Dataset Information


Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry.

ABSTRACT: Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization⁻mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, using bacterial cholesterol oxidase enzyme to convert the 3β-hydroxy group to a 3-oxo group for subsequent derivatization with the positively charged Girard hydrazine reagents, or on substrates with a native oxo group. Here we describe an extension of the technology by substituting 3α-hydroxysteroid dehydrogenase (3α-HSD) for cholesterol oxidase, making the method applicable to sterols with a 3α-hydroxy-5β-hydrogen structure. The 3α-HSD enzyme works efficiently on bile alcohols and bile acids with this stereochemistry. However, as found by others, derivatization of the resultant 3-oxo group with a hydrazine reagent does not go to completion in the absence of a conjugating double bond in the sterol structure. Nevertheless, Girard P derivatives of bile alcohols and C27 acids give an intense molecular ion ([M]⁺) upon electrospray ionization and informative fragmentation spectra. The method shows promise for analysis of bile alcohols and 3α-hydroxy-5β-C27-acids, enhancing the range of sterols that can be analyzed at high sensitivity in sterolomic studies.

SUBMITTER: Abdel-Khalik J 

PROVIDER: S-EPMC6384595 | BioStudies | 2019-01-01

REPOSITORIES: biostudies

Similar Datasets

2012-01-01 | S-EPMC3351325 | BioStudies
2016-01-01 | S-EPMC4916447 | BioStudies
2017-01-01 | S-EPMC5649119 | BioStudies
2019-01-01 | S-EPMC6651411 | BioStudies
2019-01-01 | S-EPMC6327153 | BioStudies
2016-01-01 | S-EPMC5089630 | BioStudies
2019-11-05 | GSE130336 | GEO
2020-01-01 | S-EPMC7084107 | BioStudies
1980-01-01 | S-EPMC1162388 | BioStudies
2015-06-30 | E-GEOD-64976 | ArrayExpress