A Fast, Sensitive, Comprehensive and Accurate Quantitative Assay of 16 Cannabinoids in Different Matrix by LC/MS/MS



In 2018, AOAC has published a method to quantify 10 cannabinoids (CBDs) with the range of 0.500-10.0 µg/mL in cannabis. However, there are more than 100 CBDs isolated from cannabis in addition to above 10 CBDs. Furthermore, CBDs have been legally and widely used for more and more products. Thus, AOAC method may not cover various needs from different CBD products. In order to cover various needs, Dyad Labs has developed UPLC-PDA and UPLC-MS/MS methods. In this poster, the fast, sensitive, accurate and comprehensive quantitative LC/MS/MS method for 16 major CBDs was presented in different matrix. The UPLC-PDA method is presented in P-T-019 poster.


Sample Preparation and Extraction:

Appropriate sample was hydrated with 5 mL of DI water, and then extracted with 35 mL of methanol. The sample is then diluted if needed, and then cleaned up with filter. Sample is diluted with equal volume of water to match with LC condition.

LC-MS/MS Conditions

LC system: Nexera UPLC system including SIL-30AC auto-sampler, controller, column heater and binary pump (Shimadzu)

MS detector:  Triple Quadrupole 5500 MS (AB Sciex)

Results and Discussions

Chromatographic Separation

Some CBDs are isomers and have same MRM transitions, thus chromatographic separation is very critical in this method. We have investigated on different columns, mobile phases and buffers to achieve enough separation, sharp and symmetric peak shape. A special C18 column provided sufficient separation.

Specificity, Sensitivity and Linearity

The specificity results indicated that there is no interference between analyte, IS and in blank matrix.  The curve range of 20.0-2000 ng/mL was successfully validated. The regression is quadratic with 1/x as the weighing factor. The correlation coefficient R2 is > 0.995. The representative chromatograms for LLOQ and ULOQ samples were in Figure 3.

Accuracy and Precision

The accuracy and precision were investigated in CBD oil sample and with post-spiking pesticides in blank botanical and non-botanical matrix at lower, medium and high regions of the established range of the calibration curve.


This method is  a fast, sensitive, comprehensive and accurate method, and the first validated method to simultaneously quantify 16 major CBDs in in CBD oil, botanical and protein matrix using UPLC-MS/MS.