High performance liquid chromatographic determination of individual bile acids in serum for automatic diagnosis of various liver and biliary diseases

Zhao, Rui; Li, B. Y.; Chen, N.; Zhang, Y. K.; Wang, Z. Y.; Lu, Ping

doi:10.1002/bmc.1130070307

Cited by 8 publications

(2 citation statements)

References 7 publications

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“…And, efficacy of bile acids determination in various biological samples might be improved by using our proposed methods (activated aminoethyl cellulose membrane bound enzymes & activated polyvinyl alcohol membrane bound enzymes) for clinical diagnosis of liver function dysfunctions, hepatobiliary diseases, adrenocortical diseases and illeal dysfunctions as cost effective alternative. 9 As well as these polymeric membranes bound bile acid enzymes methods can also be considered in future for preparation of bile acid biosensor to carry out more reliable and sensitive analytic method for assay of bile acid in various biological samples.…”

Section: Discussionmentioning

confidence: 99%

Immobilization of bile acid enzymes onto activated amino-ethyl cellulose and polyvinyl alcohol membranes

Rani¹

2020

IJCAP

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Immobilization of bile acid enzymes named, 3-alpha hydroxysteroid dehydrogenase (3-α HSD) and lipoyl dehydrogenase (Diaphorase) enzymes was done on to activated polymeric membranes like amino-ethyl cellulose and Polyvinyl Alcohol. Immobilization of bile acid enzymes was done onto activated aminoethyl cellulose by covalent coupling by using NaOH and glutaraldehyde. And, Immobilization of bile acid enzymes was done onto activated polyvinyl alcohol by using terephthaladehyde and 2-(m-aminophenyl)-1-3-dioxolane. Percent of enzyme coupled was found 65% and 60% for enzymes coupled onto activated amino-ethyl cellulose and activated polyvinyl alcohol respectively. Observed percent of analytical recovery for bile acid estimation was <98% for activated polymeric membrane bound bile acid enzymes. Improved % of retention activities were observed for activated polymeric membranes bound bile acid enzymes in our present method even after the regular use of more than 4 months (16 weeks) when preserved at 4 dgree C. Hence, our present method of immobilization of bile acid enzymes onto activated polymeric membranes were found to be more cost effective as compared to other reported enzyme based methods.

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Section: Discussionmentioning

confidence: 99%

Immobilization of bile acid enzymes onto activated amino-ethyl cellulose and polyvinyl alcohol membranes

Rani¹

2020

IJCAP

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“…Therefore, the separation and quantification of BA are very important for the diagnosis, prognosis and clarification of pathophysiology, but present difficulties because of their low serum levels. High-performance liquid chromatography (HPLC) is well recognized as an effective tool for the analysis of unconjugated and conjugated BA (Coca et al, 1994;Goldsmith et al, 1994;Goto, 1990;Nambara and Goto, 1988;Nichols et al, 1993;Zhang et al, 1992;Zhao et al, 1993). These compounds, however, do not present significant UV absorption, fluorescent or electrochemical properties suitable for their sensitive detection.…”

Section: Introductionmentioning

confidence: 97%

HPLC-Fluorescence Determination of Individual Free and Conjugated Bile Acids in Human Serum

Gatti

Roda

Cerrè

et al. 1997

Biomed. Chromatogr.

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A method for the quantitative analysis of unconjugated and conjugated bile acids (BA) in serum of patients with primary biliary cirrhosis (PBC) before and after therapy with antibiotic or ursodeoxycholic acid (UDCA) is described. After separation of the free, glycine and taurine conjugated (F, G and T conjugated) fractions by solid‐phase extraction, the isolated T conjugates were hydrolysed enzymatically using cholyglycine hydrolase. The BA fractions were derivatized using 2‐bromoacetyl‐6‐methoxynaphthalene (Br‐AMN) and detected fluorimetrically (λexc=300 nm, λem=460 nm). The derivatization reaction was performed under mild conditions (10 min at 40°C) in an aqueous medium in the presence of tetrakis (decyl) ammonium bromide (TDeABr). The HPLC separation was achieved using an ODS column and with a mobile phase gradient mixture of A–B, where A is water and B is acetonitrile:methanol (60:40 v/v) for elution at a flow‐rate of 1.2 mL/min. The reproducibility, recovery and separation of individual BA under gradient elution conditions were satisfactory, allowing a sensitive detection of each BA in serum samples with a detection limit of about 1–2 pmol. © 1997 by John Wiley & Sons, Ltd.

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Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

You¹,

Shi

Zhao

et al. 2006

J. Sep. Sci.

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A highly sensitive and accurate method based on the precolumn derivatization of bile acids (BA) with a high ionization efficiency labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-benzenesulfonate (BDEBS) coupled with LC/MS has been developed. After derivatization, BA molecules introduced a weak basic nitrogen atom into the molecular core structure that was readily ionized in commonly used acidic HPLC mobile phases. Derivatives were sufficiently stable to be efficiently analyzed by atmospheric pressure chemical ionization (APCI)-MS/MS in positive-ion mode. The MS/MS spectra of BA derivatives showed an intense protonated molecular ion at m/z [M + H]+. The collision-induced dissociation of the molecular ion produced fragment ions at [MH-H2O]+, [MH-2H2O]+, [MH-3H2O]+. The characteristic fragment ions were at m/z 320.8, 262.8, and 243.7 corresponding to a cleavage of N-CO, O-CO, and C-OCO, respectively, and bonds of derivatized molecules. The selected reaction monitoring, based on the m/z [M+H]+ --> [MH-H2O]+, [MH-2H2O]+, [MH-3H2O]+, 320.8, 262.8, and 243.7 transitions, was highly specific for the BA derivatives. The LODs for APCI in a positive-ion mode, at an S/N of 5, were 44.36-153.6 fmol. The validation results showed high accuracy in the range of 93-107% and the mean interday precision for all standards was <15% at broad linear dynamic ranges (0.0244-25 nmol/mL). Good linear responses were observed with coefficients of > 0.9935 in APCI/MS detection. Therefore, the facile BDEBS derivatization coupled with mass spectrometric analysis allowed the development of a highly sensitive and specific method for the quantitation of trace levels of the free and glycine-conjugated BA from human serum samples.

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High performance liquid chromatographic determination of individual bile acids in serum for automatic diagnosis of various liver and biliary diseases

Cited by 8 publications

References 7 publications

Immobilization of bile acid enzymes onto activated amino-ethyl cellulose and polyvinyl alcohol membranes

Immobilization of bile acid enzymes onto activated amino-ethyl cellulose and polyvinyl alcohol membranes

HPLC-Fluorescence Determination of Individual Free and Conjugated Bile Acids in Human Serum

Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

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