HPLC-DAD method development and validation for the quantification of CBD, CBDA, Δ9-THC AND Δ9-THCA in Cannabis-based...

The therapeutic use of Cannabis has significantly expanded in recent years, increasing the demand for robust, accessible, and reliable analytical methods capable of ensuring the quality, safety, and efficacy of the plant-derived products. In this context, the present study aimed to develop and validate a chromatographic method using HPLC-DAD for the identification and quantification of four cannabinoids (CBD, CBDA, Δ⁹-THC, and Δ⁹-THCA) in different matrices - oil and marijuana. Method validation confirmed its selectivity, linearity, precision, and accuracy, in accordance with the criteria established by applicable guidelines. Recovery values ranged from 81.2% to 107.9% for oil samples and from 80.3% to 107.7% for Cannabis plant material. Limits of detection (LOD) ranged from 0.0258 µg mL^-1 to 0.2043 µg mL^-1, while limits of quantification (LOQ) varied from 0.0851 µg mL^-1 to 0.6744 µg mL^-1. Intermediate precision ranged from 2.5% to 7.8% for oil and from 6.0% to 10.9% for Cannabis plant material. Repeatability ranged from 2.7% to 9.4% for oil, and from 5.5% to 9.1% for Cannabis plant material. The developed method was successfully applied to three samples, enabling the identification of one CBD-rich oil and two Δ⁹-THC/Δ⁹-THCA-predominant samples, one oil and one plant material. The results demonstrate that the method is suitable for quality control of Cannabis-based products and can be effectively applied to both oil and plant-based matrices.