HPLC法测定山楂炮制前后10种有机酸成分的含量

Content determination of ten organic acids in Crataegus pinnatifida before and after processing by HPLC

关键词:  山楂  HPLC  有机酸  炮制  含量测定

Key words:  Crataegus pinnatifida  HPLC  organic acids  processing  content determination

摘要:

[目的] 建立山楂中10种有机酸成分的高效液相色谱法（HPLC）含量测定方法，探究炮制前后山楂中10种有机酸成分的含量变化规律。[方法] 采用HPLC同时测定草酸、酒石酸、苹果酸、乳酸、柠檬酸、琥珀酸、没食子酸、原儿茶酸、香草酸和咖啡酸这10种有机酸成分的含量，色谱条件为Hypersil ODS2色谱柱（4.6 mm×250 mm，5 μm），流动相：0.1%磷酸水溶液（A）：甲醇（B）梯度洗脱（0～10 min，1%～25% B；10～28 min，25% B；28～30 min，25%～1% B；30～35 min，1% B），检测波长：210 nm，流速：0.8 mL/min，柱温为27.5℃，进样量10 μL。[结果] 10种有机酸在所设浓度范围线性良好，相关系数为0.999 5～0.999 9，精密度RSD均小于1.55%，常温放置24 h的稳定性RSD均小于2.91%，4℃放置24 h的稳定性RSD均小于1.92%，重复性RSD均小于2.47%，加样回收率为95.45%～103.73%，RSD均小于1.83%。[结论] 建立的HPLC测定方法快速高效、重现性好，适合山楂中这10种有机酸成分的含量测定。草酸、酒石酸、苹果酸、乳酸、柠檬酸、没食子酸、香草酸和咖啡酸的含量随炮制温度升高或加热时间延长而下降；琥珀酸的含量先下降后升高；原儿茶酸的含量先升高后下降。

Abstract:

[Objective] To establish an high performance liquid chromatography method (HPLC) method for the determination of 10 organic acids in Crataegus pinnatifida (CP),and to explore the content changes of 10 organic acids in CP before and after processing.[Methods] The contents of 10 organic acids including oxalic acid,tartaric acid,malic acid,lactic acid,citric acid,succinic acid,gallic acid,protocatechuic acid,vanillic acid and caffeic acid were determined by HPLC. The chromatographic conditions were Hypersil ODS2 column (4.6 mm×250 mm,5 μm) and the mobile phase was 0.1% phosphoric acid aqueous solution (A):methanol (B),which was processed as follows:1%~25% B at 0~10 min,25% B at 10~28 min,25%~1% B at 28~30 min,1% B at 30~35 min,detection wavelength:210 nm,flow rate:0.8 mL/min,column temperature:27.5℃,injection volume:10 μL.[Results] The linearity of 10 kinds of organic acids in the set concentration range was good,the correlation coefficient was 0.999 5~0.999 9,the precision RSD was less than 1.55%,the stability RSD was less than 2.91% at room temperature for 24 h,the stability RSD was less than 1.92% at 4℃ for 24 h,the repeatability RSD was less than 2.47%,and the recovery rate was 95.45%~103.73%.[Conclusion] The established HPLC method is rapid,efficient and reproducible,and is suitable for the determination of these 10 organic acids in CP. The contents of oxalic acid,tartaric acid,malic acid,lactic acid,citric acid,gallic acid,vanillic acid and caffeic acid decreased with the increase of processing temperature or heating time. The content of succinic acid decreased first and then increased. The content of protocatechuic acid first increased and then decreased.