Development and Validation of the LC–MS/MS Method for Determination of 130 Natural and Synthetic Cannabinoids in Cannabis Oil

Abstract

Dietary supplements are widely available products used by millions of people around the world. Unfortunately, the procedure of adding pharmaceutical and psychoactive substances has recently been observed, in order to increase the effectiveness of supplements in the form of hemp oils. For this reason, it is extremely important to develop analytical methods for the detection of substances prohibited in dietary supplements and food products. In the present study, using the LC–MS/MS technique, an innovative method for the detection and quantification of 117 synthetic cannabinoids and 13 natural cannabinoids in dietary supplements and food products in the form of oils during one 13-min chromatographic run was developed. Each method was fully validated by characterization of the following parameters: The limit of detection was set to 0.1 ng/mL (100 µg/g, 0.01%). The limit of quantification ranged from 0.05 ng/mL to 50 ng/mL. The criteria assumed for systematic error caused by methodological bias (±20%) resulting from the recovery of analytes after the extraction process, as well as the coefficient of variation (CV) (≤20%), were met for all 130 tested compounds. The positive results of the validation confirmed that the developed methods met the requirements related to the adequacy of their application in a given scope. Additionally, methods developed using the LC–MS/MS technique were verified via proficiency tests. The developed analytical procedure was successfully used in the analysis of hemp oils and capsules containing them in the studied dietary supplements.

Progress in the analysis of phytocannabinoids by HPLC and UPLC (or UHPLC) during 2020–2023

“…[1][2][3][4] There are several studies on the HPLC-based analysis of cannabis oil or CBD (5)-rich oil available in the literature for the period covered in this article. 44,97,[102][103][104][105][106][107][108][109][110][111][112] However, the analysis of cannabis oil using UHPL or UPLC has been limited to a few studies reported in the literature (Table 2).…”

“…Analakkattillam et al 108 reported an isocratic HPLC run (20 min) for the detection of CBD (5) and Δ 9 -THC (14) in cannabis oil-infused products, while a gradient elution was conducted with water and MeOH, both containing 0.1% HCOOH and 2 mM ammonium formate, to identify CBC (1), CBD (5), CBDA (6), CBDVA (18), CBG (8), CBGA (9), cannabigerovarinic acid (CGBVA), CBN (10), cannabivarin (CBV), Δ 9 -THC ( 14), Δ 9 -THCA (15), THCV (16), and THCVA (17) in cannabis oil and oil-containing capsules using an ESI-MS detector operated in positive ion mode. 109 The use of the isocratic mobile phase in HPLC-UV (or PDA) analysis of CBD-oil has been quite popular as the analysis generally targets the detection of CBD ( 5) and a few other cannabinoids. 44,97,111 Li et al 111 have recently reported the analysis of CBD ( 5) full spectrum oil revealing the presence of CBD ( 5), CBDA (6), CBDV (18) (5), CBDA (6), CBN (10), Δ 9 -THC ( 14), and Δ 9 -THCA (15) in the samples using a simple UV detector.…”

“…Since the publication of the review article 2 on the application of HPLC and UPLC (or UHPLC) methods in the analysis of phytocannabinoids covering the period of 2010–2019, several publications have appeared in the literature on this topic 12–185 . All these publications highlight the importance of HPLC and UPLC (or UHPLC) techniques as well as showcase remarkable advancements in sensitivity and versatility associated with these techniques in relation to phytocannabinoid analysis 12 .…”

“…A large volume of HPLC and UPLC (or UHPLC) methods that have been reported for the analysis of cannabinoids in C. sativa L. plant samples 13–94 and cannabis consumer products, for example, hashish, marijuana, and cannabis oils, 95–137 since 2020 are presented in Tables 1 and 2 and discussed in the following subsections.…”

“…These consumer products include smoking products, for example, hashish and marijuana, cannabidiol oil, cannabis oil, cannabis olive oil, infusions and liquids, tinctures, cannabis food products, and cannabis‐containing cosmeceuticals (Table 2). 95–137 In the HPLC and UPLC (or UHPLC) methods, the use of PDA or MS detection technologies has become routine for the analysis of these consumer products 2 . The use of HPLC as opposed to UPLC (or UHPLC) has remained the method of choice during 2020–2023, but the use of UPLC (or UHPLC) has increased considerably over the last few years (Table 2).…”