ABSTRACT
The paper reports the establishment of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) for simultaneous analysis of 11 opiates in hair samples, and the study of presence of opiates in the hair of active heroin addicts. About 20 mg of decontaminated and pulverized hair sample was hydrolyzed with buffer solution for 30 min, in the presence of morphine-d3 and acetylmorphine-d6 used as internal standards, and then extracted with the mixture of dichlormethane and isopropanol, separated by the Allure PFP propyl column with a mobile phase consisting of acetonitrile and 20 mmol L(-1) ammonium acetate buffer, and then analyzed by LC-MS/MS. Multiple reaction monitoring (MRM) mode was used to analyze 11 opiates. Eleven opiates showed a fairly good linearity over the corresponding range (r > 0.996 0). The detection limits were less than 0.05 ng mg(-1). The recoveries were between 47.2% and 110%, and the deviations of intra- and inter-day precision were less than 14%. Heroin, acetylmorphine, morphine, codeine, acetylcodeine and hydrocodone were detected in hair samples of 21 herion addicts. The developed method shows high sensitivity and selectivity, and is suitable for the simultaneous analysis of 11 opiates in hair samples and identify legal and illegal use of opiates.
Subject(s)
Humans , Analgesics, Opioid , Chromatography, Liquid , Methods , Codeine , Hair , Chemistry , Heroin , Hydrocodone , Limit of Detection , Morphine , Morphine Derivatives , Sensitivity and Specificity , Substance Abuse Detection , Methods , Tandem Mass Spectrometry , MethodsABSTRACT
OBJECTIVE@#Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was developed for the determination of opiates in biological samples according to the emerging problem in drugs abuse.@*METHODS@#Opiates such as heroin, 6-acetylmorphine, morphine, codeine, acetylcodeine, hydrocodone and hydromorphone were isolated from human blood, urine, oral fluid and hair using a simple extraction and consequently analyzed using LC-MS/MS. The method was evaluated by real cases.@*RESULTS@#The mobile phase give the optimum separation for opiates. The detection limit of morphine in urine with dilution and liquid-liquid extraction and in hair is 10ng/mL, 0.01 ng/mL and 0.01 ng/mg, respectively.@*CONCLUSION@#The method is simple and rapid, offering superior sensitivity and selectivity for opiates. The target compounds comprising hydrocodone and hydromorphone enlarge the applied area.
Subject(s)
Humans , Chromatography, Liquid , Codeine/analysis , Forensic Medicine/methods , Hair/chemistry , Hydrocodone/analysis , Hydromorphone/analysis , Morphine/analysis , Narcotics/analysis , Reproducibility of Results , Saliva/chemistry , Substance Abuse Detection/methods , Tandem Mass SpectrometryABSTRACT
The proliferation of smooth muscle cells in the arterial wall [VSMC] is considered to play a key role in the development of atherosclerosis. To investigate the possible contribution of "stress" [experimentally induced] to this process, blood from healthy volunteers, ages 21 to 65, screened to exclude major risk factors for coronary heart disease, was assayed for mitogenic activity after the subjects were exposed to one of 2 "stress" conditions. These consisted of a cognitive task with superimposed verbal harassment [group 1], and the cognitive task without harassment [group 2]. Mitogenic activity was determined by studying the growth stimulatory effects of PDGF-depleted plasma derived serum [PDS] from "stressed" subjects added to cultured VSMC, as measured by incorporation of radioactive thymidine into DNA or increase in cell number. In addition, changes in the steady state of the mRNA for the c-myc protooncogene were also assayed in VSMC by Northern blot analysis, using sera showing the greatest differential "pre/post stress" mitogenic activity. Blood pressure [BP], heart rate [HR], cortisol, and serum total and HDL cholesterol were also evaluated. All measurements were made immediately before [baseline] and after a 30 min interval. Analysis of the data revealed that there were 33% of subjects in group 1 with an increase of thymidine incorporation 15% or greater than baseline, versus 21% in group 2. The average increases were 45% and 30%. A higher percentage [35-42%] of subjects in group 1 responded with increases in systolic and diastolic blood pressure, compared to subjects in group 2 [15-20%]; the average in blood pressure was 10-15%. Similarly, more subjects [52%] in group 1 had an elevated [average 10-15%] serum cortisol, compared to the 42% in group 2 subjects. HR, total HDL cholesterol showed slight changes only. These results suggest that psychoactive factors may affect cardiovascular systems via rapid elicited rises in serum mitogenic activity for VSMC