Immunoaffinity extraction of morphine, morphine-3-glucuronide and morphine-6-glucuronide from blood of heroin victims for simultaneous high-performance liquid chromatographic determination.
The development of an immunoaffinity-based extraction method for the determination of morphine and its glucuronides in human blood is described. For the preparation of an immunoadsorber, specific antisera (polyclonal, host: rabbit) against morphine, morphine-3-glucuronide and morphine-6-glucuronide were coupled to 1,1'-carbonyldiimidazole-activated tris-acrylgel and used for immunoaffinity extraction of morphine and its glucuronides from coronary blood. The resulting extracts were analysed by HPLC with native fluorescence detection. The mean recoveries from spiked blood samples were 71%, 76% and 88% for morphine, morphine-3-glucuronide and morphine-6-glucuronide, respectively. The limit of detection was 3 ng/g blood and the limit of quantitation was 10 ng/g blood for all three analytes. The results of the analysis of coronary blood samples from 23 fatalities due to heroin are presented.
Beike
J
J
Institute of Legal Medicine, Westfälische Wilhelms-Universität, Münster, Germany.
Köhler
H
H
Brinkmann
B
B
Blaschke
G
G
D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D062787:Drug Overdose / Q000097:blood / Q000401:mortality; D003932:Heroin / Q000506:poisoning*; D006801:Humans; D009020:Morphine / Q000097:blood*; D009022:Morphine Derivatives / Q000097:blood*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D009022:Morphine Derivatives; C035349:morphine-6-glucuronide; D003932:Heroin; D009020:Morphine; C018108:morphine-3-glucuronide