运用近红外光谱技术快速测定大青叶中靛玉红含量

Rapid Determination of Indirubin Content in Isatidis Folium Using Near-infrared Spectroscopy

目的运用近红外光谱技术快速测定大青叶中靛玉红含量,建立靛玉红的近红外定量分析模型,并通过模型分析评价大青叶的品质。方法采用传统的高效液相色谱法对74批大青叶中靛玉红含量进行测定及方法学验证,运用偏最小二乘法建立靛玉红含量与近红外光谱之间的定量校正分析模型,并对验证集样品进行含量预测,将预测值与实际测定值进行统计分析。结果靛玉红浓度在1.04~3.13μg·ml^(-1)范围内线性良好,回归方程为Y=11566X+911.7,r=0.9990;精密度、重复性、12 h内稳定性的RSD分别为0.31%、0.86%、1.11%;加样平均回收率为99.22%。建立的定量分析模型准确性好,内部交叉验证均方差、内部交叉验证决定系数分别为0.226、0.983。通过对24份校正集样品进行外部验证,近红外预测值与高效液相测定值的预测回收率为93.66%。对两组数据进行方差显著性分析,其显著性指标值分别为0.540和0.428,表明两种试验方法的数据差异无统计学意义,该模型的精度和再现性均符合规定。结论近红外光谱技术对大青叶中靛玉红含量测定结果较好,建立的定量分析模型可以无损、快速、准确地对大青叶进行质量评价,提高生产效率。

Objective To determine the content of indirubin in Folium Isatidis using near-infrared spectroscopy,establish a near-infrared quantitative analysis model for indirubin,and to evaluate the quality of Isatidis Folium.Methods The contents of indirubin in 74 batches of Isatidis Folium were determined using high-performance liquid chromatography before being methodologically validated.A quantitative calibration analysis model between indirubin contents and near-infrared spectra was established by using partial least squares.The content prediction was performed using the samples for verification,and the predicted values were statistically analyzed along with the values actually measured.Results The linear range of indirubin concentrations was 1.04-3.13μg·ml^(-1),the regression equation was Y=11566X+911.7,r=0.9990,the precision RSD was 0.31%,the repeatability RSD was 0.86%,the stability was good within 12 hours,the RSD was 1.11%and the average recovery was 99.22%.This quantitative analysis model was quite accurate,with an RMSECV(internal cross-validation mean square deviation)and R2(internal cross-validation decision coefficient)of 0.226 and 0.983,respectively.The predicted recoveries of nearinfrared spectroscopy and high-performance liquid chromatography were 93.66%based on external validation of 24 calibration samples.The two groups of data were analyzed for the significance of variance,and the significance index values were 0.540 and 0.428,respectively.The precision and reproducibility of the model met the requirements.Conclusion Near-infrared spectroscopy can accurately predict indirubin contents in Isatidis Folium,and the established quantitative analysis model can be used to evaluate the quality of Isatidis Folium quickly and accurately while improving productivity.