TITLE

Determination of naltrexone and 6β-naltrexol in human blood: comparison of high-performance liquid chromatography with spectrophotometric and tandem-mass-spectrometric detection

AUTHOR(S)
Brünen, Sonja; Krüger, Ralf; Finger, Susann; Korf, Felix; Kiefer, Falk; Wiedemann, Klaus; Lackner, Karl J.; Hiemke, Christoph
PUB. DATE
February 2010
SOURCE
Analytical & Bioanalytical Chemistry;Feb2010, Vol. 396 Issue 3, p1249
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
We present data for a comparison of a liquid-chromatographic method coupled with tandem mass spectrometry (LC-MS/MS) and a high-performance liquid-chromatographic method with column switching and UV spectrophotometric detection. The two methods were developed for determination of naltrexone and 6β-naltrexol in blood serum or plasma aiming to be used for therapeutic drug monitoring to guide the treatment of patients with naltrexone. For the high-performance liquid chromatography (HPLC)/UV detection, online sample cleanup was conducted on Perfect Bond C18 material with 2% (vol/vol) acetonitrile in deionized water. Drugs were separated on a C18 column using 11.5% (vol/vol) acetonitrile and 0.4% (vol/vol) N, N, N, N-tetramethylethylenediamine within 20 min. LC-MS/MS used naltrexone- d3 and 6β-naltrexol- d4 as internal standards. After protein precipitation, the chromatographic separation was performed on a C18 column by applying a methanol gradient (5–100%, vol/vol) with 0.1% formic acid over 9.5 min. The HPLC/UV method was found to be linear for concentrations ranging from 2 to 100 ng/ml, with a regression correlation coefficient of r2 > 0.998 for naltrexone and 6β-naltrexol. The limit of quantification was 2 ng/ml for naltrexone and 6β-naltrexol. For the LC-MS/MS method the calibration curves were linear ( r² > 0.999) from 0.5 to 200 ng/ml for both substances, and the limit of quantification was 0.5 ng/ml. The concentrations measured by the two methods correlated significantly for both substances ( r² > 0.967; p < 0.001). Both methods could be used for therapeutic drug monitoring. The HPLC/UV method was advantageous regarding automatization and costs, whereas LC-MS/MS was superior with regard to sensitivity.
ACCESSION #
47656752

 

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