國家衛生研究院 NHRI:Item 3990099045/3014

NHRI > National Institute of Cancer Research > Jacqueline Whang-Peng(1996-2007) > Periodical Articles > Item 3990099045/3014

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Title:	Novel inhibition of cis/trans retinoic acid interconversion in biological fluids - an accurate method for determination of trans and 13-cis retinoic acid in biological fluids
Authors:	Wang, CJ;Pao, LH;Hsiong, CH;Wu, CY;Whang-Peng, JJK;Hu, OYP
Contributors:	National Institute of Cancer Research
Abstract:	All-trans retinoic acid (tRA, or tretinoin) can be metabolized through stereoisomerzation to 13-cis retinoic acid (13-cRA) in vivo. We have developed a simple, sensitive and accurate method for analyzing tRA and 13-cRA in plasma with the addition of N-ethylmaleimide (NEM) and Vitamin C (Vit. C) to prevent the interconversion of cis/trans retinoic acid. All-trans RA, 9-cRA, and 13-cRA were well separated from each other in plasma by using a C18 precolumn and a column with a gradient solvent system of mobile phases A and B at a flow rate of 1.0 ml/min. In addition, thermal stability of tRA and cRA in plasma during the sample preparation under the temperature of 0 and 25 degreesC were studied. Our results showed that (1) the interconversion ratios (%) (cRA/tRA and tRA/cRA) were decreased with the addition of NEM and Vit. C and the minimum concentrations of NEM and Vit. C to inhibit the interconversion were 50 and 150 muM, respectively, (2) higher concentrations of NEM and Vit. C were required to prevent the interconversion at higher temperature, (3) the precision and accuracy of calibration curve with various concentration of tRA (1-1000 ng/ml) and 13-cRA (5-800 ng/ml) in plasma showed good linearity (r(2) = 0.9992 and 0.9994), and between-day errors expressed by coefficient of variation (CV, %) for tRA and 13-cRA which were both less than 5.6%, (4) the mean recovery of the analytes were 78-94% for tRA and 80-92% for 13-cRA at concentration range from 1 to 1000 ng/ml and 5 to 800 ng/ml, respectively, and (5) the limit of quantitation of tRA and 13-cRA were 1 and 5 ng/ml, respectively. In addition, the HPLC method had been successfully applied to the tRA pharmacokinetic study in two hepatoma patients after receiving 45 mg/m(2) per day orally. Thus, our results suggest that the HPLC method for analyzing tRA and 13-cRA in plasma with the addition of NEM and Vit. C is a simple, sensitive and accurate method.
Keywords:	Biochemical Research Methods;Chemistry, Analytical
Date:	2003-11-05
Relation:	Journal of Chromatography B. 2003 Nov;796(2):283-291.
Link to:	http://dx.doi.org/10.1016/S1570-0232(03)00572-5
JIF/Ranking 2023:	http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=1570-0232&DestApp=IC2JCR
Cited Times(WOS):	https://www.webofscience.com/wos/woscc/full-record/WOS:000186381300007
Cited Times(Scopus):	http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0142227110
Appears in Collections:	[Jacqueline Whang-Peng(1996-2007)] Periodical Articles

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