1.School of Pharmacy, Guangdong Pharmaceutical University, Guangzhou 510006, China; 2.State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou 510070, China
Abstract:OBJECTIVE To establish the HPLC fingerprint analysis method for agarwood with compare the natural agarwood and artificial agarwood combined with the contents of ethanol-soluble extraction (E%) and agarotetrol (A%). METHODS The E% and A% were determined according to the ChP(2015). The chromatographic fingerprints of agarwood were established on a Platisil ODS C18 column (4.6 mm×250 mm, 5 μm) with mobile phase consisting of acetonitrile-0.1% formic acid. Gradient elution was performed at a flow rate of 0.7 mL·min-1, the detection wave length was set at 252 nm, and the column temperature was maintained at 31 ℃. The similarity evaluation system of traditional Chinese medicine chromatographic fingerprint (2004A) was used to establish the common pattern and calculate the similarity of chromatograms. Principal component analysis (PCA) was used with multivariate statistical analysis software for relative peak area of common chromatographic peak. RESULTS The chromatographic fingerprint similarity of 13 batches of natural agarwood was 0.021-0.856, the E% was 10.1%-31.0%, and the A% was 0.04%-2.83%, respectively. The chromatographic fingerprint similarity of 11 batches of artificial agarwood ranged from 0.079 to 0.453, and the E% and A% were 10.3%-31.3% and 0.14%-1.02%, respectively. Thirteen batches of natural agarwood and 11 batches of artificial agarwwod were divided into two groups respectively, and the reference crude drug was gathered in the natural agarwood group. The similarity between natural and artificial agarwood chromatographic fingerprint was significantly different (P<0.05). Meanwhile, the similarity of artificial agarwood chromatographic fingerprint was positively correlated with the E% (P<0.05). CONCLUSION The chromatographic fingerprint analysis combined with multivariate statistical analysis can easily and quickly distinguish natural and artificial agarwood, which provides a reference for the comprehensive evaluation of the quality of agarwood.
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