关于《中国药典》2020年版红花山柰酚含量测定的探讨

Discussion on determination of kaempferol in safflower in the Chinese pharmacopoeia 2020

目的:采用《中国药典》2020年版测定红花项下山柰酚的含量时,6-甲氧基山柰酚完全包埋于山柰酚的色谱峰中,形成共流物出峰,导致测量结果偏高,为准确测定红花的含量,建立了红花药材中山柰酚和6-甲氧基山柰酚的含量测定方法,并测定了30批不同产地红花药材含量。方法:采用高效液相色谱法,以waters Symmetry Shield RP 18(4.6 mm×250 mm,5μm)为色谱柱,以甲醇-0.4%磷酸溶液(70∶30)为流动相,柱温为25℃,流速为1.0 mL•min^(-1),进样量为10μL,检测波长为367 nm。结果:山柰酚在0.00472~0.472μg(r=0.999)、6-甲氧基山柰酚在0.00456~0.456μg(r=0.999)范围内呈良好线性关系。山柰酚、6-甲氧基山柰酚的平均回收率分别为98.2%、99.8%,测定了30批红花药材的山柰酚的和6-甲氧基山柰酚的含量范围分别为0.026%~0.071%、0.007%~0.052%。结论:所建立的方法解决了原方法测定定量不准确的问题,建议《中国药典》2020年版红花药材增加6-甲氧基山柰酚含量测定指标并进行单指标限度控制:以干燥品计,含山柰酚不得少于0.034%,6-甲氧基山柰酚不得少于0.016%。

Objective:When the content of kaempferol in safflower was determined by the Chinese pharmacopoeia 2020,6-methoxykaempferol was completely embedded in the chromatographic peak of kaempferol,which resulted in the formation of co-flow peak,resulting in higher measurement results,to determine the content of kaempferol and 6-methoxykaempferol in safflower,a method for the determination of kaempferol and 6-methoxykaempferol in safflower was established,The contents of 30 batches of safflower from different producing areas were determined.Methods:Waters Symmetry Shield RP 18 column(4.6 mm×250 mm,5μm)was used with methanol-0.4%phosphoric acid(70∶30)as mobile phase.The column temperature was 25℃,the flow rate was 1.0 mL•min^(-1),the injection volume was 10μL,and the detection wavelength was 367 nm.Results:Kaempferol showed a good linear relationship in the range of 0.00472-0.472μg(r=0.999)and 6-methoxykaempferol in the range of 0.00456-0.456μg(r=0.999).The average recoveries of kaempferol and 6-methoxykaempferol were 98.2%and 99.8%,respectively,the content ranges of kaempferol and 6-methoxykaempferol in 30 batches of Safflower were 0.026%-0.071%and 0.007%-0.052%,respectively.Conclusions:The established method solved the problem of inaccuracy in the determination of 6-methoxykaempferol by the original method.It is suggested that the content of 6-methoxykaempferol should be increased in the Chinese Pharmacopoeia 2020 and the limit control of single index should be carried out.The content of 6-methoxykaempferol should not be less than 0.034%and that of 6-methoxykaempferol should not be less than 0.016%.