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Development of an LC-MS/MS method for aromatase inhibitor screening

Authors
Park, Myeong HyeonKim, In SookDong, Mi-SookYoo, Hye Hyun
Issue Date
5월-2014
Publisher
SPRINGER HEIDELBERG
Keywords
Aromatase; CYP 19A1; Inhibition; LC-MS/MS
Citation
ANALYTICAL AND BIOANALYTICAL CHEMISTRY, v.406, no.14, pp.3443 - 3449
Indexed
SCIE
SCOPUS
Journal Title
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume
406
Number
14
Start Page
3443
End Page
3449
URI
https://scholar.korea.ac.kr/handle/2021.sw.korea/98697
DOI
10.1007/s00216-014-7764-1
ISSN
1618-2642
Abstract
Aromatase (CYP 19A1) is a key steroidogenic enzyme that catalyzes the conversion of androgen to estrogen. In this study, a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for aromatase inhibitor screening was developed and validated. The substrate androstenedione was incubated with human CYP 19A1 supersomes in the presence of NADPH for 30 min, and estrone formation was determined by LC-MS/MS analysis. Cortisone was used as internal standard. The incubation mixture was extracted using a liquid-liquid extraction method with ethyl acetate. Chromatographic separation was achieved using a C-18 column (3.0 x 50 mm, 2.7 mu m) with a mobile phase consisting of 0.1 % formic acid/acetonitrile adopting gradient elution at a flow rate of 0.4 mL/min. The mass spectrometer was operated in positive electrospray ionization mode. The precursor-product ion pairs used for multiple reaction monitoring were m/z 287 -> 97 (androstenedione), m/z 271 -> aEuro parts per thousand 159 (estrone), and m/z 361 -> aEuro parts per thousand 163 (IS, cortisone). The developed method met the required criteria for the validation of bioanalytical methods. The validated method was successfully applied to evaluate aromatase inhibitory activity of plants extracts of Simaroubaceae.
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