桂枝水提液全时段双波长融合高效液相色谱指纹图谱研究及6个成分定量分析

目的： 研究建立桂枝水提液的高效液相色谱双波长融合指纹图谱分析方法，并测定桂枝水提液中原儿茶酸、原儿茶醛、香豆素、肉桂醇、肉桂酸、桂皮醛的含量，为桂枝水提液质量控制提供依据。 方法： 采用Waters Symmetry RP C₁₈色谱柱（4.6 mm×250 mm，5 μm），以乙腈-0.1%磷酸水溶液为流动相，梯度洗脱（0~10 min，3%A→6%A；10~20 min，6%A→10%A；20~35 min，10%A→18%A；35~50 min，18%A→25%A；50~80 min，25%A→30%A；80~90 min，30%A→40%A），流速1.0 mL·min^-1，检测波长254、275 nm，柱温30 ℃。使用Matlab 7.1软件编程，对波长254、275 nm下的CSV格式数据进行全时段双波长融合。 结果： 原儿茶酸、原儿茶醛、香豆素、肉桂醇、肉桂酸、桂皮醛线性范围分别为0.37~14.9、0.05~2.05、0.58~23.37、0.39~15.66、0.67~26.85、2.48~99.23 μg·mL^-1，r为0.9995以上；平均回收率在97.8%~102%，RSD均小于3%（n=6）。双波长融合指纹图谱全面体现了桂枝254 nm、275 nm特征吸收波长的指纹信息，以桂皮醛为参照峰，标定了12个共有峰，指认其中6个峰，分析20批不同产地及批次的桂枝与共有模式之间具有良好的相似性，相似度均在0.90以上。融合后的指纹图谱信息更加丰富，6个物质在20批药材中含量有一定的差异。 结论： 双波长融合技术的高效液相色谱指纹图谱及6个成分的含量测定可以用于桂枝水提物的质量控制。

Objective: To establish the double-wavelength fusion high performance liquid chromatographic fingerprints of water extract of Cinnamomi Ramulus and to determine the contents of protocatechuic acid, protocatechualdehyde, cumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde in the extract, so as to provide a scientific basis for the quality control of Cinnamomi Ramulus water extract. Methods: The column was Waters Symmetry RP C₁₈(4.6 mm×250 mm, 5 μm);The mobile phase, consisting of solvent A(acetonitrile) and solvent B(0.1% aqueous phosphoric acid), was programmed as a linear gradient(0-10 min,3%A→6%A;10-20 min,6%A→10%A;20-35 min,10%A→18%A;35-50 min,18%A→25%A;50-80 min,25%A→30%A;80-90 min,30%A→40%A);The flow rate was 1.0 mL·min^-1;The absorbance was monitored at the wavelengths of 254 and 275 nm ;The column temperature was 30 ℃. Matlab 7.1 was adopted for all-time double-wavelength fusion for the data in CSV format. Results: The calibration curves of protocatechuic acid, protocatechualdehyde, coumarin, cinnamic alcohol, cinnamic acid and cinnamaldehyde were all in good linearity over the ranges of 0.37-14.9, 0.05-2.05, 0.58-23.37, 0.39-15.66, 0.67-26.85, 2.48-99.23 μg·mL^-1(r>0.9995), respectively, and the average recoveries(n=6) ranged from 97.8% to 102.0%, with RSD<3.0%. The results of double-wavelength fingerprint chromatograms showed all-around information of the fingerprints at 254 and 275 nm. The fingerprints containing 12 co-possessing peaks were obtained when choosing cinnamaldehyde peak as the reference peak. The similarity among 20 batches of Cinnamomi Ramulus was over 0.90;The information of the fusion fingerprints was more abundant. The contents of the six compounds were different in the 20 batches of Cinnamomi Ramulus. Conclusion: The systematic quantified fingerprint method based on the technique of double-wavelength fusion fingerprint and the determination of the six constituents can be used for the quality control of the water extract of Cinnamomi Ramulus.