摘要
In this paper, we proposed a novel resolution criterion(improved calibrated normalized resolution product, r*') to evaluate separation quality of fingerprints. By comparing with the calibrated normalized resolution product(r*) and the hierarchical chromatographic response function(HCRF), the validity of this criterion was demonstrated by experimental chromatograms. The soy isoflavone extract was selected as the analytical object. The initial and end percentages of methanol and elution time affecting gradient elution were tested by orthogonal design. The final optimized conditions were as follows. It was detected by UV absorbance at 254 nm, column temperature was maintained at 36 oC, solvent A was 0.1%(v/v) acetic acid, solvent B was methanol, gradient elution was from 34% to 65% B in a linear gradient in 25 min, and the flow-rate was set at 1.0 m L/min. In addition, the main ingredients of the soy isoflavone extract were confirmed by LC-ESI/MS.
本文提出了一种新的评价色谱指纹图谱分离质量的指标,即改进的校正标准分离度积(ri*')。以大豆异黄酮提取物为分析对象,通过正交设计对高效液相梯度洗脱的初始甲醇比例、洗脱结束时的甲醇比例以及洗脱时间进行考察,用改进的校正标准分离度积(ri*')衡量所得色谱图,通过与校正标准分离度积(ri*)及谱系色谱响应函数(HCRF)进行比较,分析各个指标的优缺点,论证了改进的校正标准分离度积(ri*')这一评估指标的有效性,并建立了大豆异黄酮提取物指纹图谱的优化色谱条件(流动相A相:0.1%醋酸,B相:甲醇;梯度洗脱B相:;流速:1.0 m L/min检测波长:254 nm;柱温:36 oC)。此外,用高效液相色谱-电喷雾质谱法对大豆异黄酮提取物的活性成分进行了归属分析。25 min34%????65%
基金
National Higher-Education Institution General Research and Development Funding(Grant No.JKP2011010)