UHPLC-MS/MS quantitation of porphyroxine in opium and application of porphyroxine-acetylated products as signature markers for heroin.

Porphyroxine, a trace alkaloid in opium, was identified in the early 1800s and isolated/characterized in the 1960s. Recently, two significant porphyroxine-related byproducts found in the acidic and neutral extracts of illicit heroin were characterized by this laboratory as the N-acetyl-O14 -desmethyl-epi-porphyroxine (B) and N,O8 -diacetyl-O14 -desmethyl-epi-porphyroxine (C). The prevalence of the B and C compounds has been consistent in the following order of abundance for the thousands of authentic heroin samples analyzed: Southwest Asia (SWA) > South America (SA) > Southeast Asia (SEA) > Mexico (MEX). In this research, a rapid and efficient ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method was developed to determine the content of porphyroxine and five primary alkaloids (morphine, codeine, thebaine, noscapine, and papaverine) in opium after extraction with methanol/water (50/50). The method was validated in terms of linearity, accuracy, recovery, and precision for porphyroxine. The limit of quantitation (LOQ) for porphyroxine was 2.5 ng/mL. The developed method was successfully applied to a total of 114 authentic opium samples from the major poppy-growing regions. The amount of porphyroxine was determined at the level of part per thousand (‰) and the relative concentrations to morphine were in the range of 1x10-4 and 1x10-2 with an order of SWA > SEA, SA > MEX for its average abundance, which is consistent with the order of the average abundance of its acetylated products (B, C) in illicit heroin. This study reveals the significance of porphyroxine and its acylated compounds in classifying heroin and opium samples to major geographical regions of production.

Kaurene as the major constituent of the essential oils of the narcotic plant, Khat (Catha edulis Forsk).

Khat (Catha edulis Forsk) is a narcotic plant which contains significant amounts of amphetamines, like alkaloids. Herein, analysis of the essential oil composition showed that Khat has useful volatile chemicals in addition to its alkaloids. Results indicated that among 35 identified constituents including mono and sesquiterpenes, the diterpene kaurene, comprises the major part of the essential oil, around 50 percent of total. Kaurene is known as a potent biological agent for the treatment of cancer patients. The presence of kaurene at high levels indicates that the essential oil of Catha edulis can potentially be more effectively exploited rather than its narcotic stimulant amphetamine-like alkaloids.

DARK Classics in Chemical Neuroscience: Opium, a Historical Perspective.

Opium is the latex from the opium poppy Papaver somniferum L., which humankind has utilized since ancient Mesopotamia all the way to modern times. Opium used to be surrounded in divine mystery or magic-like abilities and was given to cure a wide variety of diseases until its analgesic, antitussive, and antidiarrheal properties were understood, the resulting alkaloids were isolated, and their structure and properties unmasked. Opium went from being sold in any store front in the form of pills or tinctures with no prescription necessary for purchase or smoked in an opium den down the street, to then bringing about consumer advocacy and the right to know what is in a medication. Legislation was created to limit the prescribing and selling of medications to doctors and pharmacists as well as outlawing opium dens and smoking opium. This review focuses primarily on the uses of opium throughout history, the isolation of the principle alkaloids, and their structure elucidation.

Development and validation of an LC-MS/MS method for the simultaneous analysis of 28 specific narcotic adulterants used in dietary supplements.

The purpose of this study was to develop a method to analyse the concentration of multiple illegal narcotics present in dietary supplements. To this end, we established and optimised a procedure using LC-MS/MS simultaneously to analyse 28 narcotic compounds in various forms of dietary supplements, including powders, tablets, liquids and capsules. In addition, candy and cookies that have also had detected cases of adulteration were also analysed. The specificity, linearity, accuracy, precision, limit of detection (LOD), limit of quantitation (LOQ), stability and recovery for these methods were validated accordingly. The LOD and LOQ of the LC-MS/MS ranged from 0.01-50.0 to 0.03-100 ng g(-1), respectively. The linearity of these results was good (r(2) > 0.99), with intra- and inter-day precision values of 0.2-5.2% and 0.2-4.8%, respectively. Further, the intra- and inter-day accuracies of this method were 97.0-103.4% and 94.6-103.1%, respectively. The stability RSD was less than 7.8%. The mean recovery for this LC-MS/MS procedure was 81.1-117.4%, with an RSD less than 9.8%. Following the validation of our method, we analysed 47 commercially available dietary supplements obtained in Korea. Whilst none of these samples had detectable amounts of the 28 specified narcotic adulterants, our novel LC-MS/MS procedure can be utilised comprehensively and continually to monitor illegal drug adulteration in various forms of dietary supplements.

[The clinical and X-ray classification of osteonecrosis of the low jaw].

OBJECTIVE: To elaborate a clinical and X-ray classification of osteonecrosis of the low jaw in people with desomorphine or pervitin addiction. MATERIAL AND METHODS: Ninety-two patients with drug addiction who had undergone orthopantomography, direct frontal X-ray of the skull, and multislice computed tomography, followed by multiplanar and three-dimensional imaging reconstruction were examined. One hundred thirty four X-ray films and 74 computed tomographic images were analyzed. RESULTS: The authors proposed a clinical and X-ray classification of osteonecrosis of the low jaw in people with desomorphine or pervitin addiction and elaborated recommendations for surgical interventions on the basis of the developed classification. CONCLUSION: The developed clinical and X-ray classification and recommendations for surgical interventions may be used to treat osteonecroses of various etiology.

Narcotic alkaloids of four papaver species from Iran.

Four native Papaver species of Iran, i. e. P. glaucum, P. tenuifolium, P. dubium and P. fugax, were collected from their natural habitat and subjected to HPLC analysis for determination of their morphine, codeine and thebaine content. P. dubium and R. glaucum contained all of the three mentioned narcotic alkaloids, while morphine was not found in P. fugax, and P. tenuifolium was free from codeine.

Involvement of mu-opioid receptors in antinociception and inhibition of gastrointestinal transit induced by 7-hydroxymitragynine, isolated from Thai herbal medicine Mitragyna speciosa.

7-hydroxymitragynine, a constituent of the Thai herbal medicine Mitragyna speciosa, has been found to have a potent opioid antinociceptive effect. In the present study, we investigated the mechanism of antinociception and the inhibitory effect on gastrointestinal transit of 7-hydroxymitragynine, and compared its effects with those of morphine. When administered subcutaneously to mice, 7-hydroxymitragynine produced antinociceptive effects about 5.7 and 4.4 times more potent than those of morphine in the tail-flick (ED50=0.80 mg/kg) and hot-plate (ED50=0.93 mg/kg) tests, respectively. The antinociceptive effect of 7-hydroxymitragynine was significantly blocked by the mu1/mu2-opioid receptor antagonist beta-funaltrexamine hydrochloride (beta-FNA) and the mu1-opioid receptor-selective antagonist naloxonazine in both tests. Thus, 7-hydroxymitragynine acts predominantly on mu-opioid receptors, especially on mu1-opioid receptors. Isolated tissue studies further supported its specificity for the mu-opioid receptors. Further, 7-hydroxymintragynine dose-dependently (ED50=1.19 mg/kg, s.c.) and significantly inhibited gastrointestinal transit in mice, as morphine does. The inhibitory effect was significantly antagonized by beta-FNA pretreatment, but slightly antagonized by naloxonazine. The ED50 value of 7-hydroxymitragynine on gastrointestinal transit was larger than its antinociceptive ED50 value. On the other hand, morphine significantly inhibits gastrointestinal transit at a much smaller dose than its antinociceptive dose. These results suggest that mu-opioid receptor mechanisms mediate the antinociceptive effect and inhibition of gastrointestinal transit. This compound induced more potent antinociceptive effects and was less constipating than morphine.

Comparative qualitative and quantitative determination of alkaloids in narcotic and condiment Papaver somniferum cultivars.

In the present study morphinan, tetrahydrobenzylisoquinoline, benzo[c]phenanthridine, and phthalideisoquinoline alkaloids were determined qualitatively and quantitatively by HPLC and LC-MS analysis in tissues of the Tasmanian Papaver somniferum L. elite cultivar C048-6-14-64. The data were compared with the results from the low-morphine cultivar "Marianne". In the elite cultivar, 91.2% of the latex alkaloids consist of the three pharmaceutically most valuable alkaloids: morphine, codeine, and thebaine. In the root system, the major alkaloids are sanguinarine/10-hydroxysanguinarine and dihydrosanguinarine/10-hydroxydihydrosanguinarine. In the stems and leaves of C048-6-14-64, the same alkaloids were measured as in the latex. In the stems, a gradient in relative total alkaloid content from the top downward toward the roots was observed. The concentration of morphine was decreasing toward the roots, whereas an increasing gradient from the upper to the lower stem parts was detected for codeine. The relative total alkaloid concentration in leaves remained constant; no gradient was observed. The cultivar "Marianne" displayed a shifted pattern of alkaloid accumulation and reduced levels of total alkaloid. In the condiment cultivar, 80.5% of the alkaloids of the latex consisted of the two phthalideisoquinoline alkaloids narcotoline and noscapine. Only 18.8% of the relative total alkaloid content were morphinan alkaloids. In contrast to the narcotic cultivar, in which the benzo[c]phenanthridines in roots dominated over the morphinan and tetrahydrobenzylisoquinoline alkaloids, the concentration of benzo[c]phenanthridines in "Marianne" was similar to that of morphinan and tetrahydrobenzylisoquinoline alkaloids. These data suggest a differential alkaloid regulation in each cultivar of P. somniferum.

Investigation of heroin profiling using trace organic impurities.

The acidic and neutral impurities in heroin samples (46 Purified South-East Asian (PSEA) and 8 non-PSEA samples) were analysed using gas chromatography. Mass spectral detection allowed the construction of a comprehensive library of over 649 impurities. A variety of statistical procedures were used to select 70 viable profiling parameters. Cluster analyses were used to investigate the similarities between samples using organic parameters. A blended PSEA profile was constructed for comparison with samples of unknown origins. Resolution between samples of PSEA/non-PSEA origins was demonstrated with a selection of 25 (18 continuous and 7 dichotomised) discriminating factors. Several predictive logistic models were created using up to 18 parameters, explaining 85-100% of the variation in the experimental data.

Genetic and alkaloid analysis of Papaver species and their F1 hybrid by RAPD, HPLC and ELISA.

Total DNA was extracted from the leaves of a F1 hybrid and its parents, Papaver bracteatum Lindle and P. pseudo-orientale Medw. Analysis of random-amplified polymorphic DNA (RAPD) using ten arbitrary oligonucleotide 10-mers, showed that F1 hybrid was confirmed to be genetically intermediate of both parental plants compared with the genetic distance between F1 hybrid and individual parents. Furthermore, the comparison of the band patterns between a F1 hybrid, P. bracteatum and P. pseudo-orientale clearly showed that part of the bands of both parents were induced into a F1 hybrid. The content of thebaine was determined by HPLC and ELISA used anti-thebaine monoclonal antibody.

Pharmaceutical applications of micelles in chromatography and electrophoresis.

This review surveys the use of micelles as separation media in chromatography and electrophoresis. Applications to pharmaceuticals whose molecular masses are relatively small are focused on in this review. In high-performance liquid chromatography (HPLC), chromatography using micelles and reversed-phase stationary phases such as octadecylsilylized silica gel (ODS) columns is known as micellar liquid chromatography (MLC). The main application of MLC to pharmaceutical analysis is the same as in ion-pair chromatography using alkylsulfonate or tetraalkylammonium. In most cases, selectivity is much improved compared with other short alkyl chain ion-pairing agents such as pentanesulfonate or octanesulfonate. Direct plasma/serum injection can be successful in MLC. Separation of small ions is also successful by using gel filtration columns and micellar solutions. In electrophoresis, especially capillary electrophoresis (CE), micelles are used as pseudo-stationary phases in capillary zone electrophoresis (CZE). This mode is called micellar electrokinetic chromatography (MEKC). Most of the drug analysis can be performed by using the MEKC mode because of its wide applicability. Enantiomer separation, separation of amino acids and closely related peptides, separation of very complex mixtures, determination of drugs in biological samples etc. as well as separation of electrically neutral drugs can be successfully achieved by MEKC. Microemulsion electrokinetic chromatography (MEEKC), in which surfactants are also used in forming the microemulsion, is successful for the separation of electrically neutral drugs as in MEKC. This review mainly describes the typical applications of MLC and MEKC for the analysis of pharmaceuticals.

Opium revisited: a brief review of its nature, composition, non-medical use and relative risks.

Unlike the pure opioids such as morphine and heroin, opium is a complex and variable mixture of substances reflecting differences in both the starting material and the traditional practices of the regions in which it is produced. Analytical methods have improved greatly in recent years, to the point that the source of a preparation can often be identified by its opioid content and its impurities. Daily amounts used, both by smoking and by mouth, vary widely from less than a gram to 30 g, equivalent to 75-3000 mg of morphine. The effects of opium are essentially those of morphine but unexpected toxicities, such as oesophageal cancer associated with "dross opium" and polyneuropathy due to deliberate addition of arsenic, are problems in some specific regions. Prevalence of use in different areas and countries is governed by the same factors of ease of availability, price and social acceptance that apply to the use of alcohol and other drugs in western countries. The risk of addiction to opium smoking appears to be somewhat less than to parenteral use of heroin, but appreciably greater than to alcohol. Even in countries where its use is traditional, opium smoking carries substantial risks of harm to health and social functioning.