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[摘要]
目的 建立HPLC-DVD波长切换联合梯度洗脱法同时测定跌打止痛散(DZS)中6种指标成分羟基红花黄色素A、薯蓣皂苷、原薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷和纤细薯蓣皂苷的量。方法 采用HPLC-DVD法,Diamonsil C18(250 mm×4.6 mm,5 μm)色谱柱;乙腈-0.4%甲酸水溶液为流动相,体积流量0.9 mL/min,梯度洗脱;羟基红花黄色素A的检测波长为403 nm,薯蓣皂苷、原薯蓣皂苷、甲基原薯蓣皂苷、伪原薯蓣皂苷和纤细薯蓣皂苷的检测波长为203 nm;进样量为20 μL。结果 6种指标成分羟基红花黄色素A在3.46~69.20 μg/mL(r=0.999 4)、薯蓣皂苷在9.52~190.40 μg/mL(r=0.999 7)、原薯蓣皂苷在8.74~174.80 μg/mL(r=0.999 6)、甲基原薯蓣皂苷在4.45~89.00 μg/mL(r=0.999 9)、伪原薯蓣皂苷在2.64~52.80 μg/mL(r=0.999 5)、纤细薯蓣皂苷在3.28~65.60 μg/mL(r=0.999 8)质量浓度与峰面积具有较好的线性关系;精密度、重复性良好,RSD均小于2.0%;供试品溶液在室温条件下12 h内稳定;平均加样回收率和相应的RSD分别为98.57%、1.59%;97.64%、1.28%;99.43%、1.07%;97.98%、1.64%;98.57%、1.16%;97.17%、1.37%。结论 建立的HPLC波长切换联合梯度洗脱法同时测定DZS中的6种成分,方法操作简便、快速、准确,可作为DZS全面可靠的质量控制方法。
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[Abstract]
Objective To develop an HPLC-DVD wavelength switching combined with gradient elution method for the determination of the contents of hydroxysafflor yellow A, protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin in Dieda Zhitong San (DZS) simultaneously. Methods The chromatographic separation was achieved on a Diamonsil C18 (250 mm × 4.6 mm, 5 μm) with acetonitrile (A)-0.4% formic acid solution (B) as mobile phase at the flow rate of 0.9 mL/min for gradient elution; Hydroxysafflor yellow A was detected at 403 nm; Protodioscin, dioscin, methylprotodioscin, pseudoprotodioscin, and gracillin were detected at 203 nm; Sample quantity was 20 μL. Results The six active components were well separated and showed good linearity, such as hydroxysafflor yellow A 3.46—69.20 μg/mL (r = 0.999 4), protodioscin 9.52—190.40 μg/mL (r = 0.999 7), dioscin 8.74—174.80 μg/mL (r = 0.999 6), methylprotodioscin 4.45—89.00 μg/mL (r = 0.999 9), pseudoprotodioscin 2.64—52.80 μg/mL (r = 0.999 5), and gracillin 3.28—65.60 μg/mL (r = 0.999 8). The precision and repeatability were good, and RSD values were less than 2.0%. The stability was good in 12 h. The average recoveries and the corresponding RSD values were 98.57% (1.59%), 97.64% (1.28%), 99.43% (1.07%), 97.98% (1.64%), 98.57% (1.16%), and 97.17% (1.37%), respectively. Conclusion An HPLC wavelength switching combined with gradient elution method has been successfully established for simultaneous determination of six components in DZS. The method is simple, quick, accurate, and helpful for the quality control of DZS.
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