Relative response factors and multiple regression models in liquid chromatography to quantify low-dosed components...

A classical quantitative analysis in liquid chromatography is performed using either a one-point calibration or a calibration line, prepared using a reference standard of the compound(s) of interest. However, in some cases, adequate reference standards may be very expensive, rare to obtain, or have limited shelf-life properties. Also, in herbal matrices, multiple compounds could be necessary to be quantified, needing a whole series of different (related) reference standards. In these cases, the use of relative response (sometimes called relative correction factors) factors (RRFs) towards reference standards, different of the compound to be quantified, gained attraction. This study performed a comparison of the use of RRFs and linear relative response factor models (LRRFM) for the quantification of targeted low-dosed compounds using an alternative standard, since it is known that classical RRFs often fail in lower concentration ranges. For this purpose, the determination of the total Δ9-tetrahydrocannabinol (Δ9-THC + Δ9-THC-A) content in dried cannabis flowers, using UHPLC-DAD, was used as a case study. A chromatographic method was implemented and validated, and the use of classical calibration lines, classical RRF, and the LRRFM was applied and compared, with special focus on the concentration around 0.2% (w/w) total Δ9-THC, the legal limit (in most European countries) in these products. Results showed that the newly presented and validated LRRFM approach outperformed the classical RRFs, especially in the low-concentration ranges and that concentrations obtained with the LRRFM were in accordance with the interpolation results obtained with a calibration line.