HPLC fingerprint and determination of two components in Zhisou Powder
OBJECTIVE To establish a high performance liquid chromatography(HPLC)fingerprint of Zhisou Powder and validate a method for determining the content of two components.METHODS Ten batches of Zhisou Powder samples were pre-pared.SunFireTM C18column(250 mm × 4.6 mm,5 μm)was employed by gradient elution with a mobile phase comprising of acetonitrile and water(containing 0.1％phosphoric acid)at a flow rate of 1.0 mL·min-1.Injection volume was set at 10 μL per injection with variable wavelength detection.The detection wavelengths were 320 nm(0-4 min),280 nm(4-45 min)and 254 nm(45-90 min).Column temperature was 30 ℃.The HPLC fingerprint of 10 batches of Zhisou Powder samples was estab-lished.Similarity analysis was evaluated by the evaluation software for chromatographic fingerprints of traditional Chinese medi-cine(version 2012A).Hesperidin and ammonium glycyrrhizinate were utilized two index components for content analysis.RESULTS A total of 17 common peaks were identified in 10 batches of Zhisou Powder fingerprints with two reference compo-nents of hesperidin and ammonium glycyrrhizinate.HPLC fingerprints of 10 batches of Zhisou Powder were established.Similar-ity had a range of 0.978 to 1.000 and 17 common peaks were confirmed.Hesperidin and ammonium glycyrrhizinate showed a decent linear relationship at 105.9-10 590 μg·mL-1 and 22.96-229.6 μg·mL-1(R2≥0.999 6).The precision,repeatability,stability and sample recovery tests were all promising.CONCLUSION The established Zhisou Powder fingerprint and content determination method are stable,reliable and reproducible.It provides scientific rationales for quality control of Zhisou Powder.
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