Study on HPLC Fingerprint of Bombyx mori from Different Producing Areas and Content Determination of Multi- component / 中国药房

ABSTRACT

OBJECTIVE：To establish the fingerprint of Bombyx mori and the method for the content determination of multi- components，and to provide reference for comprehensive quality evaluation of B. mori . METHODS ：Using 18 batches of B. mori from different producing areas as samples ，HPLC method was used. The column was Shiseido CAPCELL PAK C 18 AQ S 5 with mobile phase consisted of methanol- 0.05 mol/L potassium dihydrogen phosphate solution （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was 260 nm，and column temperature was set at 30 ℃. The sample size was 10 μL. HPLC fingerprint analysis and similarity evaluation were performed by using TCM Chromatogram Fingerprint Similarity Evaluation System（2012 edition），and the chromatographic peak was identified by comparing with the chromatogram of reference substance. The contents of 4 nucleosides as uracil ，guanine，xanthine，uridine were determined . RESULTS ：A total of 16 common peaks were identified in HPLC fingerprint for 18 batches of B. mori ，and peaks 3，6 ，7 and 8 were identified as uracil ，guanine，xanthine and uridine. The similarity of sample chromatogram with control fingerprint were 0.912-1.000. The linear range of uracil ，guanine， xanthine and uridine were 5.34-534，5.28-528，5.06-506，5.195-519.5 μg/mL（r≥0.999 8）. The limits of detection were 0.032 4， 0.032 0，0.030 7，0.031 2 μg/mL，and the limits of quantitation were 0.106 8，0.105 6，0.101 2，0.103 0 μg/mL. RSDs of precision，reproducibility and stability tests （24 h）were all lower than 1.00％（n＝6）. Average recoveries were 100.15％-102.95％， and RSD s were all lower than 2.00％（n＝9）. The content determination results showed that the content of uracil ，guanine， xanthine and uridine of B. mori from different producing areas were 0.41％-2.46％，0.37％-1.98％，0.72％-2.63％，0.94％-3.67％， respectively. CONCLUSIONS ：Established HPLC fingerprint and content determination method of 4 nucleosides were specific ， accurate and reliable ，which can be used for the quality evaluation and control of B. mori .