Wang Tao, Zhang Hui, Zhao Dan, Chen Jie, Xie Zhuo-Lin, Dai Wei, Ye Kun-Hao, Huang Jing, Chen Gang-Fu
Institute of Chinese Medicinal Materials,Mianyang Academy of Agricultural Sciences Mianyang 621023,China Comprehensive Experimental Station of Mianyang,Chinese Materia Medica of China Agriculture Research System Mianyang 621023,China.
Biotechnology Center,Mianyang Academy of Agricultural Sciences Mianyang 621023,China.
Zhongguo Zhong Yao Za Zhi. 2019 Jan;44(2):338-343. doi: 10.19540/j.cnki.cjcmm.20181009.011.
This project was launched to and establish the wavelength overlapping HPLC fingerprint for Danshen and determine the contents of 7 component in Danshen. The chromatographic fingerprints were built by using Agilent Eclipse Plus C_(18)( 4. 6 mm×100 mm,3. 5 μm) as stationary phase and 0. 1% formic acid solution( A)-acetonitrile( B) as mobile phase with gradient elution( 0-5 min,10%-20% B; 5-20 min,20%-30% B; 20-25 min,30%-50% B; 25-40 min,50%-65% B; 40-45 min,65%-80% B; 45-46 min,80%-10% B; 46-50 min,10% B) at a flow rate of 1 mL·min~(-1). The column temperature was maintained at 30 ℃ and the detection wavelength was set at 250,280,310 and 340 nm. The technique of wavelength overlapping fingerprint was established and the contents of 7 indicative compounds have been determined in this method. The results of wavelength overlapping HPLC fingerprint showed all-around information of the fingerprints at 250,280,310 and 340 nm,and the similarity among 17 batches of Danshen was over 0. 828-0. 936. In wavelength overlapping HPLC fingerprint,15 common peaks were selected as the common peaks,and 7 contents of them were indentified as rosmarinic acid,salvianolic acid B,salvianolic acid A,phenolic acid,dihydrotanshinone Ⅰ,tanshinone Ⅰ and tanshinone ⅡA. The results of methodological study demonstrated that the method met the requirements of the determination. The method established in this study is simple,convenient and durable,which can provide a scientific basis for the quality control of Danshen.
本项目旨在建立丹参的波长重叠高效液相色谱指纹图谱并测定丹参中7种成分的含量。采用安捷伦Eclipse Plus C₁₈(4.6 mm×100 mm,3.5 μm)为固定相,以0.1%甲酸溶液(A)-乙腈(B)为流动相进行梯度洗脱(0 - 5分钟,10% - 20%B;5 - 20分钟,20% - 30%B;20 - 25分钟,30% - 50%B;25 - 40分钟,50% - 65%B;40 - 45分钟,65% - 80%B;45 - 46分钟,80% - 10%B;46 - 50分钟,10%B),流速为1 mL·min⁻¹,建立色谱指纹图谱。柱温保持在30℃,检测波长设定为250、280、310和340 nm。建立了波长重叠指纹图谱技术,并采用该方法测定了7种指标性成分的含量。波长重叠高效液相色谱指纹图谱结果显示,在250、280、310和340 nm处的指纹图谱信息全面,17批丹参之间的相似度在0.828 - 0.936之间。在波长重叠高效液相色谱指纹图谱中,选取15个共有峰作为参照峰,其中7个峰的成分被鉴定为迷迭香酸、丹酚酸B、丹酚酸A、酚酸、二氢丹参酮Ⅰ、丹参酮Ⅰ和丹参酮ⅡA。方法学研究结果表明该方法符合测定要求。本研究建立的方法简便、耐用,可为丹参的质量控制提供科学依据。