scholarly journals An Integrated Metabolomic Study of Osteoporosis: Discovery and Quantification of Hyocholic Acids as Candidate Markers

2021 ◽  
Vol 12 ◽  
Author(s):  
Dawei Deng ◽  
Chen Pan ◽  
Zeming Wu ◽  
Yujiao Sun ◽  
Chang Liu ◽  
...  
Keyword(s):  
Bile Acids ◽  
High Performance ◽  
High Resolution Mass Spectrometry ◽  
Serum Samples ◽  
Treatment Of Osteoporosis ◽  
The Family ◽  
Prevalent Disease ◽  
Gene Correlation ◽  
Prevention Of Osteoporosis ◽  
Lipids Metabolism

Osteoporosis is becoming a highly prevalent disease in a large proportion of the global aged population. Serum metabolite markers may be important for the treatment and early prevention of osteoporosis. Serum samples from 32 osteoporosis and 32 controls were analyzed by untargeted metabolomics and lipidomic approaches performed on an ultra-high performance liquid chromatography and high-resolution mass spectrometry (UHPLC-HRMS) system. To find systemic disturbance of osteoporosis, weighted gene correlation network analysis (WGCNA) and statistical methods were employed for data-mining. Then, an in-depth targeted method was utilized to determine potential markers from the family of key metabolites. As a result, 1,241 metabolites were identified from untargeted methods and WGCNA indicated that lipids metabolism is deregulated and glycerol phospholipids, sphingolipids, fatty acids, and bile acids (BA) are majorly affected. As key metabolites of lipids metabolism, 66 bile acids were scanned and 49 compounds were quantified by a targeted method. Interestingly, hyocholic acids (HCA) were found to play essential roles during the occurrence of osteoporosis and may be potential markers. These metabolites may be new therapeutic or diagnosis targets for the screening or treatment of osteoporosis. Quantified measurement of potential markers also enables the establishment of diagnostic models for the following translational research in the clinic.

scholarly journals Methods for the Preparation of Blood Serum Samples in the Assessment of the Pharmacokinetics of Protein Drugs by the Example of Viscumin

2021 ◽  
Vol 4 (3) ◽  
pp. e00152
Author(s):  
N.S. Yudina ◽  
Ya.N. Barashkova ◽  
O.V. Vladimirova ◽  
V.A. Myasnikov
Keyword(s):  
Blood Serum ◽  
High Performance ◽  
Serum Proteins ◽  
High Resolution Mass Spectrometry ◽  
Viscum Album ◽  
Serum Samples ◽  
Extraction Degree ◽  
Degree Of Extraction ◽  
Alcohol Testing

A comparative evaluation of methods for preparing blood serum samples for determination of the lectin content of the mistletoe Viscum Album, viscumin, by high-performance liquid chromatography in combination with high-resolution mass-spectrometry detection has been carried out. Based on the results of the analysis of the lectin hydrolyzate, a specific peptide fragment with m/z 791.388 suitable for subsequent quantitative determination was selected. Isolation of viscumin from serum components was carried out without the use of specific antibodies. The study used methods employing various spin columns and the method of protein precipitation with 1% TCA (trichloroacetic acid) in IPA (isopropyl alcohol). Testing the methods of depletion of blood serum proteins was based on the determination of the degree of extraction of lectin from model serum samples. As a result of our research, we have developed a technique for the quantitative analysis of viscumin in blood serum by the most efficient method of sample preparation using the ProteoMiner spin columns. The technique allows to determine the protein concentration up to 5·10-4 mg/ml with the extraction degree (2.7 ± 0.6) %.

Steroidomics for highlighting novel serum biomarkers of testosterone doping

Bioanalysis ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 1169-1185 ◽  
Author(s):  
Federico Ponzetto ◽  
Julien Boccard ◽  
Raul Nicoli ◽  
Tiia Kuuranne ◽  
Martial Saugy ◽  
...  
Keyword(s):  
High Performance ◽  
Biomarker Discovery ◽  
High Resolution Mass Spectrometry ◽  
Serum Markers ◽  
Serum Biomarkers ◽  
Serum Samples ◽  
Steroid Profiling ◽  
Efficient Alternative ◽  
Detection Capabilities ◽  
Endogenous Steroid

Aim: Quantification of testosterone (T) and 5α-dihydrotestosterone serum concentrations proved to be an efficient alternative to urinary steroid profiling for the detection of T doping. In this context, additional serum markers could be discovered by exploratory untargeted steroidomics studies. Results: Endogenous steroid metabolites were monitored by ultra high-performance liquid chromatography coupled to high-resolution mass spectrometry in serum samples collected during a T administration clinical trial. A three-step workflow for accurate review of annotation was used and multifactorial data analysis allowed highlighting promising serum biomarkers. Longitudinal monitoring of selected compounds was performed to assess T abuse detection capabilities. Conclusion: Application of serum steroidomics showed high potential for biomarker discovery of T doping, suggesting longitudinal monitoring of steroid hormones in serum as a significant improvement in detection of endogenous steroids abuse.

scholarly journals Determination of Conjugated and Unconjugated Serum 3α-OH Bile Acids by High-Performance Liquid Chromatography

1994 ◽  
Vol 31 (5) ◽  
pp. 479-484 ◽  
Author(s):  
R F Goldsmith ◽  
M Gruca ◽  
M T O'Halloran ◽  
J W Earl ◽  
K Gaskin
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Bile Acid ◽  
Bile Acids ◽  
High Performance ◽  
Clinical Laboratory ◽  
Limit Of Detection ◽  
Hplc Method ◽  
Serum Samples ◽  
Liquid Chromatographic

A simple reverse-phase liquid chromatographic method has been developed for the quantitative measurement of 11 3α-OH bile acids in paediatric serum samples. Bile acids are enzymatically reduced to the corresponding 3-keto compound and then derivatized with the fluorophore dansyl hydrazine. Preliminary fractionation of bile acids is not required. The limit of detection is 0·1μmol/L using a sample size of 200μL. One hundred and twenty-three serum samples were analysed by the high-performance liquid chromatography (HPLC) method and the results compared with a commercial kit method (Enzabile) presently used in many laboratories for the measurement of total bile acids. There was a good correlation between the two methods ( r = 0·96). The method is suitable for use in a clinical laboratory for the further investigation of those patients with abnormally high total bile acid levels where quantification of bile acid fractions is required.

Reference ranges of serum bile acids in children and adolescents

2015 ◽  
Vol 53 (11) ◽  
Author(s):  
Jörg Jahnel ◽  
Evelyn Zöhrer ◽  
Hubert Scharnagl ◽  
Wolfgang Erwa ◽  
Günter Fauler ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Children And Adolescents ◽  
Bile Acids ◽  
High Performance ◽  
High Resolution Mass Spectrometry ◽  
Age Groups ◽  
Reference Ranges ◽  
Healthy Children ◽  
Age Dependent ◽  
Resolution Mass

AbstractBile acids (BA) are found predominantly in bile but also in serum, where they can be used as markers for inborn and acquired hepatobiliary disorders. We measured serum BA levels by mass spectrometry to determine reference ranges for healthy children and adolescents in different age groups.In 194 healthy children and adolescents (0–19 years) concentrations of serum BA and BA composition were determined using high-performance liquid chromatography high-resolution mass spectrometry. Individuals were classified by ages into five groups: 0–5 months, 6–24 months, 3–5 years, 6–11 years, and >11 years.The 95% confidence interval of serum total BA values in newborns was 3.85–6.32 μmol/L. In the cohort aged 6–24 months total BA values were significantly higher (6.61–9.43 μmol/L; p<0.001). During growth, values decreased (6–11 years; 3.61–5.41 μmol/L), and after 11 years (3.09–4.12 μmol/L) resembled those in adults (0.28–6.50 μmol/L). With respect to conjugation patterns, in neonates BA were primarily conjugated with taurine; however, after 6 months glycine conjugates clearly predominated.: Our data show that serum BA values vary substantially during the first years of life and that reference ranges for BA are age-dependent. The physiologic mechanisms underlying these variations remain to be determined.

Continuous-flow determination of primary bile acids, by bioluminescence, with use of nylon-immobilized bacterial enzymes.

1984 ◽  
Vol 30 (2) ◽  
pp. 206-210 ◽  
Author(s):  
A Roda ◽  
S Girotti ◽  
S Ghini ◽  
B Grigolo ◽  
G Carrea ◽  
...  
Keyword(s):  
Bile Acid ◽  
Bile Acids ◽  
Continuous Flow ◽  
High Performance ◽  
Immobilized Enzymes ◽  
Serum Samples ◽  
Light Emitting ◽  
Assay Tube ◽  
Bioluminescence Method

Abstract We describe a continuous-flow bioluminescence method for measuring primary bile acid in serum, with nylon as the solid support. 7 alpha-Hydroxysteroid dehydrogenase (EC 1.1.1. 159), a bacterial luciferase (EC 1.14.14.3), and NAD+:FMN oxido-reductase (EC 1.6.8.1) are covalently co-immobilized on a nylon coil (1 m X 1.0 mm i.d.). The assay is highly specific for 7 alpha-hydroxy bile acids. Other bile acids and steroids do not interfere. The continuous-flow light-emitting system, in which the reactor (nylon coil) is placed in front of a photomultiplier tube inside a luminometer, is versatile and simple. The flow is air-segmented, and serum samples (5-50 microL) can be injected directly. Concentration and response are linearly related from 10 to 2500 pmol per assay tube. The precision of the method is satisfactory (CV 5-10%), both inter- and intra-assay. We validated the technique by comparing results with those by RIA, enzyme immunoassay, and "high-performance" liquid chromatography. More than 20 samples an hour can be analyzed, with no carryover. The nylon-immobilized enzymes are stable for more than two months, and greater than 500 samples can be analyzed with use of a few milligrams of enzymes. Normal values for bile acid content of serum ranged from 1 to 2.5 mumol/L, in agreement with those obtained by other methods.

scholarly journals Conjugated Bile Acids in Gallbladder Bile and Serum as Potential Biomarkers for Cholesterol Polyps and Adenomatous Polyps

2016 ◽  
Vol 31 (1) ◽  
pp. 73-79 ◽  
Author(s):  
Mei-Fen Zhao ◽  
Peng Huang ◽  
Chun-Lin Ge ◽  
Tao Sun ◽  
Zhi-Gang Ma ◽  
...  
Keyword(s):  
Bile Acids ◽  
High Performance ◽  
Gallbladder Bile ◽  
Adenomatous Polyps ◽  
Control Group ◽  
Serum Samples ◽  
Diagnostic Efficacy ◽  
Glycocholic Acid ◽  
Bile Samples ◽  
Conjugated Bile Acids

Purpose To identify conjugated bile acids in gallbladder bile and serum as possible biomarkers for cholesterol polyps (CPs) and adenomatous polyps (APs). Methods Gallbladder bile samples and serum samples were collected from 18 patients with CPs (CP group), 9 patients with APs (AP group), and 20 patients with gallstones (control group) from March to November, 2013. High performance liquid chromatography (HPLC) assay with ultraviolent detection was used to detect the concentration of 8 conjugated bile acids (glycocholic acid, GCA; taurocholic acid, TCA; glycochenodeoxycholic acid, GCDCA; taurochenodeoxycholic acid, TCDCA; glycodeoxycholic acid, GDCA; taurodeoxycholic acid, TDCA; taurolithocholic acid, TLCA; tauroursodeoxycholic acid, TUDCA) in bile samples and serum samples. The diagnostic efficacy of serum GCA, GCDCA and TCDCA was evaluated. Results These 8 conjugated bile acids in gallbladder bile and serum were completely identified within 10 minutes with good linearity (correlation coefficient: R>0.9900; linearity range: 3.91-500 µg/mL). Among these conjugated bile acids, the levels of gallbladder bile GCDCA and TCDCA in the CP group were significantly higher than those in the AP group (p<0.05). Furthermore, serum GCDCA and TCDCA as well as GCA were significantly higher in the AP group than the CP group (p<0.05). Serum GCDCA alone (≤12 µg/mL) had relatively better diagnostic efficacy than the other conjugated bile acids. Conclusions The levels of serum GCA, GCDCA and TCDCA may be valuable for differentiation of APs and CPs.

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