Detection and disposition of JWH-018 and JWH-073 in mice after exposure to “Magic Gold” smoke☆
Introduction
Since the mid-2000s, various herbal incense products (HIPs) became available via the internet and at various retail outlets, initially in Europe, then more recently in the United States and Japan [1]. HIPs purport to be blends of plant materials and incense and are labeled “not for human consumption” however, they are allegedly capable of producing a marijuana-like high when smoked [1]. HIPs are usually packed in colorful foil packets containing 3–5 g of plant material. DNA testing has shown that the plant species listed as ingredients were not always present [8]. HIPS are skillfully marketed under numerous, and an ever increasing number of attractive or exotic names such as Spice Gold, Magic Gold, K-2, Summit, Lion's Tail, Buzz, Pulse, Chill Out and Yucatan Fire. Chemical analyses performed on HIPs have identified a variety of synthesized cannabimimetic compounds present [1], [2], [3], [4], [5], [6], [7], [8]. Many of these compounds interact with the CB1 receptors in the nervous system in a similar manner to Δ9-tetrahydrocannibinol (THC), the main active compound in marijuana [9], [10], [11], [12], [13]. Cannabimimetics were initially synthesized as pharmacological tools to help elucidate the structure of the cannabinoid receptor(s) in the brain and other tissues. They have also been used to assist in the search for medical applications of cannabinoids [14]. The most commonly encountered drugs are the cyclohexylphenol compounds such as CP 47, 497 [15] and naphthoylindole derivatives synthesized by John W. Huffman such as JWH-018, JWH-073 and JWH-398, Fig. 1 [16].
JWH-018 has been detected recently in the serum of two subjects after smoking 100 mg (female subject) and 150 mg (male subject) of the HIP “Smoke” which contained 2.9% JWH-018 [17]. Serum concentration were 8 and 10 ng/mL within 5 min of smoking, respectively; however, they dropped below the assay's lower limit of quantitation (LLOQ) of 0.5 ng/mL after 6 h. Another recent study of serum specimens from 80 subjects provided by various clinical and forensic sources detected JWH-018 in 9 samples at concentrations ranging from 0.30 to 8.1 ng/mL with a mean of 1.84 ng/mL [18]. JWH-073 was found in 6 samples ranging from 0.23 to 0.6 ng/mL, mean 0.42 ng/mL. Also the metabolites of JWH-018 have recently been identified in the urine of three people arrested for public intoxication [19]. Recently, Grigoryev et al. found that the major urinary metabolites of JWH-018 in man and mice are monhydroxylated metabolites and dealkylated with monhydroxylated metabolites, respectively [20]. However, there is little pharmacological or toxicological data concerning the disposition of the cannabimimetics in animals or man. To gain a better understanding of the pharmacokinetic phase of cannabimimetic intoxication, we studied the disposition of JWH-018 and JWH-073 in blood and brain of mice after controlled exposure to smoke from the HIP “Magic Gold.” These compounds were identified and quantified in whole blood and brain by high pressure liquid chromatography with ion spray positive ion triple quadruple mass spectrometry (HPLC/MS/MS). The validation of the analytical method is also presented.
Section snippets
Animals and procedures
Two sets of five C57BLC/6J mice (Jackson Laboratory, Bar Harbor, ME) were housed in the animal care quarters and maintained at 22 ± 2 C on a 12 h light/dark cycle with food and water available ad libitum. They were brought to the test environment and allowed 24 h to recover from the move. The animal study was approved by the Institutional Animal Care and Use Committee of Virginia Commonwealth University in accordance with the National Institute of Health Guide for the Care and Use of Laboratory
Reagents and supplies
The d3-tetrahydrocannabinol (d3-THC) was purchased from Cerilliant (Texas, USA). Naphthalen-1-yl-(1-pentylindol-3-yl)methanone (JWH-018), Naphthalen-1-yl-(1-butylindol-3-yl)methanone (JWH-073) and 4-chloronaphthalen-1-yl-(1-pentylindolin-3-yl)methanone (JWH-398), Fig. 1, were obtained from John W. Huffman (Clemson University). The methanol, acetonitrile, water and ammonium formate were purchased from Fisher Scientific (New Jersey, USA) and were HPLC grade or better. A 0.1 mg/L JWH-018 and
HPLC/MS/MS validation
The method validation was performed as follows. Validation runs containing calibration standards, blank samples, blank sample with internal standard added and replicates of LLOQ, LQC, MQC and HQC samples were prepared and run on three separate days.
Results and discussion
The presented HPLC/MS/MS method for analysis of blood and brain specimens applied an administrative LOQ for JWH-018 and JWH-073 of 1.0 ng/mL for blood and 4.0 ng/g for brain tissue. The methods of Teske et al. [17] and Sebastian et al. [18] applied to serum analysis were more sensitive with LOQs of 0.5 ng/mL and 0.1 ng/mL, respectively. However, blood and brain are more complex matrices than serum from which to extract and purify lipid soluble compounds such as JWH-018 and JWH-073. Therefore, it
Conflict of interest statement
The authors declare that there are no conflicts of interest.
Acknowledgments
This project was supported by the National Institute on Drug Abuse (NIDA) Center for Drug Abuse Grants R01DA02396, R01DA03672, and P50DA005274.
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Methodology for controlled administration of smoked synthetic cannabinoids JWH-018 and JWH-073
2018, NeuropharmacologyCitation Excerpt :The plasma was certified in-house as drug free by scanning of ammonium hydroxide and methylene chloride liquid-liquid basic extract for drugs by GC/MS operated in scan mode (40–500 m/z) and UPLC/MS/MS for JWH-018, JWH-073 and metabolites prior to use. The extractions were performed using a modification of the procedure of Foltz et al. (1983) as previously described for analysis of JWH-018 and JWH-073 (Poklis et al., 2012). 10 μL of ISTD was added to 1 mL aliquots of calibrators, controls and samples.
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2018, Pharmacology and TherapeuticsApplication of a validated LC-MS/MS method for JWH-073 and its metabolites in blood and urine in real forensic cases
2015, Forensic Science InternationalCitation Excerpt :The reported intoxication cases, the severe adverse effects observed at emergency departments, the increasing popularity of the use of synthetic cannabinoids and the lack of analysis techniques for these chemicals have created an urgent need for the development of new analytical methods. Initially, the abused parent drug was identified in herbal materials [7,14–22] and in biological matrices, such as hair [23–26], oral fluid [27–30], plasma, serum, and whole blood [31–36]. In vitro studies for the identification of phase I and phase II metabolites have been performed [37,38].
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This data was presented, in part, at the Annual Meeting, Society of Forensic Toxicologist, Richmond, VA October 21, 2010.
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Present address: Laboratory Toxicology, Emergency Center Tunis, Tunisia.