Significance of Morphine Concentration in Bile, Liver, and Blood

Abstract

Forensic pathologists are requested to select matrices alternative to blood in cases of toxicological interest in which blood is not available for different reasons. We evaluated morphine concentrations in blood, bile, and liver samples in 52 cases of heroin overdoses, relating them to each other, to understand the information that could be derived from their analysis. Gas chromatography/mass spectrometry analysis was performed for all the samples positive on screening for opiates. Shapiro-Wilk test, nonparametric Mann-Whitney test, linear regression analysis, and Bland-Altman test were used for analysis. Linear regression demonstrated that there was not a statistically significant association in morphine concentrations between blood and bile and blood and liver. Mean liver/blood ratio was 2.76, varying from 0.131 to 13.379, and bile/blood ratio was 28.79, varying from 0.28 to 559.16. According to these results, bile analysis is a “screening test”; biliary or hepatic concentration of morphine cannot provide information on hematic concentration at the time of death, having no forensic value taken individually.

Heroin-Related Fatalities in Jeddah, Saudi Arabia, between 2008 and 2018

“…Many factors that affect the connections between heroin biomarkers and their metabolites must be taken into consideration, such as the history of abuse, concomitant use of other drugs, tolerance, route of administration, and PMI [ 3 , 33 , 44 ]. Mercurio et al [ 98 ] studied 52 morphine-related deaths in which blood and bile were assessed in parallel and found that bile/blood morphine concentrations were poorly correlated ( r 2 = 0.01, p = 0.4), which agrees with the current investigation. In that study, the median ratio of free morphine between the bile and blood was twenty-nine, which was lower than that in the current study.…”

Postmortem Liver and Kidney Tissue Concentrations of Heroin Biomarkers and Their Metabolites in Heroin‐Related Fatalities*†

“…Since then, multiple studies have reported morphine concentrations in liver and blood simultaneously with a wide range of concentrations reported, and morphine concentration liver to blood ratios ranging from 0.4 to 2.4 (16,22,29,30). Previous studies reported median total morphine concentrations in blood of 110.0 ng/mL (range: 10.0-570.0 ng/mL) (22), 530 ng/mL (range: 20.0-3540.0 ng/mL) (23), and 600 ng/mL (range: 70.0-1600.0 ng/mL) (15). Liver has higher values with medians between 320.0 and 1270.0 ng/g, minimums of 40 ng/g, and maximums of 5728 ng/g (15,23) To provide greater insight, Spiehler and Brown (20) subdivided their cases into rapid (56 cases) or delayed deaths (145 cases).…”

“…Previous studies reported median total morphine concentrations in blood of 110.0 ng/mL (range: 10.0–570.0 ng/mL) (22), 530 ng/mL (range: 20.0–3540.0 ng/mL) (23), and 600 ng/mL (range: 70.0–1600.0 ng/mL) (15). Liver has higher values with medians between 320.0 and 1270.0 ng/g, minimums of 40 ng/g, and maximums of 5728 ng/g (15,23) To provide greater insight, Spiehler and Brown (20) subdivided their cases into rapid (56 cases) or delayed deaths (145 cases). In rapid deaths, the median morphine concentrations were higher than in delayed deaths, and in contrast, in delayed deaths, morphine concentrations in liver were higher than in rapid deaths.…”

“…In addition, their extraction recovery was better than 70.0%. Other reported methods have measured total morphine after converting glucuronide conjugates of morphine to their free form with enzymatic hydrolysis (15,23) or have used direct determination of morphine and its glucuronide conjugates without hydrolysis using LC-MS-MS (24). In fact, few studies have reported the level of heroin metabolites in solid tissue specimens.…”

“…Most of the procedures detailed above used GC. Most of the important metabolites are found in their conjugated form, which requires hydrolysis and derivatization procedures to prepare samples for GC-MS, which is time consuming, tedious, and dangerous (5,15,16,19,20,22,23,25).…”