Rapid Method for Simultaneous Determination of γ-Oryzanol Compounds in Rice (Oryza sativa) Grains using UV-Vis Spectroscopy and Chemometrics

Rice (Oryza sativa) contains γ-oryzanol, which consists of 4 main compounds: Cycloartenol ferulate, 2,4-methylenecycloartanyl ferulate, campesterol ferulate, and β-sitosterol ferulate, that contribute to thehealth benefits of rice. This research aimed to develop a rapid method to determine the 4 major γ-oryzanolcompounds in 45 varieties covering pigmented (black and red) and non-pigmented (white) rice grain. The method was developed by integrating UV-Vis spectroscopy with chemometrics, specifically principal component analysis (PCA) and partial least square (PLS) regression. Sampling was performed across the Indonesian archipelago collecting 180 samples, which comprises 60 samples for each type of rice in form of rice husk, bran, whole grain, and polished rice. The results of PCA on the spectroscopic data successfully identified distinction for the 3 types of rice attributable by different type and levels of chemical compoundin the grain. White rice exhibited a characteristic absorption at 325 nm while in pigmented (red and black) rice showed a maximum absorbance at 280 nm, indicating the presence of different composition of chemical compounds. A reliable model to predict the 4 major γ-oryzanol compounds was established with R2 calibration, R2 cross-validation, and R2 prediction higher than 0.9 was obtained using the spectroscopic data in the range from 200 to 400 nm. However, the PLS modeling was unsuccessful for red and black ricesamples most likely due to the interfering high absorption of red colored compounds. Compared to theexisting techniques for analyzing individual compounds of the γ-oryzanol by high-performance liquid chromatography, the newly developed approach using UV-Vis spectroscopy combined with chemometrics is more practical, faster, and cost-efficient and mainly, solvent and residues free.