甲肿消制剂及其含药血清中哈巴俄苷含量测定.docVIP

甲肿消制剂及其含药血清中哈巴俄苷含量测定 　 作者：张秋燕，崔翰明，刘喜明，朱晓芸，白鸽 【摘要】 目的建立甲肿消制剂及灌胃给予甲肿消后大鼠血清中的哈巴俄苷含量测定方法。方法采用RP-HPLC法测定哈巴俄苷的含量，色谱柱：Kromasil C18色谱柱（250 mm×4.6 mm，5 μm）和Easyguard保护柱，以乙腈和1％醋酸溶液为流动相，采用梯度洗脱；流速：1 ml· min-1；检测波长为278 nm；进样量：5 μl。结果哈巴俄苷在1.46～146 μg·ml-1的浓度范围内线性良好；低、中、高浓度哈巴俄苷的精密度（RSD）(n=5)分别为1.05%，0.59%，0.80%；日间差（RSD）（n=5）为1.00%；制剂和大鼠血清中哈巴俄苷的加样回收率（n=5）分别为（98.02±1.01）%和（101.03±1.64）%；测定其重现性（RSD）（n=6）分别为0.73%和0.96%；每克制剂中含哈巴俄苷的量为230.5 μg，每毫升含药血清中哈巴俄苷的量为1.88 μg。结论该方法简便、快速、准确、重现性良好，可用于制剂和血清中哈巴俄苷的含量测定。 【关键词】 哈巴俄苷； 甲肿消制剂； 固相萃取； 反相高效液相色谱 ChinaAbstract：ObjectiveTo develop a determination method of Harpagoside in Jia Zhong Xiao preparation and blood serum of rat. Methods The RP-HPLC method was applied as with: Kromasil C18 column（250 mm×4.6 mm，5 μm） and Easyguard column. The mobile phase was acetonitrile and 1% acetic acid with linearity gradient elution, the flow rate at 1 ml·min-1, and detection wavelength was 278nm. ResultsAn excellent linearity was obtained over the range of 1.46～146 μg·ml-1. The RSDs of accuracy for 7.3, 29.2, 146μg·ml-1 were 1.05%,0.59%,0.80%, respectively (n=5). The RSDs of intra-day validation were 1.00%. The mean recovery rates of Harpagoside in preparation and serum were (98.02±1.01)%, (101.03±1.64)%,respectively. The RSDs of the determination method for preparation and serum repeatability were 0.73% and 0.96%, respectively. The content of Harpagoside in preparation and serum was 230.5μg·g-1 and 1.88μg·ml-1, respectively.ConclusionThe method is simple, rapid, reliable and repeatable. It’s quite suitable for the analysis of Harpagoside in Jia Zhong Xiao preparation and of rat serum. 　　Key words：Harpagoside; Jia Zhong Xiao preparation; SPE; RP-HPLC 甲肿消处方由玄参、夏枯草等组成，按适当工艺制备成浓度为3g·ml-1饮片的浸膏剂。动物和临床试验证明本制剂具有较好的化痰散结、治疗甲亢型甲状腺肿大的作用。处方中玄参具有凉血滋阴、泻火解毒的功效，用于热病伤阴、津伤便秘、目赤、咽痛等症［1］，其含量较高的特征性成分为哈巴俄苷，有抗慢性炎症、降压、镇痛、解痉、抗乙型肝炎病毒和免疫促进等作用［2，3］。因此，我们采用反相高效液相色谱法建立了浸膏和大鼠血清中的哈巴俄苷的含量测定方法，为进一步阐明本方治疗甲亢的机理和控制制剂质量提供分析方法。 　　1 仪器与材料 　　1.1 仪器Knaver 液相色谱仪、Knaver UV Detecter（德国Knaver公司）；Kromasil C18