摘要
目的:建立乳块消片中5个成分(丹参素、王不留行黄酮苷、橙皮苷、丹酚酸B、丹酚酸A)的含量测定方法,探索双标多测法(双标线性校正法用于色谱峰定性以及采用相对校正因子定量)的可行性。并以5个成分的含量测定为例,比较双标线性校正法和相对保留时间法2种定性方法,以及相对校正因子法和常规外标法2种定量方法的优劣和差异。方法:采用HPLC法,以乙腈为流动相A,0.2%磷酸水溶液为流动相B,梯度洗脱(0~30 min,5%A→20%A;30~40 min,20%A→30%A;40~50 min,30%A),流速为1 mL·min-1,检测波长为282 nm,柱温为30℃。在4根不同的C18色谱柱上测定乳块消片5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,选取丹参素(峰1)和橙皮苷(峰3)2个成分作为双标化合物,使用双标线性校正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。以中间峰橙皮苷为参照物,计算橙皮苷与其余各成分的相对校正因子,对各成分进行定量。结果:不同企业的样品中丹参素、王不留行黄酮苷、橙皮苷、丹酚酸B、丹酚酸A的含量范围分别为3.14~6.25、0.37~1.79、0.42~1.18、0.91~3.50、1.40~6.97 mg·g-1。双标线性校正法能够准确地预测待测成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,对色谱峰的定位比相对保留时间法更加精准;以相对校正因子法和常规外标法的定量结果显示二者无显著性差异。结论:双标多测法作为一种新的替代对照品方法,能较好地运用于乳块消片多成分的测定中,值得进一步研究完善和推广。
Objective:To establish analysis method of Rukuaixiao tablets for determination of five ingredients(tanshinol,vaccarin,hesperidin,salvianolic acid B and salvianolic acid A) by HPLC,and validate the feasibility of two reference substances for determination of multiple components(TRSDMC)(linear calibration using two reference substances(LCTRS) for qualitative analysis of chromatographic peaks combined with relative correction factor for quantitative analysis). Take the determination of the content of 5 ingredients as example,compare the advantage and disadvantage of both replacement method of chemical reference substance(LCTRS and the qualitative method using relative retention time) and differences between the two quantitative method(using relative correction factor method and external method). Methods:HPLC method was adopted,the mobile phase consisted of acetonitrile(A)-0.2% phosphoric acid aqueous solution(B) with gradient elution(0-30 min,5%A→ 20%A;30-40 min,20%A→ 30%A;40-50 min,30%A) at a flow rate of 1 mL·min-1. The detection wavelength was 282 nm and the column temperature was 30℃. The retention time of the five ingredients in Rukuaixiao tablets was determined on four different C18 columns by HPLC. The standard retention time(SRT) was calculated. Both tanshimol sodium(peak 1) and hesperidin(peak 3) were taken as the linear calibration standards. LCTRS was used for accurately locating the peaks of each ingredient,forecasting the retention time of the other three ingredients when the method was established and validated. Meanwhile the middle peak(hesperidin) was chosen as reference substance for the determination of relative correction factor in order to calculate the content of each component. Results:The content ranges of tanshinol, vaccarin, hesperidin, salvianolic acid B and salvianolic acid in samples from different enterprises were 3.14-6.25 mg·g-1, 0.37-1.79 mg·g-1,0.42-1.18 mg·g-1,0.91-3.50 mg·g-1,1.40-6.97 mg·g-1,respectively. LCTRS could be used for accurately predicting the retention time of the components tested on unknown column and the deviation between the predicted result and the actual value was less than the relative retention time method. LCTRS was more accurate and suitable for chromatographic peak location. It was proved that there’s no difference between the two quantitative methods. Conclusion:LCTRS can be used in the determination of multiple components of Rukuaixiao tablets as a new replacement method of chemical reference substance and is worth further research and improvement.
作者
吴燕红
孙磊
许妍
万林春
王新华
邬秋萍
WU Yan-hong;SUN Lei;XU Yan;WAN Lin-chun;WANG Xin-hua;WU Qiu-ping(Jiangxi Provincial Research Institute for Drug Control,NMPA Key Laboratory of Quality Evaluation of Traditional Chinese Patent Medicine,Jiangxi Provincial Engineering Research Center for Drug and Medical Device Quality,Nanchang 330029,China;National Institutes for Food and Drug Control,Beijing 100050,China;Jiangxi Provincial Medical Product Administration,Nanchang 330029,China)
出处
《药物分析杂志》
CAS
CSCD
北大核心
2020年第10期1790-1798,共9页
Chinese Journal of Pharmaceutical Analysis
基金
江西省食品药品监督管理局科研项目(2016YP07)
国家自然科学基金项目(81303214)。
关键词
双标多测法
双标线性校正法
替代标准物质
相对校正因子
乳块消片
丹参素
王不留行黄酮苷
橙皮苷
丹酚酸
丹参
王不留行
陈皮
two reference substances for determination of multiple components(TRSDMC)
liner calibration with two reference substances(LCTRS)
substitute reference material
relative correction factor
Rukuaixiao tablet
tanshinol
vaccarin
hesperidin
salvianolic acid
Salviae Miltiorrhizae Radix et Rhizoma
Vaccariae semen
Citri Reticulatae Pericarpium
