摘要
目的:建立丹参配方颗粒中丹参素、原儿茶醛、咖啡酸、迷迭香酸、紫草酸及丹酚酸B 6个成分的双标多测法,验证该方法在丹参配方颗粒质量评价中应用的准确性和可行性。方法:采用HPLC法,以乙腈为流动相A,以0.05%的磷酸溶液作为流动相B,梯度洗脱(0~15 min,10%A→20%A;15~40 min,20%A→25%A;40~50 min,25%A→30%A;50~50.1 min,30%A→10%A;50.1~60 min,10%A),流速1 mL·min^(-1),柱温40℃,检测波长286 nm,进样量10μL。采用4根型号不同的C18色谱柱,测定丹参配方颗粒6个成分的实际保留时间,计算各成分在各色谱柱保留时间的平均值,得到各成分的标准保留时间。选取原儿茶醛(峰2)和迷迭香酸(峰4)作为双标化合物,使用双标多测法准确定位各成分色谱峰,预测成分在未知色谱柱的保留时间并进行方法学验证。以迷迭香酸为参照物,计算其余各成分的相对校正因子,对各成分进行定量并与外标法所测结果进行比较。结果:收集的6批样品中丹参素、原儿茶醛、咖啡酸、迷迭香酸、紫草酸、丹酚酸的含量范围分别为3.82~6.54 mg·g^(-1)、0.44~1.14 mg·g^(-1)、42.75~179.49 mg·g^(-1)、2.20~2.63 mg·g^(-1)、3.11~3.49 mg·g^(-1)、42.41~53.74 mg·g^(-1)。双标线性校正法能够准确地预测待测组分在未知色谱柱上的保留时间,且预测结果与实际值的偏差小于相对保留时间法。使用相对校正因子法和外标法的含量结果无显著性差异。结论:双标多测法同时测定丹参配方颗粒中6个酚酸类成分可行且准确,且该法操作简便,准确度高,经济适用,为丹参配方颗粒中6个酚酸类的含量测定提供了新的思路。
Objective:To establish a two reference substances for determination of multiple component method for six components(salvianic acid A,protocatechuic aldehyde,caffeic acid,rosmarinic acid,lithospermic acid and salvianolic acid B)of Danshen formula granules,and to verify the accuracy and feasibility of this method in hequality evaluation of Danshen formula granules.Methods:HPLC method was used with acetonitrile as mobile phase A,using 0.05%phosphoric acid solution as mobile phase B,and gradient elution(0-15 min,10%A→20%A;15-40 min,20%A→25%A;40-50 min,25%A→30%A;50-50.1 min,30%A→10%A;50.1-60 min,10%A),The flow rate was 1 mL·min^(-1),column temperature was 40℃,detection wavelength was 286 nm,and injection volume was 10μL.Four different types of C18 chromatographic columns were used to measure the actual retention time of the six components of Danshen formula granules,and the average value of the retention time of each component in each chromatographic column was calculated to obtain the standard retention time of each component.Protocatechuic aldehyde(peak 2)and rosmarinic acid(peak 4)were selected as the reference substances for the determination.The chromatographic peaks of each component were accurately located by linear calibration using two reference substances,and the position of each component in unknown chromatographic col⁃umn could be predicted in the same method.The methodology was validated.Taking rosmarinic acid as reference,the relative correction factors of the remaining components were calculated,and each component was quantified and compared with the results measured by the external standard method.Results:The contents of salvianic acid A,protocatechuic aldehyde,caffeic acid,rosmarinic acid,shikonic acid and salvianolic acid B in the collected 6 batches of samples were 3.82-6.54 mg·g^(-1),0.44-1.14 mg·g^(-1),42.75-179.49 mg·g^(-1),2.20-2.63 mg·g^(-1),3.11-3.49 mg·g^(-1),42.41-53.74 mg·g^(-1),respectively.The double-standard linear correction method can accurately predict the retention time of the component to be tested on the unknown chromatographic column,and the deviation between the predicted result and the actual value was smaller than that of the relative retention time method.There was no significant difference between the content results using the relative correction factor method and the external standard method.Conclusion:The simultaneous determination of 6 phenolic acids in Danshen formula granules was feasible and accurate by two reference substances for determination of multiple components method,and the method was simple,accurate,economical,providing a new idea for the content determination of 6 phenolic acids in Danshen formula granules.
作者
纪国力
周伟
巴然然
林翠华
刘玉玲
JI Guo-li;ZHOU Wei;BA Ran-ran;LIN Cui-hua;LIU Yu-ling(Tai’an Institute for Food and Drug Control/Tai’an Fiber Inspection Institute,Tai’an 271000,China;School of Pharmacy,Shandong First Medical University(Shandong Academy of Medical Sciences),Tai’an 271000,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2023年第8期1326-1333,共8页
Chinese Journal of Pharmaceutical Analysis
基金
2021年山东省药品质量风险监测资助项目
关键词
丹参配方颗粒
双标多测法
相对校正因子
高效液相色谱法
丹参素
原儿茶醛
咖啡酸
迷迭香酸
紫草酸
丹酚酸B
Danshen formula granules
two reference substances for determination of multiple component method
relative correction factor
HPLC
salvianic acid A
protocatechuic aldehyde
caffeic acid
rosmarinic acid
litho⁃spermic acid
salvianolic acid B
