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 determ ination of morphine and its glucuronides in human blood is described. For t he preparation of an immunoadsorber, specific antisera (polyclonal, host: r abbit) against morphine, morphine-3-glucuronide and morphine-6-glucuronide were coupled to 1,1'-carbonyldiimidazole-activated tris-acrylgel and used f or immunoaffinity extraction of morphine and its glucuronides from coronary blood. The resulting extracts were analysed by HPLC with native fluorescen ce detection. The mean recoveries from spiked blood samples were 71%, 76% a nd 88% for morphine, morpkine-3-glucuronide and morphine-6-glucuronide, res pectively. The Limit of detection was 3 ng/g blood and the limit of quantit ation 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. (C) 1999 Elsevier Science B.V. All rights reserved.