Journal of Chromatographic Science, Vol. 32, December 1994 Cd Methcathinone and Designer Analogues: Synthesis, Stereochemical Analysis, and Analytical Properties Jack DeRuiter, Lisa Hayes, Allen Valaer*, and C. Randall Clarkt Department of Pharmacal Sciences, School of Pharmacy, Auburn University, Auburn, Alabama 36849 FIT. Nogele ‘Alabama Department of Forensic Sciences, Wire Road, Auburn, Alabama 36831 This paper describes the synthesis, stereochemical analysis, and analytical properties of methcathinone and related compounds. Methcathinone represents a new class of designer street drugs that can be prepared easily from readily available starting materials, such as the ephedrines and pseudoephedrines. The oxidation of each individual isomer of ephedrine and pseudoephedrine produces homochiral methcathinone via conservation of configuration. Thus 1R,2S-ephedrine and 15,25-pseudoephedrine yield $smethcathinone, and 15,2R-ephedrine and 1R,2R- pseudoephedrine produce R-methcathinone. The isomers of methcathinone were separated by gas chromatography as the diastereomeric amides following derivatization with S-(-)-N- (triluoroacetyNprolyl chloride (TPC). The gas chromatographic-mass spectrometric analysis of methcathinone and designer analogues showed a major chromatographic peak with a mass spectrum characteristic of the parent molecule. However, the major chromatographic peak was accompanied by a secondary, well-esolved peak that yielded a molecular ion 2 mass units less than that ofthe major peak. Deuterium labeling experiments showed this minor component to arise through the thermal oxidation of the 2,3-carbon-carbon bond of the side chain to yield the 2,3-enamine. Cathinone, methcathinone, dimethcathinone, ethcathinone, and diethylcathinone (diethylpropion) were separated by reversed- phase liquid chromatography using a phenyl bonded stationary phase and an acidic (pH 3) mobile phase. Methcathinone and ‘athinone do not interfere or cross-react in standard drug abuse screening methods based on analysis by thin-layer chromatography or immunoassay. Introduction Methcathinone (ephedrone, c-methylaminopropiophenone, “CAT’) isan amphetamine-type drug of abuse that was first dis- covered in clandestine drug samples obtained in the Upper Caren az Alabama Reece abortores, ne, Souh Hl Seek, Montgoneny ‘abana 6103 *uthort whom conepondence shouldbe asd 552 Peninsula of Michigan in the early 1990s (1). More recently, this drug has been encountered in other portions of the United States, including the state of Washington (1). Furthermore, methcathinone abuse has been reported (2) in other coun- tries for some time, particularly in the former Soviet Union where it is known as “Jeff. In an attempt to prevent the more widespread manufacture and use of methcathinone in the United States, it was recently (October 15, 1993) placed into Schedule I of the Federal Controlled Substances Act (1). Methcathinone (Chart 1) is the N-methyl analogue of the natural product cathinone (‘khat”), a psychoactive alkaloid present in the leaves of the khat shrub, Catha edulis (3,4). ‘The khat plant grows in Eastern Africa and the Arabian Penin- sula, and its leaves are distributed to countries throughout the region, including Somalia. The leaves are chewed to release cathinone, which produces stimulant-like effects similar to amphetamine. It appears that only fresh khat leaves contain the active alkaloid, thus leaves must be imported daily to provide a continuous supply of the drug. This factor has historically re- stricted the availability of the khat leaves. However, with in- creased transportation efficiency, khat leaves have achieved wider distribution to more distant countries with significant African and Arab immigrant populations. Although khat use 0 NRR’ CHs =R'=H: CATHINONE =H, R'=CHy: METHCATHINONE : DIMETHCATHINONE R'=CH,CHy: ETHCATHINONE =CH,CHy: DIETHYLPROPION Chart 1, Structure of methcathinone and related compounds.Journal of Chromatographic Science, Vol. 32, December 1994 does not appear to be a significant problem in the United States, the synthetic analogue, methcathinone, has been en- countered with greater frequency, suggesting the potential for future epidemic abuse (1). The abuse of this drug, like cathi- none, appears to result from its ability to produce stimulant ef- fects by neurochemical mechanisms similar to those of am- phetamine and methamphetamine (4,5). Methcathinone was originally investigated as a potential pharmaceutical agent to treat obesity and the symptoms of depression in the 1950s and 1960s (6). However, as a result of its severe side effect profile and high addiction potential, meth- cathinone was never marketed in the United States. Itis a syn- thetic derivative of cathinone prepared from ephedrine by di- rect oxidation with dichromate or permanganate (2,6). Ephedrine is widely marketed in the United States as an “over- the-counter” dietary agent and stimulant or energy drug (“pep pills”). Thus clandestine laboratory operators can obtain ephedrine without restriction and have developed methods to convert it to methcathinone using common chemicals such as battery acid, drain cleaner, and epsom salts (1,7). Like methamphetamine, methcathinone contains a chiral carbon at the 2-position of the side chain and therefore can exist in two enantiomeric forms. Preliminary pharmacological evidence indicates that stimulant activity resides primarily in the S-(-)-isomer of methcathinone (8). Although the litera- ture (1,6) suggests that S-(-)-methcathinone can be synthe- sized stereospecifically by oxidation of 1,2S-ephedrine, no HoH NHCHS NHCHy ¢ Won, K,0r207 une, HoH NHCHs x? x So chy # Scheme 1. Synthesis of methcathinone by oxidation of ephedtines and pseudoephedrnes. direct evidence confirming the stereochemical composition of the methcathinone products is provided in these reports. Gas chromatographic and mass spectral data have been reported for methcathinone, but there are no reports on the stability of methcathinone during these analytical procedures. In our pre- liminary studies, methcathinone decomposition was observed during attempts at gas chromatographic-mass spectrometric (GC-MS) analysis. In this report, we describe the thermal de- composition products of methcathinone and designer ana- Jogues as well as their liquid chromatographic (LC) separation and the stereochemical aspects oftheir synthesis. Experimental S-(-)-N-(Trifluoroacetyl)prolyl chloride derivatization Approximately 1 mg of each methcathinone enantiomer or 2 mg of racemic methcathinone was dissolved in 10% aqueous sodium bicarbonate. This solution was extracted with approx- imately 1 mL of chloroform, S-(-)-N-(Trifluoroacetyl) prolyl chloride (TPC) reagent (250 pL of 1.0M solution in methy- lene chloride) (Aldrich Chemical; Milwaukee, WI) was added to the chloroform solutions and allowed to react for 10 min at room temperature. The solution was washed with 0.5N NaOH to remove unreacted TPC, filtered, and placed in a 2-mL capped autosampler vial for injection into the GC-MS. Gas chromatography and mass spectrometry GC-MS analyses were performed using a Hewlett-Packard 5970B mass selective detector (Wilmington, DE). All meth- cathinone samples and methcathinone-TPC derivatives, ex- cept that shown in Figure 7, were introduced into the mass spectrometer via a GC equipped with a 12-m x 0.20-mm i.d fused-silica column with a 0.33-pm film thickness of methyl- silicone (HP-1, Hewlett-Packard). For the methcathinone~TPC derivatives, the column temperature was held at 70°C for 1 min and programmed to 200°C at arate of 7.5°C/min and from 200 to 275°C at a rate of 10°C/min. For the underivatized meth- cathinone samples (except Figure 7), the column temperature was held at 70°C for 2 min and programmed to 170°C ata rate of 10°C/min and from 170 to 275°C at a rate of 25°C/min with ahold time of 2 min. The injector port temperature was 175°C. ° NHCHS W Yow, ° on™ chy # d Aer, (s-TP¢) (Pa. och i W Yew, 0 oer och A W : ZS MS ers | min ‘Scheme 2, Derivatization of methcathinone enantiomers with $--Ns(tifluoroacetylprolyl chloride The sample shown in Figure 7 was intro- duced via a GC equipped with a 10-m x 0.20-mm i.d, fused-silica column with a + | 0.33-ym film thickness of 5% cross-linked phenylmethylsilicone gum phase (DB-5, J&W; Folsom, CA). The column tempera- ture was held at 40°C for 1 min and then programmed to 150°C at a rate of 15°C/min and from 150 to 250°C at a rate of 25°C/ Liquid chromatography The liquid chromatograph (LC) consisted of a Laboratory Data Control Constametric 3000 pump (Rivera Beach, FL), 3100 Spec- 553Journal of Chromatographic Science, Vol. 32, December 1994 ‘Abundance ‘4000000 A 3000000 2000000 1000000 12 « 16 18 20 22 26 Tina (in) Abundance 000000 3000000 2000000 1000000 12 1 16 18 20 2 2 Time in) Abundance 20,720 min 166 c eoo000 700000 00000 500000 400000 300000 200000 251 7 105 100000 ul 148 ° 223 a 356 100 200 300 Auras ‘s000000 D 3000000 2000000 1000000 2 1" 16 8 20 2 28 Tine (rin) Figure 1. Gas chromatographic and mass spectral analysis of methcathinone-TPC derivatives: A chromatogram of 1R,2S-ephecrne; B, chromatogram of 15,2R- ephedrine; C, mass spectrum of methcathinone-TPC; D, chromatogram from racemic ephedrine. 554