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A rapid and sensitive bioassay based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed and validated for the simultaneous determination of codeine and its active metabolites, including morphine, morphine 3β-glucuronide (M3G) and morphine 6β-glucuronide (M6G), in human plasma. Sample preparation of plasma after the addition of naloxone as internal standard (IS) involved solid-phase extraction (SPE) on C18 cartridges. Reversed-phase chromatography using a gradient elution with methanol and 0.04% formic acid solution (pH 3.5) was used for separation in a run time of 5 min. The analytes were detected in the positive ion mode using multiple reaction monitoring (MRM) of the transitions at m/z 300.4→215.2 for codeine, 286.2→152.0 for morphine, and 462.2→286.2 for M3G and M6G. The method has the following performance characteristics: a reliable response range of 0.05-80 ng/ml for codeine, M3G and M6G and a response range of 0.05-5.0 ng/ml for morphine with correlation coefficients (r) of >0.997 for all analytes. The lower limit of quantitation (LLOQ) for all four analytes was 0.05 ng/ml. The intra- and inter-day precision and accuracy of the quality control samples at low, medium and high concentration levels showed <12% relative standard deviation (RSD) and -6.9 to 8.1% relative error (RE) for all the analytes. The method was successfully applied to a pharmacokinetic study of codeine in healthy Mongolian Chinese volunteers after a 30 mg oral dose. Copyright © 2013 Elsevier B.V. All rights reserved.

Citation

Xiujun Wu, Weiping Zhang, Yin Bai, Tao Guo, Jingkai Gu. Simultaneous analysis of codeine and its active metabolites in human plasma using liquid chromatography-tandem mass spectrometry: application to a pharmacokinetic study after oral administration of codeine. Journal of pharmaceutical and biomedical analysis. 2013 May 5;78-79:261-8

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PMID: 23507688

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