六神丸HPLC-UV/ELSD指纹图谱研究

目的?建立六神丸的HPLC-UV/ELSD指纹图谱分析方法，对六神丸中主要特征峰进行归属并进行指认。方法?采用HPLC法，色谱柱为Kromasil C₁₈（250?mm×4.6?mm，5?μm）；乙腈-0.05%三氟乙酸水溶液作为流动相，进行梯度洗脱；紫外检测器（UV）参数:检测波长为296?nm；蒸发光散射检测器（ELSD）参数:漂移管温度为70?℃，喷雾器为冷却模式，氮气压力为25?psi；进样量20?μL；体积流量为1?mL/min。通过《中药色谱指纹图谱相似度评价系统（2012版）》，建立14批六神丸指纹图谱的共有模式，计算其相似度，并对共有峰进行药材的归属以及成分进行指认。结果?分别建立了六神丸HPLC-UV及HPLC-ELSD的指纹图谱，及其共有模式，指认出HPLC-UV指纹图谱共有峰14个，14批六神丸制剂相似度为0.940～0.988，指认出HPLC-ELSD指纹图谱共有峰13个，14批六神丸制剂相似度为0.968～0.998。确认了和蟾蜍他灵、沙蟾毒精、远华蟾毒精、蟾毒它灵、华蟾毒它灵、蟾毒灵、胆酸、鹅去氧胆酸、去氧胆酸。结论?首次建立了六神丸HPLC-UV/ELSD指纹图谱分析方法，该法操作准确、简单，重复性好、精密度高，可用于分析六神丸中化学成分信息，为六神丸质量控制提供了可靠的科学依据。 

HPLC-UV/ELSD Fingerprint of Liushen Pills

OBJECTIVE?HPLC-UV/ELSD fingerprint was developed to assign and identify the main characteristic peaks in Liushen Pills. METHODS?High performance liquid chromatography: Kromasil C₁₈ column (250?mm × 4.6?mm， 5?μm) was used with a gradient mobile phase system of acetonitrile-water containing 0.05% trifluoroacetic acid. Ultraviolet detector: the detection wavelength was 296?nm; Evaporative light scattering detector: The temperature of drift tube was maintained at 70?℃， sprayer parameter was cooling mode and nitrogen pressure was 25?psi. The injection volume was 20?μL; The flow rate was 1?mL/min. The similarities between the HPLC-UV/ELSD fingerprints of 14 batches of Liushen Pills were calculated by similarity evaluation software， and the common peaks were assigned and identified simultaneously. RESULTS?The chromatography fingerprint of Liushen Pills from 14 different origins was established. In the chromatography fingerprint with HPLC-UV of Liushen Pills， 14 common peaks were damarcated and the similarities of Liushen Pills were between 0.940-0.988， and the chromatography fingerprint with HPLC-ELSD of Liushen Pills， 13 common peaks were damarcated and the similarities of Liushen Pills were between 0.968-0.998. Gamabufotalin， arenobufagin， telocinobufagin， bufotalin， cinobufotalin， bufalin， cholic acid， chenodeoxycholic acid， deoxycholic acid were identified. CONCLUSION?It is the first time to establish the HPLC-UV/ELSD fingerprint of Liushen Pills. The method is simple， accurate， and reproducible， which can be used to characterize the chemical composition information of Liushen Pills and provide a scientific evidence for the quality control of Liushen Pills.