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Khat belongs to the family Celastraceae, genus Catha, and species edulis. More than 200 compounds have previously been identified in Khat leaves, including: 40 alkaloids, terpenoids and sterol, flavonoids, glycosides, tannins, amino acids, vitamins and minerals. Researchers have spent their effort and time merely on study of the alkaloidal components (mainly the stimulant agents, cathinone and cathine) of Khat both qualitatively and quantitatively. The two principal Khat stimulant compounds, cathinone and cathine, by now are well established. But, on the contrary, previous studies on the non-alkaloidal constituents of the plant were limited. The objective of this work was therefore to isolate and characterize compounds from non-alkaloidal fractions of the plant’s leaves. In this work, two non-alkaloidal compounds (KNA-1 and KNA-2) were isolated and characterized from the acid-EtOAc extract of fresh and oven-dried leaves of Khat (Catha edulis). From the present study, it is possible to conclude that investing more effort and time on searching additional non-alkaloidal principles from the leaves of Khat is so necessary. And further works could be done in the future to isolate extra non-alkaloidal compounds from the leaves and other parts of Khat and evaluate their biological activity.

Background Although psychostimulants apparently do cross the BBB, it is poorly understood how these hydrophilic and positively charged molecules can pass the blood-brain barrier (BBB). That may be mediated by a genetically still uncharacterized H+/OC antiporter with high activity at the BBB. Methods We studied the uptake of 16 psychostimulants and hallucinogens with hCMEC/D3 cells using the prototypic inhibitor imipramine (cis-inhibition), exchange transport with diphenhydramine and clonidine (trans-stimulation), proton dependency of the uptake, and we characterized the concentration-dependent uptake. Results Cell uptake of methylenedioxyamphetamines, amphetamines and dimethyltryptamine (DMT) were strongly inhibited (to about 10% of the controls) by imipramine and diphenhydramine, whereas uptake of cathine was only weakly inhibited and mescaline not significantly. Amphetamine, methylamphetamine, para-Methoxy-N-methylamphetamine (PMMA), Methylenedioxymethamphetamine (MDMA), phentermine and DMT exhibited the highest exchange after preloading with diphenhydramine with only 5.5%, 5.2%, 7.8%, 6%, 1.9%, 7.6% remaining in the cells. Less and no exchange were seen with cathine and mescaline, respectively. Dependence on intracellular pH was most pronounced with the methylendioxyamphetamines while uptake of cathine, DOI and cocaine were only moderately affected and mescaline not at all. Conclusion Except for mescaline, all psychostimulants studied here were substrates of the H+/OC antiporter, implicating a strong need for a better characterization of this transport protein.

Background Molecular docking is commonly used in the study of small-molecule–protein interaction. Amphetamine (AMPH), cathine (CATHI), and cathinone (CATHO) bind in silico with monoamine oxidase B (MAO B) with relatively low affinity. Objective The purpose of the study was to determine the binding modes of AMPH, CATHI, and CATHO with the enzyme MAO B. Molecular docking software, AutoDock 4.2, was used to study the interaction of these ligands with MAO B. Materials and methods Molecular docking was performed using the molecular docking software AutoDock, version 4.2. graphic user interface, AutoDock Tools, was used to prepare protein and ligands for docking. Results The estimated ΔG of binding (free energy of binding) was −6.25 kcal/mol for AMPH, −6.05 kcal/mol for CATHI, and −6.24 kcal/mol for CATHO. Respective Ki (inhibitor constant) values were 2.61×10−5 M, 3.69×10−5 M, and 2.69×10−5 M. All three ligands show similar interactions within active site which include hydrophobic and hydrogen-bonding interactions. When compared with binding affinity of known inhibitors of MAO, safinamide, the binding affinity of these inhibitors is substantially less. But, they bind in the same active site and share some common interactions with active site residues. Similarity in binding modes of AMPH, CATHI, and CATHO is due to their similar structures. Conclusion It can be speculated that nervous stimulant activity of AMPH, CATHI, and CATHO could be due to inhibition of MAO. This enzyme catalyzes the oxidative deamination of monoamine neurotransmitters such as serotonin and dopamine and reduces the level of these neurotransmitters. Inhibition of this enzyme might result in more sustained levels of these neurotransmitters. Our docking study shows that AMPH, CATHI, and CATHO inhibit MAO B with free energy ranging from −6.05 to −6.25 kcal/mol and Ki ranging from 2.61×10−5 to 3.69×10−5 M.

Objectives Khat (Catha edulis) is a stimulant plant, and it is abusive to induce euphoria, alertness and activity. Concomitant use of medications and khat chewing predisposes to the appearance of drug interactions result in treatment failure or toxicity. This study determined the changes in the urinary inorganic profile in adult healthy males who are chewing khat compared with non-khat chewer males. Methods A total of 40 adult non-smoker healthy males (20 khat chewer and 20 non-khat chewer) aged 24–30 years were selected. Khat chewer samples were positive for cathinone and cathine and negative for other drug of abuse, while non-khat chewer samples were negative for drug of abuse include cathinone and cathine. Samples were selected according to their results in immunoassay and gas chromatography mass spectrometry (GCMS) analysis. Cathine and cathinone were confirmed using liquid chromatography mass spectrometry (LC-MSMS) analysis. Inorganic profile includes titanium (Ti), cobalt (Co), copper (Cu), zinc (Zn), cadmium (Cd), and lead (Pb) were determined by using inductively coupled plasma-mass spectrometry (ICP-MS). Results The levels of Ti, Co, Zn, Cd, and Pb in urine were significantly higher among the khat chewer group compared with non-khat chewer. Ti, Cd, Co, Pb and Zn urine levels were 0.5-, 1.5-, 1.15-, 5-, and 8.2-fold higher in the khat chewer group compared to non-khat chewer, respectively. Conclusions We suggested that continuous khat chewing has a long term effect on metabolic pathway of therapeutic drugs that result in toxicity or failure of therapy.

Khat (Catha edulis (Vahl) Endl.) is an evergreen flowering shrub used as a stimulant in many regions worldwide including East Africa, the Arabian Peninsula, Europe, and the United States. Chewing leaves of khat induces excitement and euphoria, which are primarily attributed to two major constituents, cathinone and cathine. Khat also contains other important constituents such as cathedulins. A considerable number of studies reported side effects induced by the khat extracts to both embryos and adults. These include teratogenicity and developmental retardation, oral cancer and ulcers, high blood pressure, and myocardial infarction. So far, little attention has been paid to the effects of khat extracts on the molecular signaling interactions. We aimed in this study to investigate this through evaluating the effects of khat extracts on SKOV3, a human ovarian adenocarcinoma cell line. We show, by in vitro assays, that khat induces several cellular defects including reduced cell size, cell membrane damage, and apoptosis. At high khat extract concentrations, the cell metabolic activity, cell cycle, and cellular proliferation were affected. RT-qPCR analysis showed an increase in the gene expression of the apoptotic marker BAX, the tumor suppressor p53, and the inflammatory cytokine IL-6. Protein expression analysis by immunostaining showed downregulation of β-catenin, E-cadherin, and Ki-67 and upregulation of FZD8 and SPRY2, suggesting that Wnt and FGF signaling were implicated. SwissTargetPrediction in silico analysis showed that khat constituents cathine, cathinone, catheduline K2, and catheduline E5 bind to family A G-protein-coupled receptor, cause many neurological diseases and disorders such as Alzheimer's, schizophrenia, depression, and anxiety, and induce many ovarian cancer-related diseases. The analysis also showed that important signaling pathways such as CREB, Wnt, FGF, IL-6, and ERK/MAPK, and that of the endometrial cancer, and cell cycle were implicated. Upstream regulators of cathine and cathinone were found to potentially target several molecules including interleukin-8, MMP2, PLAU, and micro-RNAs. In conclusion, khat induces significant cellular and molecular changes that could potentially cause a wide range of serious diseases and syndromes. Such an impact could have a heavy burden on the health care system in the countries where khat is consumed.

Psychostimulants are used therapeutically and for illegal recreational purposes. Many of these are inhibitors of the presynaptic noradrenaline, dopamine, and serotonin transporters (NET, DAT, and SERT). According to their physicochemical properties, some might also be substrates of polyspecific organic cation transporters (OCTs) that mediate uptake in liver and kidneys for metabolism and excretion. OCT1 is genetically highly polymorphic, with strong effects on transporter activity and expression. To study potential interindividual differences in their pharmacokinetics, 18 psychostimulants and hallucinogens were assessedin vitrofor transport by different OCTs as well as by the high-affinity monoamine transporters NET, DAT, and SERT. The hallucinogenic natural compound mescaline was found to be strongly transported by wild-type OCT1 with aKmof 24.3 µM and avmaxof 642 pmol × mg protein−1× min−1. Transport was modestly reduced in variants *2 and *7, more strongly reduced in *3 and *4, and lowest in *5 and *6, while *8 showed a moderately increased transport capacity. The other phenylethylamine derivatives methamphetamine,para-methoxymethamphetamine, (-)-ephedrine, and cathine ((+)-norpseudoephedrine), as well as dimethyltryptamine, were substrates of OCT2 withKmvalues in the range of 7.9–46.0 µM andvmaxvalues between 70.7 and 570 pmol × mg protein−1× min−1. Affinities were similar or modestly reduced and the transport capacities were reduced down to half in the naturally occurring variant A270S. Cathine was found to be a substrate for NET and DAT, with the Kmbeing 21-fold and thevmax10-fold higher for DAT but still significantly lower compared to OCT2. This study has shown that several psychostimulants and hallucinogens are substrates for OCTs. Given the extensive cellular uptake of mescaline by the genetically highly polymorphic OCT1, strong interindividual variation in the pharmacokinetics of mescaline might be possible, which could be a reason for highly variable adverse reactions. The involvement of the polymorphic OCT2 in the renal excretion of several psychostimulants could be one reason for individual differences in toxicity.

Background and objective Khat is a plant that contains the alkaloids cathine and cathinone which have some amphetamine-like properties. It is cultivated and it's leaves chewed for their euphoric effect. This study intended to elucidate the effect of khat chewing on blood coagulation by using the International Normalized Ratio (INR) value as a calculable benchmark. Materials and methods In this cohort study, 146 patients with Mechanical Heart Valves (MHV) were assessed for two consecutive visits at one-month intervals. For each visit, the date of surgery, the patient's compliance, the dose of warfarin and the INR reading were assessed by the researcher. Results Out of 146 patients with MHV, the mean age was 33.72±12.43 years (range, 14-65 years); 82 (56.2%) were female and 64 (43.8%) were male. The results revealed that the mean of absolute INR readings was lower in khat-chewers than non-chewers by average 0.2 on the first and second visits (p = 0.038 and 0.002, respectively). Conclusion Khat chewing has a significant coagulant effect. There was a significant decrease in the value of INR for khat chewers patients with MHV when compared to non-khat chewers.

Khat or qat (Catha edulis) is a flowering plant with an Arabic name قات, regularly named as qāt, also is known by various descriptive names, such as Abyssinian tea, Somali tea, Arabian tea, Miraa, Jima, and Kafta in its endemic regions of the Horn of Africa and the Arabian Peninsula. Fresh leaves and tops of khat are chewed or dried and consumed as a tea to attain a state of euphoria and excitement; it also has appetite-reducing effects. Traditionally, khat is used as a socializing habit in Yemen and is also widely cultivated because of its high income. However, in recent years the plant has been reported in England, Wales, Rome, Amsterdam, Canada, Israel, Australia, New Zealand, and the United States. Although it is believed that khat is a relatively low-risk drug, it's associated with an increased risk for various medical complications, including dental and oropharyngeal disease, gastrointestinal, cardiovascular, neuropsychiatric, obstetric, and even can be the cause of cancer. Our goal in this review article is to revise and determine the relationship between chewing khat and its health issues. Additionally, we tried to determine the mechanism involved in health hazards due to consuming the khat.

The consumption of Khat leaves represents an ancient kind of drug abuse mainly observed in Eastern Africa and the Arab Peninsula among adult men. For this purpose, the leaves are directly collected from the shrub "Catha edulis" prior to extensive chewing process. Seizures in Europe are rare, since the leaves have to undergo quick transportation: After a short period of time, the harvested leaves decompose and suffer in decrease of concentration of the active ingredient cathinone, which makes long term transportation difficult. As an alternative, plant material can be dried to increase life period. In the past years, an increasing number of seizures were made by Austrian police, however, the content of cathinone and cathine in dry material is widely unknown. In this work, a seizure of fresh Khat leaves was compared with two seizures of dried material in terms of concentration of cathinone and cathine using LC-MS/MS analysis. For fresh leaves, a purity grade was found to be 0.115-0.158% for cathinone and 0.172-0.192% for cathine, respectively. In contrast, subsequent storage of dried Khat leaves over months led to a dramatic loss of cathinone: Analysis of two seizures revealed that concentration of cathinone dropped to 0.021-0.023%. These findings are intended to serve as a guideline for Justice authorities to estimate the content of the controlled ingredients of Khat leaves in future.

Background: The tradition of khat chewing has been deep-rooted in the African and Arabian Peninsula for centuries. Due to its amphetamine-like psycho-stimulant or euphoric effect, khat has been used by millions in Somalia, Ethiopia, Saudi Arabia and Yemen. The long-term use of khat can induce many major health outcomes, which may be serious and irreversible. Objective: Prolonged use of khat constituents has been associated with different types of cancers such as prostatic, breast and ovarian cancer. However, it has been very difficult to identify the molecular targets involved in khat carcinogenesis that interact with the Khat constituents by in vitro/in vivo experimental tools. Methods: In silico tools were used to predict potential targets involved in the carcinogenesis of khat. Pass on-line prediction server was used for the prediction of a potential molecular target for khat constituents. Molecular Dynamics simulation and MM-GBSA calculation of the predicted target were carried out. Results: Molecular Dynamics simulation and MM-GBSA calculation revealed that among khat constituents, β-sitosterol showed a high binding affinity towards 17β-HSD5. On the other hand, this study highlights for the first time some new interactions, which were observed in the case of cathine, cathinone and nerol during the simulation. Conclusion: In silico molecular dynamic simulation tools were used for the first time to investigate the molecular mechanism of widely used leaves of psychoactive khat (Catha edulis) constituent. The present study provides deep insight to understand the effect of khat constituents involved in the impairment of the reproductive system and its binding to 17β-HSD5. ADMET profiling also suggested that few khat constituents do not fulfill the requirements of the Lipinski rule of five i.e. poor absorption and blood-brain barrier impermeability.

Interpretation of amphetamine-type stimulant (ATS) findings in urine samples can be challenging without chiral information. We present a sensitive enantioselective high-performance liquid chromatography-tandem mass spectrometry method for the quantification of (R)-amphetamine, (S)-amphetamine, (R)-methamphetamine, (S)-methamphetamine, (1R,2R)-pseudoephedrine, (1S,2S)-pseudoephedrine, (1R,2S)-ephedrine, (1S,2R)-ephedrine, (1R,2S)-norephedrine, (1S,2R)-norephedrine, (R)-cathinone, (S)-cathinone, and (1S,2S)-norpseudoephedrine (cathine) in urine. The method was successfully applied to more than 100 authentic urine samples from forensic casework. In addition, samples from a controlled self-administration of (1S,2S)-pseudoephedrine (Rinoral, 1200 mg within 6 days) were analyzed. The results strengthen the hypothesis that (1R,2S)-norephedrine is a minor metabolite of amphetamine and methamphetamine. We suggest cathine and (1S,2R)-norephedrine as minor metabolites of amphetamine racemate in humans. Small methamphetamine concentrations detected in samples with high concentrations of amphetamine could result from a metabolic formation by methylation of amphetamine although in samples with an (R)/(S) ratio for methamphetamine < 1 an additional (previous) (S)-methamphetamine consumption seems likely. Our data suggest that even amphetamine concentrations exceeding methamphetamine concentrations in urine can be caused by the biotransformation of methamphetamine to amphetamine as long as no (R)-amphetamine is detected. However, without chiral information, such findings might be (falsely) assumed as a co-consumption of both substances. Cathinone enantiomers detected in urine samples with high amphetamine concentrations can be interpreted as metabolites of amphetamine. In addition, the results of the self-administration study revealed that both cathinone enantiomers are minor metabolites of (1S,2S)-pseudoephedrine, which is the active ingredient of various medicines used for cold. The enantioselective analysis is a powerful tool to avoid the misinterpretation of ATS findings in urine samples.

Chewing khat leaves has been associated with several adverse health effects, and there are very few case reports of cardiotoxicity, stroke and death resulting from this. In addition, postmortem distribution of cathine and cathinone, active components of khat, are not yet fully clear. This postmortem case report aimed to identify and determine the concentration of cathine and cathinone in different body organs and green chewed plants found in the mouth of the deceased. Immunoassay and non-targeted GC-MS analysis showed that samples were only positive for amphetamine type stimulants. LC-MS/MS quantitative analysis confirmed that samples were positive for cathinone and cathine. The results showed that cathinone concentration was 0.03, 0.03, 0.06, 0.07, 1.85 and 31 μg/ml in brain, liver, blood, vitreous humor, stomach and chewed green plant, respectively. Whereas, the concentration of cathine was 0.31, 3.28, and 141 μg/ml in kidney, stomach and chewed green plant, respectively. Cathine and cathinone concentrations were found to be changed with respect to site of sampling. The results suggest that stomach and chewed green plants are considered as good samples to show the concentration for both cathine and cathinone at the time of death of the khat chewer.

Khat (Catha edulis Forsk) is a natural psychoactive substance which contains two main addictive substances; Cathine and Cathinone. Khat is widely used in east Africa and southern Arabian Peninsula. Khat chewers believe that it improves work capacity and increases energy level and alertness. That is why we aimed in this study to investigate the physiological and ergogenic effects of khat extract.This study is an experimental study conducted at the Substance Abuse Research Centre in Jazan University, Saudi Arabia. Thirty healthy young volunteers were randomly assigned into two experimental groups. The first group ingested 45 g of grounded khat leaves extract mixed with juice in the first session then placebo (juice only) in the second session. While the second group ingested the placebo in the first session and the grounded khat leaves with juice in the second session. Experiments were done between December 2012 and March 2013. We recorded the blood pressure, heart rate, grip strength, and reaction time every 15 min for 75 min after each ingestion. The study proposal was reviewed and approved by Research Ethics Committee (REC) of the Medical Research Centre in Jazan University.The results showed the consumption of 45 g of grounded khat leaves contributed to the increase in blood pressure (SBPDBP) and reaction time (p0.05); but had no significant effect on heart rate and grip strength (p0.05).The findings of this study showed that Khat has an acute effect on some physiological parameters. These findings support the prohibition of cathinone and cathine by the World Anti-Doping Agency (WADA, 2016).

Imaging Mass Spectroscopy (IMS) is a unique research tool providing localization and identification of a wide range of biomolecules as essential data to understand biochemical processes in living organisms. Secondary Ion Mass Spectrometry with high-energy heavy ions (MeV-SIMS) is emerging as a promising IMS technique for chemical imaging of biological tissue.We measured the molecular mass spatial distributions in leaves of khat (Catha edulis). Khat is a natural drug plant, native to eastern Africa and the Arabian Peninsula. In these countries, fresh leaves are being chewed by significant part of population. It was reported that 80% of the adult population in Yemen chew the khat leaves. The main stimulating effects of khat are induced by a monoamine alkaloid called cathinone. During leaf ageing, cathinone is further metabolised to cathine and norephedrine. Earlier studies identified the alkaloids in khat, however little is known on their spatial distribution, reflecting the biosynthesis and accumulation in the tissue.Chemical mapping of alkaloids on cross-sections of khat leaves by MeV-SIMS was done at JSI by a pulsed 5.8 MeV Cl-35(6+) beam, focused to a diameter of 15 mu m, using a linear time-of-flight (TOF) spectrometer with a mass resolution of 500. In addition, measurements of MeV-SIMS mass spectra were performed at Kyoto University by a continuous broad beam of 6 MeV Cu-63(4+) ions at an orthogonal TOF spectrometer with a high mass resolution of 11,000. Sections of leaves were analysed and mass spectra obtained at both MeV-SIMS setups were compared. Tissue-level distributions of detected alkaloids are presented and discussed. (C) 2017 Elsevier B.V. All rights reserved.

Background: Catha edulis Forsk. (Khat) is traditionally used for treating various disorders. Nevertheless there are no reports of any scientific assessment of its psychopharmacological properties. Objective: Therefore, the current study was designed to evaluate the antidepressant-like activity of Khat ethanolic extract using established animal models of depression and stress. Materials and Methods: Ninety healthy male albino mice were used in this study. Forced swim, tail suspension and head poking tests were utilized to evaluate the antidepressant-like activity of the ethanolic extract of Khat (100, 200 and 400 mg/kg, i.p.) and escitalopram (standard drug) which were administered 30 min prior to the tests. Phytochemical analysis of the standardized extract was conducted using liquid chromatography-mass spectroscopy (LC-MS). Results: A significant decrease in the head-dipping behavior was noticed after administration of 100, 200 and 400 mg/kg of Khat extract. Moreover, the extract significantly decreased the immobility time in tail suspension and forced swim tests. The presence of cathinone and cathine were detected in the extract using LC-MS. Conclusion: The current results suggest that the extract of Khat leaves has acute antidepressant properties and may have sedative effects. SUMMARY Antidepressant-like activity of Khat established in vivoThe extract decreased the immobility time in tail suspension and forced swim testsLiquid chromatography-mass spectroscopy data revealed the presence of cathinone and cathine in Khat extract. Abbreviations used: LC-MS: Liquid chromatography-mass spectroscopy; NIST: National institute of standard technology; SSRI: Serotonin reuptake inhibitors; FST: Forced swim test; TST: Tail suspension test.

Introduction Catha edulis (qat, khat, mirra) is a woody plant species that is grown and consumed in East Africa and Yemen for its stimulant alkaloids cathinone, cathine and norephedrine. Two Celastraceae species, in addition to qat, have been noted for their stimulant properties in ethnobotanical literature. Recent phylogenetic reconstructions place four genera in a clade sister to Catha edulis, and these genera are primary candidates to search for cathine and related alkaloids. Objective Determine if cathine or related alkaloids are present in species of Celastraceae other than Catha edulis. Methods Leaf samples from 43 Celastraceae species were extracted in water followed by basification of the aqueous extract and partitioning with methyl-t-butyl ether to provide an alkaloid-enriched fraction. The extract was derivatised in a two-stage process and analysed using GC–MS for the presence of cathine. Related alkaloids and other metabolites in this alkaloid-enriched fraction were tentatively identified. Results Cathinone, cathine and norephedrine were not detected in any of the 43 Celastraceae species assayed other than Catha edulis. However, the phenylalanine- or tyrosine-derived alkaloid phenylethylamine was identified in five species. Nine species were found to be enriched for numerous sterol- and terpene-like compounds. Conclusion These results indicate that cathine is unique to Catha edulis, and not the compound responsible for the stimulant properties reported in related Celastraceae species. Copyright © 2017 John Wiley & Sons, Ltd.

Objective: To investigate the efficacy and safety of increasing doses of cathine (nor-pseudoephedrine) as a weight-lowering agent in patients with obesity. Methods: Overweight and obese patients (n = 241, mean BMI 34.6 ± 3.4 kg/m²) were randomly allocated to one of three doses of cathine (16 mg, 32 mg, 53.3 mg) or placebo in addition to a multimodal lifestyle intervention program in a multicenter, double-blind, controlled, dose-finding study for 24 weeks. Primary outcome was weight loss. Results: Treatment with the 3 doses of cathine resulted in a significantly greater weight loss compared to placebo over 24 weeks: 6.5 ± 4.2 kg for 16 mg cathine, 6.2 ± 4.7 kg for 32 mg cathine, and 9.1 ± 5.4 kg for 53.3 mg cathine versus 2.4 ± 4.4 kg for placebo (each p < 0.01, ANCOVA). The percentage of patients losing > 5% / >10% of initial body weight was significantly greater for all doses of cathine than for placebo (each p < 0.01, chi-square test). Heart rate increased dose-dependently (by 1.2 bpm under 16 mg, 5.8 bpm under 32 mg, and 6.2 bpm under 53.3 mg cathine), but no suspected unexpected serious adverse reactions were noted. The overall dropout rate was 24.9%, with the highest rate in the placebo group (42.3%). Conclusion: Cathine appears to be an effective weight-lowering agent for adjunct treatment of obesity, but additional clinical studies on its efficacy and safety are required.

Toxicology screening of unusual poisons is important. We report a case of death of a young female suspected to die due to drug overdose. Medico legal autopsy was indicative of cerebral hemorrhage. The deceased had no previous history of drug dependence. Toxicological analysis revealed the presence of cathine {(+)-norpseudoephedrine} and alcohol in the blood. The concentration of ethyl alcohol in blood was determined as 85.57 mg per dL. An overview of toxicology of cathine and its side effects are presented. The reports due to fatal intoxication of cathine in combination with alcohol are rarely reported. The present case substantiates that an intake of low potent drug like cathine in combination with alcohol can cause fatal intoxication.

Ethnopharmacological relevance Khat, the leaves of Catha edulis, is used as a “natural amphetamine-like” stimulant in eastern and southern Africa, as well as in the Arabian Peninsula. Leaves are masticated to elicit a state of euphoria. Although the psychostimulatory effects of the leaves are attributed to the presence of phenylpropylamino alkaloids ( i.e. cathinone, cathine and norephedrine), the extent of permeation of these alkaloids across the oral and intestinal mucosa has not been established. Materials and methods Cathinone was isolated in the form of the oxalate salt from young buds, following acid-base extraction. High performance countercurrent chromatography (HPCCC) was used to isolate cathine and norephedrine, following borohydride reduction of a mixture of the three alkaloids. The in vitro permeability of these three alkaloids in their pure form, as well as in a crude extract, was evaluated across Caco-2 cell monolayers and across excised porcine intestinal, sublingual and buccal tissues. Results The purities of the isolated cathine and norephedrine were in excess of 90%, thereby proving that HPCCC can be applied for efficient separation of these alkaloids from extracts of Khat. The apparent permeability (P app ) coefficients for the Khat alkaloids in their pure form were all above 1.0×10 −6 cm/s, indicating that the transport of the three alkaloids across the selected biological membranes is comparable to that of the highly permeable reference compound, caffeine. Although readily transported across the various membranes, the alkaloids were transported to a lesser extent when present in a leaf extract, suggesting that other phytochemicals present in the extract influence their permeation. Conclusions These results provide evidence that chewing of Khat contributes to the buccal and sublingual absorption of the psychoactive alkaloids in the bloodstream directly across the oral mucosal membranes. In addition, it confirms that these metabolites will be readily absorbed from the gastrointestinal tract when swallowed.

© 2019. AJFSFM. This is an open access article, distributed under the terms of the Creative Commons, Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/).

The stimulating herbal drug khat (cathine, cathinone) and its analog methcathinone are common substances of abuse in most countries. A GC-MS method was developed and validated for the detection and quantification of cathine, cathinone, methcathinone and ephedrine in oral fluid specimens. The analytes and internal standard (amphetamine-d5) were extracted from 0.5 mL oral fluids by ethyl acetate, and then the dried extracts were derivatized with heptafluorobutyric anhydride at 70°C for 30 min. The MS was used in selected ion monitoring mode. Ions monitored were m/z 117, 240 and 330 for cathine, m/z 77, 105 and 240 for cathinone, m/z 105, 210 and 254 for methcathinone, m/z 210, 254 and 344 for ephedrine and m/z 244 and 336 for amphetamine-d5 The calibration curves were linear (r(2)> 0.98) in the concentration range 20-2,000 ng/mL for all analytes. The intra- and inter-assay imprecisions were within (1.6-12.5%) and (1.5-9.5%), respectively, for all analytes. Intra-assay accuracies were between -5.9 and 6.7% for all analytes. The method was successfully applied to detect and quantify the target analytes from oral fluid specimens collected from Khat and methcathinone users.

Despite the well-established medicinal values of Jimson weed (Datura stramonium L.), this medicinal plant has been associated with neurological effects such as hallucination and anxiety in folklore. This study examined the effect of alkaloid extracts from the leaf and fruit of Jimson weed on critical enzymes of the monoaminergic [monoamine oxidase (MAO)] and cholinergic [acetylcholinesterase (AChE) and butyrylcholinesterase (BChE)] systems of neurotransmission. Alkaloid extracts were prepared by solvent extraction method and their interaction with the activities of MAO, AChE, and BChE were assessed (in vitro). Gas chromatography coupled with mass spectroscopic (GC-MS) characterization of the extracts was also carried out. The results revealed that the extracts inhibited the activity of the enzymes assayed for in a concentration-dependent manner. Considering the IC50 values, the fruit extract had more potent (P

A literature criticism is given on methods using currently gas chromatography mass spectrometry (GC/MS) to determine cathine (CAT), cathinone (CTN) and norephedrine (NE), jointly khatamines. In this study, khatamines' oximation, trimethylsilylation and mass fragmentation properties-applying N-Methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA), its trimethyliodosilane (TMIS) catalyst containing version (MSTFA(TMIS)), N,O-bis(trimethylsilyl)trifluoroacetamide (BSTFA) and hexamethyldisilazane (HMDS)-was highlighted, at first. Derivatization, mass fragmentation and quantitation related, optimized model investigations have been carried out as a function of the reaction times and conditions. Special emphasis was put (i) on the stability of the primarily formed (CAT-2TMS, NE-2TMS, CTN-TMS(TMS-oximes)1,2), then transformed, fully derived (CAT-3TMS, NE-3MTS, CTN-2TMS(TMS-oximes)1,2) species, and, (ii) on the proportionally formed stable products, suitable to selective quantitation of all three natural amines, simultaneously. Results, as novelty to the field confirmed that (i) TMIS catalyzed trimethylsilyation triggers to form fully derivatized species unfortunately, in part only; while, (ii) khatamines' simultaneous quantitation needs to be carried out in a two steps derivatization process consisting of oximation (1st step, hydroxylamine in pyridine) and trimethylsilylation (2nd step, MSTFA), to the CAT-2TMS, NE-2TMS, CTN-TMS(TMS-oximes)1,2. These species were characterized with their retention, mass fragmentation and analytical performance properties, in model solutions and in the presence of plant tissues, as well: R(2), limit of quantitation (LOQ) data, expressed in pg/1μL injection basis, proved to be 62.5pg (CAT), 20pg (NE) and 62.5pg (CTN), respectively. The practical utility of proposal was enormously enhanced by the novel, direct sample preparation method. In this process, the freshly harvested, freeze-dried, then pulverized leaves of Catha edulis FORKS were directly derivatized, in the presence of the matrix. Reproducibility (in average 2.07 RSD% varying between 0.15 and 5.5 RSD%), linearity (0.9990-0.9994) and recovery (95.7-99.1%) values of the new sample preparation protocol was confirmed by the standard addition method for CAT, NE and CTN equally. From plant leaf, 0.061w/w% CAT and 0.014w/w% NE contents were obtained. In this tissue CTN was not found. Very likely attributable to the unfavorable climate for the plant: grown in Hungary of temperate zone and naturalized in the tropical Africa.

ObjectiveKhat is a plant of the Celastraceae family that is chewed for several hours/day in Yemen and most of the East African countries. Cathinone and cathine are the main Khat components and are structurally and functionally related to amphetamine. The present study has been designed to assess levels of antioxidants in the saliva of Khat chewers.MethodsSaliva samples of 50 volunteers were collected from Khat-chewers and non Khat-chewers, 25 samples each. Saliva samples were collected and used for measurments of salivary antioxidant system including; catalase, total and protein thiols, glutathione and uric acid (UA). Moreover, activity of α-amylase and lactate dehydrogenase (LDH) and levels of total protein, glucose, and cholesterol were also measured.ResultsThe activities and levels of antioxidants were significantly decreased in Khat-chewing group as compared to controls (p

A molecularly imprinted polymer (MIP) was prepared using (-)-norephedrine as the template, methacrylic acid as the functional monomer, ethylene glycol dimethacrylate as the cross-linker and chloroform as the porogen. The MIP was used as a selective sorbent in the molecularly imprinted solid-phase extraction (MIP-SPE) of the psychoactive phenylpropylamino alkaloids, norephedrine and its analogs, cathinone and cathine, from Khat (Catha edulis Vahl. Endl.) leaf extracts prior to HPLC-DAD analysis. The MIP was able to selectively extract the alkaloids from the aqueous extracts of Khat. Loading, washing and elution of the alkaloids bound to the MIP were evaluated under different conditions. The clean baseline of the Khat extract obtained after MIP-SPE confirmed that a selective and efficient sample clean-up was achieved. Good recoveries (90.0-107%) and precision (RSDs 2.3-3.2%) were obtained in the validation of the MIP-SPE-HPLC procedure. The content of the three alkaloids in Khat samples determined after treatment with MIP-SPE and a commercial Isolute C18 (EC) SPE cartridge were in good agreement. These findings indicate that MIP-SPE is a reliable method that can be used for sample pre-treatment for the determination of Khat alkaloids in plant extracts or similar matrices and could be applicable in pharmaceutical, forensic and biomedical laboratories. Copyright © 2015 John Wiley & Sons, Ltd.

The use of khat (Catha edulis) while on medication may alter treatment outcome. In particular, the influence of khat on the metabolic activities of drug-metabolizing enzymes is not known. We performed a comparative 1-way crossover study to evaluate the effect of khat on cytochrome P450 (CYP)2D6 and CYP3A4 enzyme activity. After 1 week of khat abstinence, baseline CYP2D6 and CYP3A4 metabolic activities were determined in 40 Ethiopian male volunteers using 30 mg dextromethorphan (DM) as a probe drug and then repeated after 1 week of daily use of 400 g fresh khat leaves. Urinary concentrations of cathinone and cathine were determined to monitor the subjects' compliance to the study protocol. Genotyping for CYP2D6*3 and CYP2D6*4 was done. Plasma DM, dextrorphan and 3-methoxymorphinan concentrations were quantified. CYP2D6 and CYP3A4 enzyme activities were assessed by comparing plasma log DM/dextrorphan and log DM/methoxymorphinan metabolic ratio (MR) respectively in the presence and absence of khat. Cytochrome 2D6 MR was significantly increased from baseline by concurrent khat use (paired t test, P = 0.003; geometric mean ratio, 1.38; 95% confidence interval [95% CI], 1.12-1.53). Moreover, the inhibition of CYP2D6 activity by khat was more pronounced in CYP2D6*1/*1 compared with CYP2D6*1/*4 genotypes (P = 0.01). A marginal inhibition of CYP3A4 activity in the presence of khat was observed (P = 0.24). The mean percentage increase of CYP2D6 and CYP3A4 MR from baseline by khat use was 46% (95% CI, 20-72) and 31% (95% CI, 8-54), respectively. This is the first report linking khat use with significant inhibition of CYP2D6 metabolic activity in humans.

Ephedrines are widely used in therapy. Because of their stimulant properties, these substances are relevant in different forensic fields. At present, the state of the art for ephedrines quantification relay based on a liquid chromatography mass spectrometry, mainly because of the dilute-and-shoot approach. Notwithstanding, several gas chromatography based methods have already been described, all of them include cleanup steps, with the potential disadvantage of incurring errors and increasing the workload. In this paper, a straightforward method for ephedrine quantification based on gas chromatographic mass spectrometry, without cleanup and based on Doehlert matrix optimization is presented. Only 10 µL of a urine sample is necessary and for N-methyl-N-(trimethylsilyl)trifluoroacetamide/N-methyl-bis-trifluoracetamide derivatives, the intermediate precision was 2.77% for ephedrine, 9.20% for cathine, 8.29% for norephedrine and 4.27% for pseudoephedrine. The limit of detection was 20 ng mL-1 for ephedrine, 30 ng mL-1 for cathine and 40 ng mL-1 for norephedrine and pseudoephedrine.

Khat (or Catha edulis) is an evergreen tree of the Celastracea family. It was originated from Ethiopia, but was later distributed to different countries. This plant is a natural stimulant which contains two main substances: Cathine and Cathinone that caused excitement, euphoria, and loss of appetite. Although khat has some benefits, it harms the body and caused many other severe effects such as addiction and illness. It also affects the social and economic growth negatively. Some studies are proved that Khat chewers may become psychologically addicted to khat. Hence, the World Health Organization (WHO), in 1980, categorized the Khat plant as a drug of abuse that can create mild to moderate psychological dependence. Based on WHO is classification, many countries have controlled the use of Khat by making its use illegal. Although Saudi Arabia is one of the countries that forbid Khat use, the majority of the population, southern region, is accepting and using it easily. However, this research clarifies the prevalence of Khat use among students in the Southern region of Saudi Arabia, and concludes the different factors which contribute to chewing Khat among students.

Recently, a class of new psychoactive substances (NPS) has appeared as drugs of abuse. NPS comprise different drug classes; the most popular are synthetic cannabinomimetics and designer analogues of cathinone, an intermediate metabolite in a biosynthetic pathway, and a psychostimulant, found in the plants Catha edulis Forsk., Ephedra gerardiana sikkimensis, and Ephedra sinica. In C. edulis and E. gerardiana sikkimensis, cathinone is converted to norpseudoephedrine (cathine) and norephedrine, while in E. sinica the pathway continues to pseudoephedrine and ephedrine. While cathinone analogues are used for recreational purposes, cathinone, itself, is consumed only through chewing khat, i.e., the fresh leaves and twigs of the plant C. edulis Forsk., a practice similar to chewing of coca leaves. The use of khat is common among people living in Southwestern Arabia and Eastern Africa with an estimated 10–20 million daily users. This chapter describes the use of khat, its distribution and cultivation, the biosynthetic pathway that involves cathinone, the long delay in the discovery of cathinone, the neurochemistry of khat and cathinone, its effects on humans, and its dependency and addiction potential.

Khat (Catha edulis) belongs to Celastraceae family which contains 60–70 genera and 850–900 species. It is an indigenous plant to Ethiopia and Yemen as the countries of origin. It is also found in many other east and southern African countries. Khat leaves are chewed by the local people for their stimulant action. The main active ingredient compounds those are responsible for this action is cathinone and a mild stimulant cathine. In addition to these khat contains several phytochemicals such as alkaloids (phenylalkylamines and cathedulins), flavonoids, steroid and triterpenoids, monoterpenes and volatile aromatic compounds, and other miscellaneous compounds like vitamins and amino acids. Hence, this paper presents a comprehensive and unified review of literatures which concerned on the phytochemical composition of khat plant. And it also provides the isolated compounds with their chemical structures.

Consumption of Ephedra alkaloids has been prohibited in-competition by the World Anti-Doping Agency (WADA). In Taiwan, colds are often treated with Chinese herbal formulae containing Herba Ephedrae. We screened the products sold in Taiwan and preliminarily assessed their relationships with WADA threshold violations. Fifty-six concentrated powder products, including 19 Chinese herbal formulae that contained Herba Ephedrae, were collected. The content of Ephedra alkaloids, namely ephedrine (E), methylephedrine (ME), norpseudoephedrine (NPE; cathine), pseudoephedrine (PE), and norephedrine (NE; phenylpropanolamine), was determined using a validated high-performance liquid chromatography method. The results revealed that the phenotypic indicators of the collected products, E/PE and E/total ratios, were 1.52–4.70 and 0.49–0.72, respectively, indicating that the Herba Ephedrae species in these products was probably E. sinica or E. equisetina, but not E. intermedia. The contents of E, ME, NPE, PE, and NE and the total alkaloid contents in the daily doses of the products were 0.45–34.97, 0.05–4.87, 0.04–3.61, 0.15–12.09, and 0.01–2.00 mg and 0.68–53.64 mg, respectively. The alkaloid contents followed a relatively consistent order (E > PE > ME ≈ NPE > NE), even for products from different manufacturers. We calculated that single doses of 50.0% and 3.6% of the products would result in the WADA thresholds of E and NPE being exceeded, respectively. Our data provide critical information for athletes and medical personnel, who should be wary of using complex Chinese herbal formulae in addition to over-the-counter products.

A broad forensic screening method for 256 analytes in whole blood based on a fully automated SPE robotic extraction and ultra-high-performance liquid chromatography (UHPLC) with TOF-MS with data-independent acquisition has been developed. The limit of identification was evaluated for all 256 compounds and 95 of these compounds were validated with regard to matrix effects, extraction recovery, and process efficiency. The limit of identification ranged from 0.001 to 0.1 mg/kg, and the process efficiency exceeded 50% for 73 of the 95 analytes. As an example of application, 1335 forensic traffic cases were analyzed with the presented screening method. Of these, 992 cases (74%) were positive for one or more traffic-relevant drugs above the Danish legal limits. Commonly abused drugs such as amphetamine, cocaine, and frequent types of benzodiazepines were the major findings. Nineteen less frequently encountered drugs were detected e.g. buprenorphine, butylone, cathine, fentanyl, lysergic acid diethylamide, m-chlorophenylpiperazine, 3,4-methylenedioxypyrovalerone, mephedrone, 4-methylamphetamine, p-fluoroamphetamine, and p-methoxy-N-methylamphetamine. In conclusion, using UHPLC-TOF-MS screening with data-independent acquisition resulted in the detection of common drugs of abuse as well as new designer drugs and more rarely occurring drugs. Thus, TOF-MS screening of blood samples constitutes a practical way for screening traffic cases, with the exception of δ-9-tetrahydrocannabinol, which should be handled in a separate method.

The analysis of sewage for the residues of commonly used illicit drugs has successfully been applied as a suitable approach for estimating community illicit drug use. The drug market is increasingly dynamic with new substances continually being marketed for recreational purposes. In this study, ultra-high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC-MS/MS) was used to simultaneously and quantitatively detect the exogenous biomarkers of new classes of recreational drugs in sewage collected from three different Norwegian cities (Oslo, Bergen, Hamar). The samples were screened for the presence of khat (d-norpseudoephedrine and cathinone), mephedrone, pseudoephedrine, 7-aminoflunitrazepam, para-methoxyamphetamine (PMA), para-methoxy-N-methylamphetamine (PMMA) and a selection of urinary metabolites of synthetic cannabinoids collectively termed ´Spice´ (5-3-1-naphthoyl-1H-indol-1-yl-pentanoic acid (JWH 018 N-pentanoic acid), 1-5-hydroxypentyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 018 N-5-hydroxypentyl), 4-3-1-naphthoyl-1H-indol-1-yl-butanoic acid (JWH 073 N-butanoic acid), 1-4-hydroxybutyl-1H-indol-3-ylnaphthalen-1-yl-methanone (JWH 073 N-4-hydroxybutyl), 1-5-hydroxypentyl-1H-indol-3-yl4-methylnaphthalen-1-yl-methanone (JWH 122 N-5-hydroxypentyl), 1-5-fluoro-4-hydroxypentyl-1H-indol-3-ylnaphthalen-1-ylmethanone (AM2201 N-4-hydroxypentyl), and 1-5-hydroxypentyl-1H-indol-3-yl4-methoxyphenyl-methanone (RCS-4 N-5-hydroxypentyl)). Limits of detection were 1 ng/L for amphetamine like compounds and 5 ng/L for the metabolites of synthetic cannabinoids while the limits of quantification were 3 and 15 ng/L, respectively. Three of the fourteen selected biomarkers (cathine, pseudoephedrine and the synthetic cannabinoid metabolite JWH-018 N-5-hydroxypentyl) were detected in sewage, alongside the illicit drugs (and/or metabolites) typically found in sewage (cocaine, benzoylecognine, methamphetamine, MDMA, and THC-COOH). The khat biomarker d-norpseudoephedrine was detected in Oslo sewage at a mean concentration of 93 ng/L that represents a daily load of 54 mg/day/1000 inhabitants. Pseudoephedrine was present at mean concentrations of between 27 and 67 ng/L representing normalized daily loads of between 10 (Hamar) and 24 mg/day/1000 inhabitants (Bergen). The daily normalized loads of JWH-018 N-5-hydroxypentyl were between 49 (Oslo) and 62 mg/day/1000 inhabitants (Hamar). This study demonstrates for the first time that sewage biomarker analysis can be applied to evaluate not only the use the traditional illicit drugs (cocaine, cannabis and amphetamines), but also the use of certain new synthetic drugs.

Many studies have found that some dietary supplement product labels do not accurately reflect the actual ingredients. However, studies have not been performed to determine if ingredients in the same dietary supplement product vary over time. The objective of this study was to assess the consistency of stimulant ingredients in popular sports supplements sold in the United States over a 9-month period. Three samples of nine popular sports supplements were purchased over the 9-month period. The 27 samples were analyzed for caffeine and several other stimulants (including adulterants). The identity and quantity of stimulants were compared with stimulants listed on the label and stimulants found at earlier time points to determine the variability in individual products over the 9-month period. The primary outcome measure was the variability of stimulant amounts in the products examined. Many supplements did not contain the same number and quantity of stimulants at all time points over the 9-month period. Caffeine content varied widely in five of the six caffeinated supplements compared with the initial measurement (–7% to +266%). In addition, the stimulants—synephrine, octopamine, cathine, ephedrine, pseudoephedrine, strychnine, and methylephedrine—occurred in variable amounts in eight of the nine products. The significance of these findings is uncertain: the sample size was insufficient to support statistical analysis. In our sample of nine popular sports supplements, the presence and quantity of stimulants varied over a 9-month period. However, future studies are warranted to determine if the variability found is significant and generalizable to other supplements.

Background:\ud Khat (Catha Edulis Forsk) is a natural psychoactive substance that contains addictive substances such as Cathine and Cathinone which have similar structure and action to amphetamine. This substance has been suggested that it can decrease perceived exertion and thus improve performance. There is no study in the literature regarding the effect of khat on exercise performance. Therefore, the aim of this study is to find out whether khat leaves can decrease perceived exertion in humans.\ud \ud Methods:\ud This study is an experimental crossover study conducted at the Substance Abuse Research Centre in Jazan University, Saudi Arabia. Twenty one healthy volunteers were randomly assigned into two experiment trials. Each volunteer visited the lab three times. The first visit was a familiarization session about the nature of the study and the equipment. On the second visit, 45 min before the experiment volunteers ingested either 33 ml of fruit juice (placebo) or the juice mixed with 45 g of ground khat leaves. Then the participants were instructed to perform a 10 Km cycling on an ergometer and recorded the following physiological variables repeatedly on every 5 min of cycling: heart rate, time to complete 10 km cycling, tympanic temperature, and perceived exertion rate. On the third visit a crossover trial was conducted one week after the second visit; then the same cycling test was performed and the same variables were recorded as the second visit. The experimental protocol was reviewed and approved by Research Ethical Committee of the Medical Research Centre, Jazan University.\ud \ud Results:\ud According to study results, khat dramatically decreased time taken to complete a 10 km cycling time trail (p

A simple and accurate liquid chromatography/tandem mass spectrometry (LC/MS/MS) method has been developed and validated for the quantitative determination of ephedrine, pseudoephedrine, methylephedrine, cathine, salbutamol, morphine and epitestosterone in human urine. Urine samples were spiked with internal standard and diluted with acetonitrile. After centrifugation, the supernatants were directly analyzed by LC/MS/MS using the selected reaction monitoring (SRM) mode. The linearity, intra- and inter-day precision, accuracy, limit of detection (LOD) and limit of quantification (LOQ) were evaluated and the method was found to be accurate and reproducible for the quantitation of threshold substances. When the method was applied to the analysis of blind urine samples for the proficiency test, the results were close to the nominal concentrations, within 87.7-106.6% of nominal values, suggesting that the developed methods can be successfully applied to routine doping analyses.

A liquid-chromatography-tandem-mass-spectrometry method using pneumatically assisted electrospray ionisation (LC-ESI-MS/MS) was developed for the simultaneous determination of cathinone, methcathinone, ethcathinone, amfepramone, mephedrone, flephedrone, methedrone, methylone, butylone, cathine, norephedrine, ephedrine, pseudoephedrine, methylephedrine and methylpseudoephedrine in human live and post-mortem whole blood. The blood proteins were precipitated by the addition of methanol, and the extract was purified by ultrafiltration. The separation of diastereomeric ephedrines was achieved on an ethyl-linked phenyl column. Matrix-matched calibrants combined with the isotope dilution of selected substances were used for quantitative analysis. The relative intra-laboratory reproducibility standard deviations were generally better than 7% at concentrations of 20 μg/L, and the mean true recoveries were 87-106% in the concentration range of 10-250 μg/L. The detection limits were in the range of 0.5-3 μg/L. The cathinones were unstable in whole blood and sample extracts under neutral conditions, but the stability could be improved by the acidification of the sample matrix.

A primary concern with the forensic analysis of the khat plant (Catha edulis) has been the need to preserve the principle psychoactive component, cathinone, which converts to the less-active substance, cathine, after harvesting. The loss of cathinone has serious legal implications since it is a Schedule I controlled substance under federal regulations in the United States, while cathine is Schedule IV. A common misconception is that cathinone is highly unstable once the plant is harvested, and may be undetectable upon drying and prolonged storage. However, drying the plant material will preserve cathinone. Numerous seizures of a dried form of khat (referred to as "graba" in the United States) have been made in recent years, suggesting that drying the plant material is a viable approach to preserve khat evidence for both storage and reanalysis. A qualitative and quantitative study of the composition of khat samples seized as dried plant material has found the khat alkaloids to be relatively stable for a monitored period of 3 years, and cathinone has remained identifiable while stored at room temperature for over 10 years. Studies of green khat (received moist) have also determined that drying the moist leaves at either room temperature or by the application of heat are suitable methods to preserve cathinone in the dried material. These findings demonstrate that cathinone persists in dried khat for a time frame of several years, and simple drying techniques are an effective means to preserve seized khat evidence for long-term storage.

Background: Catha edulis is an evergreen plant belonging to the Celastraceae family, used by the people of Africa, Yemen, and some part of Saudi Arabia as a recreational plant. They chew it alone or in combination with alcohol for attaining euphoria. Objective: There are contradictory studies in the genotoxic potential of the leaves of this plant. So far, the accomplished studies were conducted with much high doses. Hence, the current study has been designed to check the genotoxicity of khat-leaves extract with subcytotoxic concentration in human peripheral blood lymphocytes. Materials and Methods: We have employed comet assay, micronuclei (MN) analysis, and Ames test to evaluate the objectives. Results: The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay has revealed that there is a dose-dependent reduction in cell viability with an IC50value of 76.5 μg/ml at 24-h treatment. The results of the comet assay have showed a nonsignificant difference between control and treatment up to 40 μg/ml extract. There was no dose-dependent increase in the frequency of binucleated cells with MN and reduction of cell proliferation in all the doses. The extract showed no mutagenic effect, in both TA98 and TA100 strains even with S9 activation. Conclusion: Under the experimental conditions employed in the present study, khat has been found to be nongenotoxic at subcytotoxic level. Abbreviations used: DMSO: Dimethyl sulfoxide; GIT: gastrointestinal tract; HPBL: Human peripheral blood lymphocytes; KCL: Potassium chloride; LCQ: Low-content quantification; MMC: Mitomycin C; RPMI: Roswell park memorial institute; SWGDRUG: Scientific working group for seized drugs.

Background: Khat (Catha edulis Forsk) is a shrub endemic to East Africa and the Arabian Peninsula that is widely consumed for its psychoactive properties. Due to the increase of illicit drugs consumption in the world, there is a need for fast investigation techniques in order to rapidly characterize the huge number of seized samples. In the recent years, direct analysis in real-time mass spectrometry (DART-MS) proved to be a useful and convenient method which allows recording mass spectra of various samples without any preparation. Objectives: The present work describes a convenient procedure for rapid characterization of fresh Khat samples by identification of cathinone and cathine which are the most active constituents in different parts of the plant. Materials and Methods: A fresh sample of C. edulis was cut into small pieces with separation of leaves and stems; each part was investigated using DART-time-of-flight (ToF)-MS. The fresh samples were submitted to a stream of hot helium as vaporization and ionization gas, without any extraction or prior treatment. Results: The obtained spectra showed many peaks corresponding to the mass range from 100 to 600 Da. Cathinone and cathine which are known to be the main psychoactive components in C. edulis were found in all parts of the fresh samples; their protonated peaks appeared at 150 and 152 Da, respectively. The peak intensity of these two characteristic alkaloids was notably different between leaves and stems; it roughly decreased from lower to upper leaves, while their highest content was obtained in the upper stem. Conclusion: The present work showed that DART-ToF-MS is a powerful tool for fast investigation of real samples, particularly in the field of forensic sciences.

Pseudoephedrine (PSE) as a sympathomimetic is an ingredient of many proprietary medicines which are available on the medical market over the counter (OTC drugs). It can be converted to cathine (CATH, norpseudoephedrine) inside the body. Until the end of 2003, PSE had been a banned substance in sport in case its urinary concentration was greater than 25 mircog/ml. Then the World Anti-Doping Agency (WADA) removed PSE from the prohibited list. Prior to 2004 CATH was a forbidden substance and it is still one. CATH is included on the WADA prohibited list in the group of stimulants. The results of a doping control concerning PSE conducted in the Department of Anti-Doping Research of Institute of Sport in Warsaw in the years 2001-2003 and 2004-2007 have been compared. Moreover, several dozen of urine samples collected from the patients taking OTC drugs with PSE have been analysed. In these samples the concentration of PSE and CATH has been estimated. The results of this study have shown that athletes were using PSE frequently and in high doses between 2004 and 2007 when this substance was permitted by WADA. It is possible that athletes can obtain a positive result of doping control with CATH after the use of PSE.

Introduction. Khat (Catha edulis) is an evergreen tree/shrub that is thought to affect sexual motivation or libido. Its positive effect on sexual desire is more frequently observed in females than in males and occurs when khat is chewed. Thus, khat’s effects on sexual behavior may depend on the release mode of its active constituent. Aim. This study aimed to investigate the effect of dried khat alkaloids on the sexual motivation and estradiol levels of female rats, with special emphasis on the importance of the sustained release effect. Methods. Dried khat leaves were extracted and isolated. The alkaloids in khat extract were identified and calculated using thin layer chromatography and high-performance liquid chromatography. The isolated khat extract was microencapsulated using a phase separation coacervation method. The morphology, particle size, yield, drug loading, and entrapment efficiency were evaluated. The in vitro release and stability of alkaloids in khat extract and in khat extract microcapsules were determined. The effect of khat extract microcapsules and varying doses of khat extract on sexual motivation in female rats were investigated. Additionally, estradiol levels, vaginal secretions and vaginal pH were determined. Main Outcome Measures. The differences in the effect of khat extract and khat extract microcapsules on sexual motivation, vaginal secretion and estradiol levels in female rats were compared. Results. Cathine and norephedrine were identified in the isolated khat extract at composition of 81.3% and 17.2%, respectively. Among the formulations studied, khat extract microcapsules of formulation 2:3:5 (containing a ratio of khat extract to ovalbumin to gelatin of 2:3:5) were found to exhibit higher yield, loading, and entrapment efficiency. Khat extract microcapsules showed sustained in vitro release and were more stable than khat extract. In addition, khat extract microcapsules enhanced sexual motivation, increased vaginal secretions, and upregulated estradiol level in female rats. Conclusion. The sustained release of alkaloids from dried khat has significantly enhanced the sexual motivation and increased the estradiol level of female rats. Thus the release of dried khat alkaloids from microcapsules might be an effective means of enhancing the libido in females. Aziz HA, Peh KK, and Tan YTF. Extraction and microencapsulation of khat: Effects on sexual motivation and estradiol level in female rats. J Sex Med 2009;6:682– 695.

Khat (Catha edulis Forsk., Celastraceae) is a perennial shrub that was introduced to Israel by Yemenite immigrants. Its young leaves are chewed for their psycho-stimulating properties. Young khat leaves contain the phenylpropylamino alkaloids (-)-cathinone [(S)-α-aminopropiophenone], (+)-cathine [(1S)(2S)-norpseudoephedrine], and (-)-norephedrine [(1R)(2S)-norephedrine] as the main active principles. A novel GC-MS analysis method for the quantitative determination of phenylpropylamino alkaloids and their putative biosynthetic precursor 1-phenylpropane-1,2- dione in khat leaves was developed. We utilized an alkaline-organic extraction, coupled with gas chromatography and a chiral permethylated beta cyclodextrin phase, to allow a full separation between the two diastereoisomers (1S)(2S)-cathine and (1R)(2 S)-norephedrine. We found a marked diversity in the phenylpropylamino alkaloid content and composition in three different locally grown accessions and the commercial cultivar ‘Mahanaim’.

Background The psychostimulant khat (Catha edulis Forsk), is a herbal drug cultivated and chewed as a recreational and socializing drug in East Africa and the Arabian Peninsula for centuries. Due to increasing air transportation and the loosening of customs restrictions, it is now readily available in the Western Countries mainly used by immigrants from khat growing areas causing a concern to policy-makers. Objective We conducted this review to further gain an insight to the neuropharmacological effects of khat. Methodology PubMed search engine with key terms ‘khat’ or ‘qat’ or ‘mirra’ or’qaad/jaad’ or ‘cathinone’ was used to obtain articles relevant to khat chewing. In total 284 English written articles published from 1959 to 2007 were screened. Results Most of the studies focused on cathinone, the postulated active psychostimulant alkaloid in khat. There were few studies which investigated the entire plant extract in either in vitro or animal studies. In the majority of the studies it was reported that both cathinone and cathine, another psychoactive constituent, have actions that are similar to those of amphetamine. Conclusions It seems that the well investigated khat alkaloids have many features similar to amphetamines; however there is a need for a more thorough examination of khat itself in well designed in vitro, animal and human studies with a range of comparator drugs before confirming the claim that khat is a “natural amphetamine”.

Leaves of khat ( Catha edulis ) are masticated to elicit their psycho-stimulating properties, resulting from the presence of the phenylpropylamino alkaloids. The determination of these alkaloids is important in pharmacological, phytochemical, forensic and law enforcement environments. In this study, the use of strong cation exchange-solid phase extraction (SCX-SPE) was investigated as an alternative means of sample purification prior to the determination of cathinone, cathine and norephedrine by reversed phase (C 18 ) high performance liquid chromatography (HPLC). Extraction parameters for SCX, including loading capacity and washing solvents, were established. An existing liquid-liquid extraction (LLE) method was improved in terms of recoveries obtained, by using ethyl acetate as extractant. For pure standards of the khat alkaloids, recoveries ranged from 83 to 97%. Preconcentration, using a Genevac evaporator after the addition of acidified water to the sample, restricted analyte losses when compared to concentration under nitrogen. Although comparable recoveries were obtained when preconcentration was achieved in the presence of acidified water by rotary evaporation, this method is not suitable for large sample numbers. Best recoveries, ranging from 94 to 102%, were obtained by SCX from spiked samples. Although the extraction efficiencies of LLE were lower (87-90%), LLE yielded less complex chromatograms, indicating a purer extract. KEY WORDS : Khat alkaloids, Solid phase extraction, Liquid-liquid extraction, HPLC, Genevac Bull. Chem. Soc. Ethiop. 2015 , 29(3), 331-339. DOI: http://dx.doi.org/10.4314/bcse.v29i3.1

Khat is a natural psychoactive substance, which has been chewed for many years in Ethiopia, East Africa, and the southern Arabian Peninsula [1-3]. With the recent globalization, khat chewing has spread with African and Arabian immigrants to various Asian [4] and European [5-9] countries, and to Australia [10-12], as well as to the United States [13]. The users of khat in these new countries are predominantly immigrants from the khat chewing countries [9]. Evidence shows that fresh khat leaves contain more than 40 chemicals [15]. However, most of the stimulant effect of khat is thought to come from the chemicals cathinone, cathine, and norephedrine [14,15]. Khat users report increased levels of energy, alertness, sensations of elation, self-esteem, enhanced imaginative ability, capacity to associate ideas when using and an increase in libido [16,17]. Khat use is also associated with increased blood pressure/hypertension, development of gastrointestinal tract problems, cytotoxic effects on liver and kidneys, and keratotic lesions at the site of using [18,19].

Khat, Catha edulis Forsk, is among the most widely used plant-based psychoactive substance in the world. Grown in Eastern Africa, Horn of Africa, and southwestern part of the Arabian Peninsula, its fresh young leaves and twigs are used daily by over 20 million people for the psychostimulatory effects it produces in the user, a practice deeply rooted in the history, tradition, and culture of the indigenous population. Once hardly known outside the regions where it is grown and used, khat use has now spread to other countries. This review will cover the, phytochemistry, pharmacokinetics of the active ingredients-cathinone, cathine, norephedrine, neurochemistry, effects on cognitive and executive functions as well as its ability to produce dependency in the user. Whether it is an innocuous cultural practice or a drug of abuse is debatable as the preclinical and clinical data needed to arrive at an authoritative conclusion is lacking.