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Molecules 2014, 19(1), 1189-1200; doi:10.3390/molecules19011189
Article

Validation of a Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Method for Determination of All-Trans Retinoic Acid in Human Plasma and Its Application to a Bioequivalence Study

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Received: 29 November 2013; in revised form: 25 December 2013 / Accepted: 7 January 2014 / Published: 17 January 2014
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Abstract: A sensitive, reliable and specific LC-MS-MS method was developed and validated for the identification and quantitation of all-trans retinoic acid (ATRA) in human plasma. Acitretin was used as the internal standard (IS). After liquid-liquid extraction of 500 μL plasma with methyl tert-butyl ether (MTBE), ATRA and the IS were chromatographed on a HyPURITY C18 column (150 mm × 2.1 mm, 5 μm) with the column temperature set at 40 °C. The mobile phase was consisted of 40% phase A (MTBE–methanol–acetic acid, 50:50:0.5, v/v) and 60% phase B (water–methanol–acetic acid, 50:50:0.5, v/v) with a flow rate of 0.3 mL/min. The API 4000 triple quadrupole mass spectrometer was operated in multiple reaction monitoring (MRM) mode via the positive electrospray ionization interface using the transition m/z 301.4 → 123.1 for ATRA and m/z 326.9 → 177.1 for IS, respectively. The calibration curve was linear over the range of 0.45–217.00 ng/mL (r ≥ 0.999) with a lower limit of quantitation (LLOQ) of 0.45 ng/mL. The intra- and inter-day precisions values were below 8% relative standard deviation and the accuracy was from 98.98% to 106.19% in terms of relative error. The validated method was successfully applied in a bioequivalence study of ATRA in Chinese healthy volunteers.
Keywords: all-trans retinoic acid (ATRA); LC-MS-MS; liquid-liquid extraction; bioequivalence all-trans retinoic acid (ATRA); LC-MS-MS; liquid-liquid extraction; bioequivalence
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

Peng, J.-B.; Luo, C.-H.; Wang, Y.-C.; Huang, W.-H.; Chen, Y.; Zhou, H.-H.; Tan, Z.-R. Validation of a Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Method for Determination of All-Trans Retinoic Acid in Human Plasma and Its Application to a Bioequivalence Study. Molecules 2014, 19, 1189-1200.

AMA Style

Peng J-B, Luo C-H, Wang Y-C, Huang W-H, Chen Y, Zhou H-H, Tan Z-R. Validation of a Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Method for Determination of All-Trans Retinoic Acid in Human Plasma and Its Application to a Bioequivalence Study. Molecules. 2014; 19(1):1189-1200.

Chicago/Turabian Style

Peng, Jing-Bo; Luo, Chen-Hui; Wang, Yi-Cheng; Huang, Wei-Hua; Chen, Yao; Zhou, Hong-Hao; Tan, Zhi-Rong. 2014. "Validation of a Liquid Chromatography-Electrospray Ionization-Tandem Mass Spectrometry Method for Determination of All-Trans Retinoic Acid in Human Plasma and Its Application to a Bioequivalence Study." Molecules 19, no. 1: 1189-1200.



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