摘要
目的建立HPLC-ELSD法测定芪芍方有效部位中黄芪甲苷含量和可见分光光度法测定黄芪总皂苷含量的方法,有效控制芪芍方有效部位质量。方法黄芪甲苷含量测定,采用HPLC-ELSD法,Hypersll-C18色谱柱(250mm×4.6mm,5μm),流动相为乙腈-水(31:69),流速1.0mL/min,柱温30℃,漂移管温度101℃,空气流速2.7L/min。黄芪总皂苷含量测定,采用可见分光光度法,MCI柱层析技术分离黄芪总皂苷类,再经反相C18固相萃取小柱纯化,5%香草醛冰醋酸溶液和高氯酸为显色剂,于543nm处测定。结果HPLC法及可见分光光度法中,黄芪甲苷分别在1.30～7.80μg、0.0108～0.065g/L的范围内呈现良好的线性关系,平均加样回收率分别为95.95%、96.58%,RSD分别为2.46%、2.28%。结论本试验建立的方法简便、准确、重复性好,可用于芪芍方有效部位中黄芪皂苷类的质量控制。
Objective To establish a method of HPLC-ELSD for quantitative determination of astragaloside IV and a method of visible spectrophotometry for total astragalosides in the ffective fraction of Qisho Formula,and to control effectively the quality of the effective fraction of Qisho Formula.Methods For the quantity determination of astragaloside IV by HPLC-ELSD,the procedure of HPLC-ELSD was performed on the chromatographic column of Hypersll-C18（250 mm×4.6 mm,5 μm）,and the mobile phase in the gradient elution was acetonitrile-water （31：69）.The flow velocity was 1.0 mL/min.The column temperature was 30 ℃ and drifttube temperature was 101 ℃.The air flow rate was 2.7 L/min.For the quantity determination of total astragalosides by visible spectrophotometry,the column chromatography of MCI was used for separating,and then reversed-phase C18 solid phase extraction （SPE） column,for purification.Vanillin acetic acid solution （5%） and perchloric acid were taken as developing agent for determination at 543 nm.Results Astragaloside IV showed a good linear relationship,respectively,at the range from 1.30 μg to 7.80 μg and from 0.010 8 g/L to 0.065 g/L in HPLC-ELSD procedure and visible spectrophotometry.The average recovery of astragaloside IV was 95.95%（RSD=2.46%） and 96.58%（RSD=2.28%） respectively.Conclusion The methods are easy and accurate with higher repeatability,which can be used in the quality control of astragalosides in the effective fraction of Qisho Formula.
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2010年第4期270-273,共4页
Journal of Beijing University of Traditional Chinese Medicine
