid chromatography (UHPLC) coupled with electrospray ionization quadrupole time of flight (QTOF). The compounds were identified based on the exact mass, fragmentation pattern, available reference standards and database search. To investigate the effect of chronic polyphenol consumption on BAs composition, pooled plasma samples (fasting and postprandial 2 h, n = 6) from a chronic (45 and 90 days) strawberry supplementation study (24 g freeze dried/day) with an older population were analyzed. Results Among 106 BAs and their metabolites which were tentatively identified in the samples used for method development, 70, 55, and 47 BAs species were characterized in plasma, feces and urine samples, respectively. The qualitative analysis of BAs in plasma samples from subjects following the strawberry consumption protocol detected 8 primary and 31 secondary BAs. After 90-days strawberry supplementation, two secondary BAs–glycolithocholic acid and 9(11), (5β)-cholenic acid-3α, 12α-diol were decreased to undetectable levels in the pooled fasting sample and the FRX/TGR5 agonists, including chenodeoxycholic acid, deoxycholic acid, cholic acid, glycodeoxycholic acid and taurocholic acid, showed increasing peak areas at 2 h postprandial compared to fasting. Conclusions The changes in BAs profiles in fasting and postprandial plasma samples after chronic strawberry feeding suggest that strawberry polyphenols may alter BAs metabolism and the FXR/TGR5 signaling. Funding Sources This work was funded by the California Strawberry Commission, USDA Intramural Funds and various donor funds to the Center for Nutrition Research, IIT.

A systems-level analysis of bile acids effects on rat colon epithelial cells

2021 ◽  
Author(s):  
Jonathan Yde ◽  
Qi Wu ◽  
Johan Borg ◽  
Robert A. Fenton ◽  
Hanne Bjerregaard Moeller
Keyword(s):  
Mass Spectrometry ◽  
Bile Acid ◽  
Epithelial Cells ◽  
Bile Acids ◽  
Rna Sequencing ◽  
Urine Volume ◽  
Rat Colon ◽  
Acid Transport ◽  
Bile Acid Transport ◽  
Underlying Mechanisms

Bile acid diarrhoea is a chronic condition caused by increased delivery of bile acids to the colon. The underlying mechanisms remain to be elucidated. To investigate genes involved in bile acid diarrhoea, systems-level analyses were employed on a rat bile acid diarrhoea model. Twelve male Wistar Munich rats, housed in metabolic cages, were fed either control or bile acid-mixed (1% w/w) diets for ten days. Food intake, water intake, urine volume, bodyweight and faecal output were monitored daily. After euthanasia, colonic epithelial cells were isolated using calcium-chelation and processed for systems-level analyses, i.e. RNA-sequencing transcriptomics and mass spectrometry proteomics. Bile acid-fed rats suffered diarrhoea, indicated by increased drinking, faeces weight and faecal water content compared with control rats. Urine output was unchanged. With bile acid-feeding, RNA-sequencing revealed 204 increased and 401 decreased mRNAs; mass spectrometry 183 increased and 111 decreased proteins. Among the altered genes were genes associated with electrolyte and water transport (including Slc12a7, Clca4 and Aqp3) and genes associated with bile acid transport (Slc2b1, Abcg2, Slc51a, Slc51b and Fabps). Correlation analysis showed a significant positive correlation (Pearson's r=0.28) between changes in mRNA-expression and changes in protein-expression. However, caution must be exercised in making a direct correlation between experimentally determined transcriptomes and proteomes. Genes associated with bile acid transport responded to bile acid-feeding, suggesting that colonic bile acid transport also occur by regulated protein facilitated mechanisms in addition to passive diffusion. In summary, the study provides annotated rat colonic epithelial cell transcriptome and proteome with response to bile acid-feeding.

Human cecal bile acids: concentration and spectrum

2007 ◽  
Vol 293 (1) ◽  
pp. G256-G263 ◽  
Author(s):  
James P. Hamilton ◽  
Guofeng Xie ◽  
Jean-Pierre Raufman ◽  
Susan Hogan ◽  
Terrance L. Griffin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
Acid Concentration ◽  
Gas Chromatography Mass Spectrometry ◽  
Cecal Content ◽  
Liquid Chromatography Mass Spectrometry ◽  
Bile Acid Concentration ◽  
Chromatography Mass Spectrometry

To obtain information on the concentration and spectrum of bile acids in human cecal content, samples were obtained from 19 persons who had died an unnatural death from causes such as trauma, homicide, suicide, or drug overdose. Bile acid concentration was measured via an enzymatic assay for 3α-hydroxy bile acids; bile acid classes were determined by electrospray ionization mass spectrometry and individual bile acids by gas chromatography mass spectrometry and liquid chromatography mass spectrometry. The 3α-hydroxy bile acid concentration (μmol bile acid/ml cecal content) was 0.4 ± 0.2 mM (mean ± SD); the total 3-hydroxy bile acid concentration was 0.6 ± 0.3 mM. The aqueous concentration of bile acids (supernatant after centrifugation) was identical, indicating that most bile acids were in solution. By liquid chromatography mass spectrometry, bile acids were mostly in unconjugated form (90 ± 9%, mean ± SD); sulfated, nonamidated bile acids were 7 ± 5%, and nonsulfated amidated bile acids (glycine or taurine conjugates) were 3 ± 7%. By gas chromatography mass spectrometry, 10 bile acids were identified: deoxycholic (34 ± 16%), lithocholic (26 ± 10%), and ursodeoxycholic (6 ± 9), as well as their primary bile acid precursors cholic (6 ± 9%) and chenodeoxycholic acid (7 ± 8%). In addition, 3β-hydroxy derivatives of some or all of these bile acids were present and averaged 27 ± 18% of total bile acids, indicating that 3β-hydroxy bile acids are normal constituents of cecal content. In the human cecum, deconjugation and dehydroxylation of bile acids are nearly complete, resulting in most bile acids being in unconjugated form at submicellar and subsecretory concentrations.

scholarly journals Comparison of Various Chromatographic Systems for Analysis of Cytisine in Human Serum, Saliva and Pharmaceutical Formulation by HPLC with Diode Array, Fluorescence or Mass Spectrometry Detection

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2580 ◽  
Author(s):  
Karol Wróblewski ◽  
Anna Petruczynik ◽  
Tomasz Tuzimski ◽  
Dominika Przygodzka ◽  
Grzegorz Buszewicz ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Liquid Chromatography ◽  
Cation Exchange ◽  
Biological Samples ◽  
Stationary Phases ◽  
Reversed Phase ◽  
Pharmaceutical Formulation ◽  
Diode Array ◽  
Strong Cation Exchange ◽  
Pre Treatment

Background: Identification and quantitative determination of cytisine, especially in biological samples and pharmaceutical formulations, is still a difficult analytical task. Cytisine is an alkaloid with a small and very polar molecule. For this reason, it is very weakly retained on reversed phase (RP) stationary phases, such as commonly used alkyl-bonded phases. The very weak retention of cytisine causes it to be eluted together with the components of biological matrices. Objective: Comparison and evaluation of various chromatographic systems for analysis of cytisine in different matrices—serum, saliva and pharmaceutical formulation—by high performance liquid chromatography (HPLC) with diode array (DAD), fluorescence (FLD) and mass spectrometry (MS) detection. Methods: The analyses were performed using HPLC in reversed phase (RP), hydrophilic interaction liquid chromatography (HILIC) and ion exchange chromatography (IEC) modes. Different sample pre-treatment methods were tested: Protein precipitation (with acetone, methanol (MeOH) or acetonitrile (ACN), and solid phase extraction (SPE) using cartridges with octadecyl (C18), hydrophilic-lipophilic balanced copolymer (HLB) or strong cation exchange sorbents (Strata X-C). Conclusion: Significant differences were observed in retention parameters with a change of the used chromatographic system. The various properties of stationary phases resulted in differences in analyte retention, peaks’ shape and systems’ efficiency. The weakest retention was observed using RP systems; however, the use of the Polar RP phase can be an alternative for application in green chromatography. In the strongest retention was observed using a strong cation exchange (SCX) phase. The most optimal systems were chosen for the analysis of cytisine in the pharmaceutical preparation, serum and saliva after sample pre-treatment with the new SPE procedure. Due to the sensitivity, the use of HPLC-DAD or HPLC-FLD is the most optimal for drug analysis in pharmaceutical preparations, whereas HPLC-MS is suitable for analysis of cytisine in biological samples.

scholarly journals Comprehensive Characterization of Bile Acids in Human Biological Samples and Effect of 4-Week Strawberry Intake on Bile Acid Composition in Human Plasma

Metabolites ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 99
Author(s):  
Anqi Zhao ◽  
Liyun Zhang ◽  
Xuhuiqun Zhang ◽  
Indika Edirisinghe ◽  
Britt M. Burton-Freeman ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Human Plasma ◽  
Biological Samples ◽  
Acid Metabolism ◽  
Plasma Concentrations ◽  
Urine Samples ◽  
Bile Acid Metabolism ◽  
Comprehensive Characterization

Primary bile acids (BAs) and their gut microbial metabolites have a role in regulating human health. Comprehensive characterization of BAs species in human biological samples will aid in understanding the interaction between diet, gut microbiota, and bile acid metabolism. Therefore, we developed a qualitative method using ultra-high performance liquid chromatography (UHPLC) coupled with a quadrupole time-of-flight (Q-TOF) to identify BAs in human plasma, feces, and urine samples. A quantitative method was developed using UHPLC coupled with triple quadrupole (QQQ) and applied to a previous clinical trial conducted by our group to understand the bile acid metabolism in overweight/obese middle-aged adults (n = 34) after four weeks strawberry vs. control intervention. The qualitative study tentatively identified a total of 81 BAs in human biological samples. Several BA glucuronide-conjugates were characterized for the first time in human plasma and/or urine samples. The four-week strawberry intervention significantly reduced plasma concentrations of individual secondary BAs, deoxycholic acid, lithocholic acid and their glycine conjugates, as well as glycoursodeoxycholic acid compared to control (p < 0.05); total glucuronide-, total oxidized-, total dehydroxyl-, total secondary, and total plasma BAs were also lowered compared to control (p < 0.05). The reduced secondary BAs concentrations suggest that regular strawberry intake modulates the microbial metabolism of BAs.

scholarly journals Simultaneous analysis of eight benzodiazepines in blood and urine matrix by gas chromatography–mass spectrometry: Implications for air crash investigation

2019 ◽  
Vol 63 ◽  
pp. 2-7
Author(s):  
SR Santosh ◽  
S Sampath ◽  
A Gupta
Keyword(s):  
Mass Spectrometry ◽  
Gas Chromatography ◽  
Biological Samples ◽  
Whole Blood ◽  
Simultaneous Detection ◽  
Gas Chromatography Mass Spectrometry ◽  
Chromatography Mass Spectrometry ◽  
Selective Ion Monitoring ◽  
Urine Matrix ◽  
Detection And Quantification

Introduction: Benzodiazepines are the most commonly prescribed class of drugs in India and are capable of impairing the performance of an aviator in therapeutic to subtherapeutic levels. Detection of benzodiazepines, particularly in blood, is not easy, since the concentrations present, especially following prescribed medical use, can be very low. Several publications have addressed estimation of benzodiazepines in plasma or serum; however, few have attempted their detection in whole blood. Urine, although a better specimen, benzodiazepines due to their extensive metabolism, its metabolites are excreted in urine instead of the parent compounds. Materials and Methods: In our laboratory, a method was developed for simultaneous detection and quantification of eight benzodiazepines in whole blood and urine matrix by gas chromatography–mass spectrometry selective ion monitoring (SIM) method. Chromatographic separation was optimized and achieved for separation of all 8 compounds using Agilent DB-5MS column. Retention time, selectivity and sensitivity were achieved by measuring each analyte in SIM mode. The developed method was tested and validated on actual biological samples for lorazepam, temazepam, diazepam, clonazepam, and nitrazepam. Conclusion: A single method was developed for the detection and quantification of eight benzodiazepines in whole blood and urine matrix by GC–MS SIM method. The method was also tested on limited number of actual biological samples for the lorazepam, temazepam, diazepam, clonazepam, and nitrazepam.

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