Detection and disposition of JWH-018 and JWH-073 in mice after exposure to “Magic Gold” smoke

https://doi.org/10.1016/j.forsciint.2012.02.003Get rights and content

Abstract

The disposition in mice of the cannabimimetics JWH-018 and JWH-073 in blood and brain following inhalation of the smoke from the herbal incense product (HIP) “Magic Gold” containing 3.6% JWH-018, 5.7% JWH-073 and less than 0.1% JWH-398 (w/w) is presented. Specimens were analyzed by HPLC/MS/MS. The validation of the method is also presented. Five C57BL6 mice were sacrificed 20 min after exposure to the smoke of 200 mg of “Magic Gold” and a second set of five exposed mice were sacrificed after 20 h. Twenty minutes after exposure to “Magic Gold” smoke, blood concentrations of JWH-018 ranged from 42 to 160 ng/mL (mean: 88 ng/mL ± 42) and those of JWH-073 ranged from 67 to 244 ng/mL (mean: 134 ng/mL ± 62). Brain concentrations 20 min after exposure to “Magic Gold” smoke for JWH-018 ranged from 225 to 453 ng/g (mean: 317 ng/g ± 81) and those of JWH-073 ranged from 412 to 873 ng/g (mean: 584 ng/g ± 163). Twenty hours after exposure to “Magic Gold” smoke, JWH-018 was detected and quantified in only two of the five blood samples. Blood concentrations of JWH-018 were 3.4 ng/mL and 9.4 ng/mL. JWH-073 was detected in only one blood specimen 20 h after exposure at 4.3 ng/mL. Brain concentrations 20 h post exposure for JWH-018 ranged from 7 to 32 ng/g (mean: 19 ng/g ± 9). JWH-073 was not detected in 20 h post exposure brain specimens. JWH-398 was not detected in any of the blood or brain samples. The disposition data presented with the limited data available from human experience provide reasonable expectations for forensic toxicologists in JWH-018 or JWH-073 cases. As with THC after smoking marijuana, blood and brain concentrations of JWH-018 and JWH-073 after HIP smoking can be expected to rise initially to readily detected values, and then drop dramatically over the next few hours to several ng/mL or ng/g, and finally to be at extremely low or undetectable concentrations by 24 h apparently due to extensive biotransformation, and redistribution to body fat.

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.

References (28)

  • European Monitoring Centre for Drugs and Drug Addiction, Action on New Drugs Briefing Paper: Understanding the Spice...
  • V. Auwärter et al.

    Spice and other herbal blends: harmless incense or cannabinoid designer drugs?

    J. Mass Spectrom.

    (2009)
  • N. Uchiyama et al.

    Identification of a cannabinoid analog as a new type of designer drug in a herbal product

    Chem. Pharm. Bull.

    (2009)
  • N. Uchiyama et al.

    Identification of a cannabimimetic indole as a designer drug in a herbal product

    Forensic Toxicol.

    (2009)
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