Study on HPLC Fingerprint,Content Determination,and Chemical Pattern Recognition of Qinggan Runfei Zhike Distillate
Objective To establish the HPLC fingerprint of Qinggan Runfei Zhike Distillate and determine the contents of morphine,codeine,papaverine and narcodine,and carry out chemical pattern recognition in order to evaluate the quality of the preparation comprehensively.Methods HPLC was carried out by using methanol-5 μmoL·L-1 sodium heptanesulfonate solution(adjust pH to 2.6 with phosphoric acid)as the mobile phase for gradient elution.The flow rate was 1.0 mL·min-1,the column temperature was 20℃,the detection wavelength was at 220 nm,and the injection amount was 10 μL.Similarity evaluation combined with hierarchical clustering analysis(CA),principal components analysis(PCA)and discriminant analysis of partial least squares(PLS-DA)were used to evaluate the HPLC chromatographic fingerprint of 10 batches of Qinggan Runfei Zhike Distillate.Results There were 16 common peaks in the fingerprint of 10 batches of Qinggan Runfei Zhike Distillate,and 5 common peaks were identified,which were morphine,codeine,thebaine,papaverine and nacodine.The linear range of morphine,codeine,papaverine and nacodine in the content determination results are 0.70-8.70,0.68-8.50,0.60-7.55 μg,0.38-4.80 μg(r≥0.999 7).The RSDs of precision,stability and repeatability tests are less than 3.0%.The average sample recovery rate ranges are 97.93%-99.43%(RSD＜3.1%).The contents in Qinggan Runfei Zhike Distillate are morphine 43.58-76.47 μg·mL-1,codeine 16.64-78.92 μg·mL-1,papaverine 29.93-41.22 μg·mL-1,nacodine 14.44-33.22 μg·mL-1.The results of CA,PCA and PLS-DA showed that ten batches of Qinggan Runfei Zhike Distillate could be clustered into two categories,S3,S4,S8,S10 were gathered into one category,and the rest were clustered into another category.Nine differential components were screened out,including peak 2,peak 4,peak 13,peak 11(morphine),peak 1,peak 10,peak 8,peak 12,and peak 16(nacodine).Conclusion The HPLC fingerprint and content determination method established in this study are simple and accurate.The above method combined with chemical pattern recognition can be used for the quality control of Qinggan Runfei Zhike Distillate.
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