Confirming the presence of cannabinoids in Cannabis sativa by HPTLC-MS

Introduction

In recent years a lot of research has been done on the medicinal use of Cannabis sativa. Cannabiol (CBN), cannabidiol (CBD) and Δ9-tetrahydrocannabinol (Δ9-THC) are the most studied active ingredients for medicinal use treating several health disorders, e.g. migraine, epilepsy, appetite loss. In addition to its medicinal use cannabis has been legalized by several countries as marihuana.

Fig. 1 Plate under white light (remission); left: plate derivatized with Fast Blue salt B reagent, right: not derivatized part of the plate after elution of zones of interest

Scope

This method is suitable for confirming the presence of the cannabinoids CBN, CBD and THC in Cannabis sativa. The TLC-MS Interface 2 (CAMAG) is used to directly elute target zones from the HPTLC plate into the Waters ACQUITY QDa^® for mass detection. This HPTLC-MS method offers a rapid way for substance confirmation, which can be used for screening and for quality control. Several samples can be separated in parallel and only the zones of interest are analyzed, generating less data.

Required or recommended devices

Automatic TLC Sampler 4 or Linomat 5, Automatic Developing Chamber ADC 2, Immersion Device 3, TLC Visualizer, visionCATS software, TLC-MS Interface 2, Waters ACQUITY QDa Detector (Performance), Empower^® or MassLynx^® software, optional: smartCut, UV Cabinet, TLC Scanner.

Derivatization reagent

Reagent name: Fast Blue salt B Reagent preparation: Weigh 1 g of fast blue salt B (o-dianisidine bis(diazotized) zinc double salt) into a glass bottle and dissolve it in 200 mL of water.

Sample

500 mg of each loosely ground sample is mixed with 10 mL of methanol, sonicated for 10 minutes, and then centrifuged for 5 min (2750 RCF). The supernatant is collected and used as test solution. Cannabis oil and cannabis tincture are diluted 1:10 with methanol.

Standards

Standards were provided by Lipomed AG (Arlesheim, Switzerland), CBN and CBD with a concentration of 1 mg/mL in methanol and Δ9-THC with a concentration of 1 mg/mL in ethanol. Standards are diluted 1:10 with methanol.

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