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Liquid chromatographic-atmospheric pressure chemical ionization mass spectrometric analysis of opiates and metabolites in rat urine after inhalation of opium.

To examine the urinary excretion of opiates and their metabolites following inhalation exposure of rats to opium, analytical procedures for the simultaneous determination of the compounds in opium, the vapor derived by the volatilization of opium and the urine of rats exposed to the opium vapor were developed using liquid chromatography-atmospheric pressure chemical ionization mass spectrometry (LC-APCI-MS). Seven compounds were determined in the opium, namely morphine, codeine, thebaine, noscapine, papaverine, meconic acid and meconin. All seven were extracted with 2.5% acetic acid solution and subjected to LC-APCI-MS analysis. The separation was performed on an ODS column in acetonitrile-50 mM ammonium formate buffer (pH 3.0) using a linear gradient program and quantitative analysis was carried out in the selected ion monitoring mode ([M+H](+)). For the analysis of the volatilization of opium, the opium (1 g) was added to a glass pipe, which was then heated at 300 degrees C for 20 min. Negative pressure (air flow-rate; 300 ml/min) was used to draw the vapor through a series of glass wool and methanol traps. The total amount of each compound in the vapor was estimated by measurement of the compounds trapped in the glass wool and methanol. Wister rats (n=3) were exposed to the vapor derived from the volatilization system and the urinary amounts (0-72 h) of the six opiates and metabolites including morphine-3-grucronide (M3G) and morphine-6-grucronide (M6G) were measured after solid-phase extraction. The calibration curves for those compounds in the rat urine were linear over the concentration range 10-500 ng/ml. The recoveries for each analyte from the rat urine sample spiked with standard solution were generally greater than 80%, and the relative standard deviation for the analytical procedure was less than 8% with the exception of meconin. After inhalation exposure of rats to opium, M3G (5.45-14.38 micro g), morphine (2.27-4.65 micro g), meconin (0.54-1.85 micro g), codeine (0.54-1.85 micro g), noscapine (0.34-0.40 micro g) and papaverine (0.01-0.04 micro g) were detected in the urine over 72 h. However, only trace levels of thebaine were observed despite it being one of the major alkaloids found in the opium. On the other hand, a relatively large amount of meconin was detected in the vapor and the urine as compared with the opium. It is suggested that the presence of meconin in biological fluids could be indicative of opium ingestion by inhalation.

Administration, Inhalation↗

[History of opium poppy and morphine].

Opium has been known for millennia to relieve pain and its use for surgical analgesia has been recorded for several centuries. The Sumerian clay tablet (about 2100 BC) is considered to be the world's oldest recorded list of medical prescriptions. It is believed by some scholars that the opium poppy is referred to on the tablet. Some objects from the ancient Greek Minoan culture may also suggest the knowledge of the poppy. A goddess from about 1500 BC shows her hair adorned probably with poppy-capsules and her closed eyes disclose sedation. Also juglets probably imitating the poppy-capsules were found in that period in both Cyprus and Egypt. The first authentic reference to the milky juice of the poppy we find by Theophrastus at the beginning of the third century BC. In the first century the opium poppy and opium was known by Dioscorides, Pliny and Celsus and later on by Galen. Celsus suggests the use of opium before surgery and Dioscorides recommended patients should take mandrake (contains scopolamine and atropine) mixed with wine, before limb amputation. The Arabic physicians used opium very extensively and about 1000 AD it was recommended by Avicenna especially in diarrhoea and diseases of the eye. Polypharmacy, including a mixture of nonsensical medications were often used. Fortunately for both patients and physicians many of the preparations contained opium. The goal was a panacea for all diseases. A famous and expensive panacea was theriaca containing up to sixty drugs including opium. Simplified preparations of opium such as tinctura opii were used up to about 2000 in Denmark. In the early 1800s sciences developed and Sertürner isolated morphine from opium and was the founder of alkaloid research. A more safe and standardized effect was obtained by the pure opium. Several morphine-like drugs have been synthesized to minimize adverse effects and abuse potential. Opioid receptors were identified and characterized in binding assays and their localization examined. However, the complexity of the system including interaction with several neurons and transmitters indicate the goal of nonaddictive opiates to be elusive. Combination therapy, innovative delivery systems and long-acting formulations may improve clinical utility.

Ancient Lands↗

The GC-MS detection and characterization of reticuline as a marker of opium use.

Reticuline (a precursor of opium alkaloids) was detected and characterised as its trimethylsilyl ethers, acetyl esters and methyl ethers by GC-EIMS and GC-CIMS in opium and the urine of opium users after hydrolysis by acid or beta-glucuronidase as coextractive of morphine. Because this compound cannot be detected in heroin and poppy seeds, it is suggested as a differentiating marker between opium and heroin use, opium and poppy seeds use, or opium and "pharmaceutical" codeine use in cases when opiate use has been confirmed by detection of morphine and codeine in the urine. As well as being a constituent of opium, reticuline in the urine of opium users may also result from the metabolic demethylation of the three other benzyltetrahydroisoquinoline opium alkaloids: codamine, laudanosine and laudanine.

Alkaloids↗

The GC-MS detection and characterization of reticuline as a marker of opium use.

Reticuline (a precursor of opium alkaloids) was detected and characterised as its trimethylsilyl ethers, acetyl esters and methyl ethers by GC-EIMS and GC-CIMS in opium and the urine of opium users after hydrolysis by acid or beta-glucuronidase as coextractive of morphine. Because this compound cannot be detected in heroin and poppy seeds, it is suggested as a differentiating marker between opium and heroin use, opium and poppy seeds use, or opium and "pharmaceutical" codeine use in cases when opiate use has been confirmed by detection of morphine and codeine in the urine. As well as being a constituent of opium, reticuline in the urine of opium users may also result from the metabolic demethylation of the three other benzyltetrahydroisoquinoline opium alkaloids: codamine, laudanosine and laudanine.

Alkaloids↗

Enhanced prolactin levels in opium smokers.

In Iran, opium is smoked for pleasure or as a medication by some people. It is a complex mixture of 40 different alkaloids, including morphine and codeine along with many impurities. Although it is well established that opioids or tobacco affect many physiological functions in humans, to our knowledge there has been no specific study looking at these effects in opium smokers. To assess that, we investigated the circulating levels of prolactin, TSH, LH, FSH and testosterone in male opium smokers who also smoke cigarettes (n=23, aged 28.4+/- 4.1 years), and comparing this with the corresponding values for nicotine abusers (n=12, 15-25 cigarettes/day) or a healthy control group (n=20) of the same age. Our results showed that 86.96% of the opium-dependent and 41.67 % of the nicotine-dependent group displayed high prolactin values (p<0.002). In addition, there was a positive correlation between the dose of opium and the plasma prolactin level of opium dependents (p=0.748, p<0.001). Low FSH was detected in 43.48% of the opium smokers and 50% of the cigarette smokers (p<0.001) with normal LH and testosterone levels. TSH of the opium smokers was also lower than that of the other two groups (p<0.002). In conclusion, the present data indicate that chronic opium and cigarette smoking may synergistically influence pituitary hormone production through the effects on neuropeptides produced either locally or systemic.

Adult↗

Opium and heroin addicts in Laos. II. A study of matched pairs.

Fifty-one Asian heroin addicts in Laos were matched for sex, ethnicity, and age with 51 opium addicts. All subjects were voluntary patients at a treatment facility for addicts. The two groups were compared for demographic characteristics, past narcotic history, current narcotic use, and readmission within 1 year following discharge from treatment. Heroin addicts took more doses of drug per day, spent more money per day on narcotic drugs, required higher detoxification doses of methadone, and sought treatment much sooner than did opium addicts. The two groups did not differ for duration of narcotic use prior to becoming addicted, or for rate of readmission following treatment. Demographic differences in occupation and employment reflected the urban distribution of heroin addicts, and the mixed urban-rural residence of opium addicts. These data suggest that heroin is not per se more or less apt to produce addiction (i.e., is not more "addictogenic") as compared to opium. The type of narcotic drug also does not appear to be an important factor in determining treatment outcome. However, heroin does appear to be more "pathogenic" than opium, since heroin addicts sought treatment much sooner than did opium addicts. This may have been due to economic factors (i. e., heroin addicts took more doses per day, spent more time in phases of intoxication and withdrawal, and spent less time in the middle phase with work and other coping behaviors). Opium addiction is not a "benign" or "social" form of addiction. In comparison to heroin, however, opium does cost less, requires fewer doses per day, and has a less toxic withdrawal (at least in the initial phase). Moreover, opium apparently takes longer to produce life crises that motivate the addict to seek treatment.

Adolescent↗

Effect of opium smoking on concentrations of carcinoembryonic antigen and tissue polypeptide antigen.

Previous studies have related opium and its pyrolysates to the risk of developing certain cancers. The aim of this work was to evaluate the clinical usefulness of determining carcinoembryonic antigen (CEA) and tissue polypeptide antigen (TPA) levels in habitual opium smokers. Serum CEA concentrations were measured in 128 opium smokers and in 44 controls of cigarette only smokers and 47 normal non-smokers by an EIA-based assay. TPA levels were also determined in serum and urine of a subgroup in the study population. The results indicated that serum CEA concentrations are higher in opium smokers than in healthy tobacco smokers (p = 0.004) and non-smokers (p = 0.001). The amount of opium used correlated with the serum CEA level (r = 0.276, p < 0.0001). The mean urine and serum TPA levels of the opium-addicted population were also higher than that of the non-smoking control group, but the differences were not statistically significant. We conclude that opium smoking is associated with elevated serum CEA levels. Therefore, for management of opium users with neoplastic diseases, increased levels of serum CEA should be viewed with caution to avoid misdiagnosis.

Adult↗

Source identification of Indian opium based on chromatographic fingerprinting of amino acids.

Total and free pool of amino acids was determined in Indian opium samples using liquid chromatography (LC) with post-column opthalaldehyde derivatization followed by its fluorimetric detection. The limit of detection (LOD) was found to be in the range of 2-10 pmol with a signal to noise ratio of 3:1 and limit of quantitation (LOQ) was found to be in the range of 7-31 pmol with a signal to noise ratio of 10:1. The recovery of amino acids was found to be in the range of 86-103%. A total of 124 Indian opium samples were collected from the states of Madhya Pradesh (MP), Uttar Pradesh (UP) and Rajasthan (Raj), covering 14 licit opium growing divisions of India were chromatographically fingerprinted for the presence of various amino acids. The amino acids identified in sample hydrosylate included D, T, S, S, G, A, V, I, L, Y, F, H, K and R, while the analysis of free pool of amino acids (80% aqueous ethanol extract) indicated the presence of D, T, S, E, A, V, I, L, Y, H, K respectively. Multiple discriminant analysis was applied to the quantitative total amino acid data to determine an optimal classifier in order to evaluate the source of Indian opium. The foremost amino acid variables that accounted for the true discrimination were identified as D, E, G, A, F and K in evaluating the geographical origin of Indian opium and the predictive value based on the discriminant analysis was found to be 90% in relation to the source of opium samples. Chemometrics performed with amino acid analytical data was used successfully in discriminating the licit opium growing divisions of India into three major groups, viz. groups I, II and III. The methodology developed may find wide application in forensic analysis.

Amino Acids↗

Opium and risk of laryngeal cancer.

OBJECTIVES/HYPOTHESIS: The study was performed to investigate the possible association between opium dependency and laryngeal cancer. STUDY DESIGN: A hospital-based, group-matched, case-control study was presented. METHODS: Ninety-eight patients with laryngeal cancer and 312 age- and gender-matched control subjects were selected at the otorhinolaryngology ward of a referral university-affiliated hospital. Data on cigarette smoking and alcohol and opium dependency were collected before surgery through semi-structured interview. RESULTS: The crude odds ratios of laryngeal cancer were 15.07 (95% confidence interval [CI], 6.92-32.8 [P <.0001]) for cigarette smoking, 21.55 (95% CI, 10.54-44 [P <.0001]) for opium dependency, and 1.84 (95% CI, 1.008-3.38 [P <.048]) for male gender. Because of strong associations, a logistic regression model was prepared; the odds ratio for gender in the final model was 0.87 (95% CI, 0.39-1.92 [P =.11]). According to the results, it seemed that gender was not an independent risk factor for laryngeal cancer. Also, the adjusted odds ratios for smoking (5.21) (95% CI, 2.33-11.67 [P <.002]) and opium dependency (10.74) (95% CI, 5.76-20.02 [P <.002]) were lower than the crude odds ratios, but both ratios were significant. The mean patient ages were 55.1 years (SD = 12.05 y) in opium-dependent patients and 65.6 years (SD = 12.8 y) in opium-nondependent patients (P =.01). CONCLUSION: The results of the study suggest that opium dependency is not only an independent possible risk factor for laryngeal cancer but also significantly increases the likelihood of developing of the disease at a younger age.

Aged↗