Kamal A. Hadidi

Current trends in drug abuse associated fatalities - Jordan, 2000-2004

This study is the first study that addresses drug abuse associated fatalities in Jordan. It is aimed to give a close picture to the demography, toxicological data, manner, cause of death and other associated findings in such cases. Postmortem forensic pathology reports for all autopsies examined in the National Institute of Forensic Medicine were reviewed over a 5-year period and drug abuse associated deaths were selected. The study revealed that 44 cases (0.76%) out of the 5789 total autopsies were attributed to drug abuse associated deaths. The age range was from 20 to 60 years (mean+/-S.D.=32.7+/-7.2). More than 80% of cases were Jordanian males. The reported abuse substances as single drug or in combination were alcohol in 56.8%, morphine 36.4%, heroin 15.9%, benzodiazepines in 11.4% and cocaine in one body packer case. Surprisingly, neither a case with amphetamine or amphetamine analogue, nor with marijuana or methadone was recorded. In 75% of cases the death was accidental and only one case was reported to be suicidal, while in 18.2% and 4.5% death was due to sudden death and road traffic accidents, respectively. Regarding the cause of death, it was related to drug overdose in 50% of cases and in 34.1%, 11.4% and 4.5% of cases it was attributed to drug related medical complications, non-drug related complications, and trauma, respectively. Alcohol was mainly associated with accidental death; morphine and heroin were associated with drug overdose and abused through intravenous route. Injection marks were reported in 56.8% of cases and in 52.3% death occurred at home. This study confirmed the variation in the incidence and type of abused substances in Jordan compared with different countries.

Fatal poisoning with alcohol and drugs in the Greater Amman County

A study of fatal poisoning due to alcohol and drugs was carried out, to examine the mortality resulting from alcohol and drugs in the Greater Amman County, Jordan. A retrospective review of all autopsy records and certified deaths issued by the Department of Forensic Medicine at Jordan University Hospital in the greater Amman county was undertaken. During the 18 years (1978-1996) 6109 postmortem cases were performed in our department. A total of 60 cases were identified and analyzed according to age, race, sex, manner of death of the victims along with blood alcohol concentration, the drug detected at autopsies, the scene circumstances, and the geographic location of the accident and death.

Investigating the presence of a common drug of abuse (benzhexol) in hair; the Jordanian experience

The analysis of hair is now accepted as a recognised alternative method for the determination of drug misuse. It has several advantages over the biological fluids; blood and urine, including collection of information regarding long-term drug use and determination of compliance with treatment programmes. In Jordan, the abuse of Artane (benzhexol hydrochloride) has been recognised as the most commonly abused drug among Jordanian youths. Hair samples were collected from nine patients (Male 25-55 years, M=39.11, SD=10.53, CV=26.93%). Samples were analysed for the presence of benzhexol and the toxicological analysis revealed the presence of benzhexol in all samples and its concentration ranged from 0.104 to 7.81 ng/mg hair. Solid phase extraction and GC-MS on selective ion storage (SIS) were used for extraction and detection of the drug with papaverine as external standard. The mass detector was operated at selective ion storage (SIS) to monitor the m/z of 98 and 218 for benzhexol and m/z 339 and 324 for the papaverine. The retention times of benzhexol and papaverine were 6.77 and 12.48 min, respectively. The method was linear in the range of 0.5 to 40 ng/mg hair, with a mean coefficient of determination (R2=0.9982). The limit of detection was 0.04 ng/mg. The intra- and inter-day variations were 3.85% and 3.35%. Recovery was found to be above 90%.

Genetic polymorphisms of the CYP3A4, CYP3A5, CYP3A7 and CYP1A2 among the Jordanian population

Cytochromes P450 (CYP450) plays an extremely vital role in oxidation, reduction, and peroxidation of numerous endogenous and exogenous compounds, like drugs and procarcinogens. Mainly, expression occurs in the liver, in varying polymorphic forms. Therefore, proposed as biomarkers of susceptibility to carcinogenicity and toxicity. The objective of this study was to find the allelic frequencies of CYP3A5*2,*3,*4,*5,*6,*7, CYP3A4*1B, CYP3A7*1C and CYP1A2*1C, *1D, *1E, *1F enzymes in Jordanians, and to compare them with other ethnic groups. We used polymerase chain reaction-restriction fragment length (PCR-RFLP) to genotype alleles, and we calculated frequencies using Hardy Weinbergs equation (HWE). Allelic frequencies results were: CYP3A5*2 (0.2%), CYP3A5*3 (86.6%), CYP3A5*6 (1.7%), CYP*3A5*4,*5*7 not detected, CYP3A4*1B (11.7%), CYP3A7*1C (1.7%). Finally 6.5%, 18.2%, 6.0%, 67.3% were the results of CYP1A2*1C, 1D, 1E and 1F, respectively. In conclusion, genotyping method and results of this study can be adopted or used in pharmacotherapy, toxicity and carcinogenic studies in Jordan.

Alcohol and Psychoactive Drugs in Road Traffic Fatalities within Northern District of Amman

Objective: Several studies have demonstrated that alcohol and psychoactive drugs consumption are important risk factors underlying fatal accidents. This paper presents a prospective and a pilot study for toxicological findings in road traffic accident fatality victims within the northern district of Amman in order to have an overall picture of the occurrence of these substances in the victims in Jordan. Method: Over a two-year period (2004-2005), 400 autopsies were conducted at Jordan University Hospital in which 94 (23.5%) were road traffic accident fatality victims. Biological specimens from these victims were collected. Toxicology screens for psychoactive drugs and alcohol were conducted on these specimens and the results were analyzed according to age, sex, and victim’s status. Results: The data showed that ethanol and psychoactive drugs were positive in 72.3%, n = 68 and for ethanol alone 23.4%, n = 22. The majority of victims were the driver (n = 29) and the highest percentage of positive findings occurred among them (20.2%, n=19). Males in the age group 19-29 were mostly involved (35.7%). Detected psychoactive drugs were benzodiazepines, opioids, barbiturates and anti-depressants. None of the collected specimens were positive for illicit cocaine, amphetamines, or cannabis. Conclusions: The results of this study documented the presence of alcohol and psychoactive drugs in road traffic accident victims which could have some association between the uses of these substances and the accident involvement. However, due to some limitations, conclusions require further data collections, cooperation with related parties in Jordan, and utilizing simple and extended toxicological screens.

The Determination of 25-OHVitamin (D2/D3) in Human Serum by Liquid Chromatography Tandem Mass Spectrometry with Comparison to IDS Enzyme Immunoassay

The proper assessment of the status of vitamin D requires the accurate measurement of both 25-OH vitamin D2 and 25-OH vitamin D3, which collectively constitute 25-OH vitamin D, the best indicator of vitamin D status. Currently, numerous assay methods are available for 25-OH vitamin D measurement but their comparability is uncertain. We employed isotope dilution liquid chromatography coupled with tandem mass spectrometry (ID-LC-MS/MS) to quantify 25-OH vitamin D2 and 25-OH vitamin D3 in human serum. Hexadeuterium labeled 25-OH vitamin D3 internal standard was added to calibrators, controls prepared in 6% bovine serum albumin in phosphate buffered saline, and patients’ sera. Zinc sulphate was added to release 25-OH vitamin D metabolites for vitamin D binding protein, followed by a precipitation step withthe addition of acetonitrile. Subsequent online phase extraction by trap column followed by chromatographic separation on a C-8 column using a water/acetonitrile gradient was employed. Detection was performed using Atmospheric Pressure Chemical Ionization (AP-CI) in a Multiple Reaction Monitoring (MRM) mode. The method was linear from 4 to 70 ng/mL. The intra and inter-day CV% were ≤ 10 for both 25-OH vitamin D2 and 25-OH vitamin D3. Recoveries ranged between 39.09 % to 64.31 % for 25-OH vitamin D2 and 30.44 % to 58.66 % for 25-OH vitamin D3, while recoveries from hexadeuterium 25-OH vitamin D3 ranged from 44.11 % to 67.5%.We compared the newly validated LC-MS/MS with a commercial Enzyme Immunoassay from Immunodiagnostic Systems (IDS EIA) in terms of inter-method agreement, correlation, and impact of assay variation on the diagnostic categorization of vitamin D status through the measurement of 182 subjects’ sera. The mean bias % of the IDS EIA relative to the LC-MS/MS was -34.28 ± 10.15 (mean ± std) with 95% CI [-24.13 to 44.43]. The two methods were ingood agreement with reasonable correlation (r²=0.82, P value = 0.000). Inter-method diagnostic categorization was variableand depended on the type of assay method and the applied cut offs used. Cross calibration and standardization of vitamin D assay methods is crucial for proper clinical assessment of vitamin D status. This LC-MS/MS method is reliable and robust for the measurement of both 25-OH vitamin D2 and 25-OH vitamin D3 in human serum.