Quality Evaluation of raw Oryzae Fructus Germinatus Based on Fingerprint, Pattern Recognition and Multi-components Determination

OBJECTIVE To establish the fingerprint of raw Oryzae Fructus Germinatus by HPLC, analyze them with chemical pattern recognition technology, and determine the contents of 5-hydroxymethylfurfural, protocatechuic acid, vanillic acid, p-coumaric acid and ferulic acid in raw Oryzae Fructus Germinatus, in order to provide a scientific basis for its quality evaluation.

METHODS The HPLC fingerprint of raw Oryzae Fructus Germinatus was performed on Ultimate AQ-C18(250 mm×4.6 mm, 5 μm) column with acetonitrile-0.3% phosphoric acid solution for gradient elution, the fingerprint data was analyzed by the similarity evaluation system of traditional Chinese medicine chromatographic fingerprints combined with cluster analysis and principal component analysis. Furthermore, the contents of five components in raw Oryzae Fructus Germinatus were determined.

RESULTS The HPLC fingerprint of 16 batches of raw Oryzae Fructus Germinatus was established. The similarity were all≥0.891. Eighteen common peaks were marked and 5 of them were identified, including 5-hydroxymethylfurfural(peak 3), protocatechuic acid(peak 4), vanillic acid(peak 10), p-coumaric acid(peak 17) and ferulic acid(peak 18). Cluster analysis and principal component analysis were used to classify the 16 batches of raw Oryzae Fructus Germinatus into four groups. The results of multi-component quantitative analysis methodology showed that the linear relationship(R²≥0.9993) of five components were good, the average recoveries were between 99.12% and 101.37%, and the RSD were in the range of 0.02%−2.40%. The contents of 5-hydroxymethylfurfural, protocatechuic acid, vanillic acid, p-coumaric acid and ferulic acid in 16 batches of raw Oryzae Fructus Germinatus were in the range of 0.36−3.57, 1.42−23.36, 6.60−39.37, 33.02−72.33, 29.72−72.59 μg·g⁻¹, respectively.

CONCLUSION The method of HPLC fingerprint combined with pattern recognition and multi-components simultaneous determination is rapid, simple and reproducible, which can provide a reference for the quality evaluation of raw Oryzae Fructus Germinatus.