1.河南中医药大学 药学院,河南省中药质量控制与评价工程技术研究中心,郑州 450046
2.漯河医学高等专科学校,河南 漯河 462002
3.河南中医药大学 第三附属医院,郑州 450046
段浩瀚,在读硕士,从事中药质量分析研究,E-mail:2978946882@qq.com
雷敬卫,博士,教授,从事中药质量分析研究,Tel:0371-656955281,E-mail:925390812@qq.com
收稿:2023-05-19,
网络出版:2023-07-27,
纸质出版:2024-03-05
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段浩瀚,贾豪,雷敬卫等.基于颜色-成分关联分析比较不同产地牛膝炮制过程中质量变化的规律[J].中国实验方剂学杂志,2024,30(05):143-151.
DUAN Haohan,JIA Hao,LEI Jingwei,et al.Comparison of Quality Change During Processing Process of Achyranthis Bidentatae Radix from Different Origins Based on Color-component Correlation Analysis[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(05):143-151.
段浩瀚,贾豪,雷敬卫等.基于颜色-成分关联分析比较不同产地牛膝炮制过程中质量变化的规律[J].中国实验方剂学杂志,2024,30(05):143-151. DOI: 10.13422/j.cnki.syfjx.20231263.
DUAN Haohan,JIA Hao,LEI Jingwei,et al.Comparison of Quality Change During Processing Process of Achyranthis Bidentatae Radix from Different Origins Based on Color-component Correlation Analysis[J].Chinese Journal of Experimental Traditional Medical Formulae,2024,30(05):143-151. DOI: 10.13422/j.cnki.syfjx.20231263.
目的
2
选取不同产地牛膝作为样品,对不同炮制程度样品的化学成分及色度进行定量分析,明确牛膝炮制过程中色度与成分的关联性与变化规律,以期为牛膝炮制品质量评价提供参考。
方法
2
使用色差仪测量不同产地牛膝炮制不及、炮制适中、炮制过度3种炮制程度的色度值,显示牛膝炮制过程中色值变化趋势,借助主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)等分析方法对不同炮制程度酒牛膝与盐牛膝样品的颜色参数进行分析。使用高效液相色谱法(HPLC)测定牛膝的8种代表性成分含量,借助皮尔逊相关性分析对色度与各代表性成分之间的相关性进行分析,使用Fisher线性判别分析进一步验证所选的8个代表成分的适用性,对不同炮制程度的酒牛膝和盐牛膝按照炮制程度进行分组建模,不同炮制程度样品各以48个样本作为训练样本,以5-羟甲基糠醛、水龙骨素B、
β
-蜕皮甾酮、25
R
-牛膝甾酮、25
S
-牛膝甾酮、人参皂苷Ro、竹节参皂苷Ⅳa、多糖含量为变量,2种炮制品分别建立一个判别函数,将不同炮制程度的酒牛膝和盐牛膝各12个样本进行回代对判别函数进行验证,检验函数可靠性。
结果
2
PCA、OPLS-DA结果显示,不同炮制程度牛膝各自归类成簇,结果能够将不同炮制品显著区分。牛膝酒炙和盐炙过程中5-羟甲基糠醛、人参皂苷Ro、竹节参皂苷Ⅳa随着炮制程度的加深含量逐渐升高,水龙骨素B、
β
-蜕皮甾酮、25
R
-牛膝甾酮、25
S
-牛膝甾酮和总多糖含量呈现逐渐降低趋势,说明酒炙和盐炙过程中8个成分升高与降低的程度不同。皮尔逊相关性分析结果显示,不同炮制程度酒牛膝和盐牛膝样品的5-羟甲基糠醛含量与明度值(
L
*
)、总色差值(
E
*
ab
)呈负相关(
P
<
0.01),与红绿值(
a
*
)、黄蓝值(
b
*
)呈正相关(
P
<
0.01);水龙骨素B、多糖含量与
L
*
、
E
*
ab
呈正相关(
P
<
0.01)。使用Fisher线性判别分析建立酒牛膝和盐牛膝的不同炮制程度的判别函数,训练样本酒牛膝判别函数正确率为93.75%,盐牛膝判别函数正确率为95.83%。将12个不同炮制程度酒牛膝测试样本和12个不同炮制程度盐牛膝测试样本进行回代,正确率均为100%。
结论
2
不同产地不同炮制程度的牛膝成分和颜色变化趋势较为一致,通过色值可将不同炮制程度牛膝较好区分,牛膝在炮制过程中颜色与部分代表性成分具有相关性,说明色度可为牛膝炮制程度鉴别和成分含量预测提供参考。
Objective
2
Taking Achyranthis Bidentatae Radix(ABR) from different origins as samples, to quantitatively analyze the chemical composition and chromaticity of ABR with different processing degrees, and clarify the correlation and change law between color and composition in the processing process of ABR, so as to provide reference for the quality evaluation of processed products of ABR.
Method
2
The colorimeter is used to measure the chromaticity values of three kinds of processing degrees of ABR in different origins to show the color value change trend during the processing process, and the color parameters of wine-processed and salt-processed products of ABR with different processing degrees were analyzed by principal component analysis(PCA), orthogonal partial least squares-discriminant analysis(OPLS-DA) and other analysis methods. The contents of eight representative components of ABR were measured by high performance liquid chromatography(HPLC), the correlation between chromaticity and each representative component was analyzed by Pearson correlation analysis, and the applicability of the selected eight representative components was further verified by Fisher linear discriminant analysis, and the wine-processed and salt-processed products of ABR with different processing degrees were grouped according to the degree of processing, and 48 samples of wine-processed and salt-processed products with different processing degrees were used as training samples. Taking the contents of 5-hydroxymethylfurfural, polypodine B,
β
-ecdysterone, 25
R
-inokosterone, 25
S
-inokosterone, ginsenoside Ro, chikusetsusaponin Ⅳa and polysaccharides as variables, the discriminant function was established respectively, and 12 samples of wine-processed and salt-processed products of ABR with different processing degrees were back-tested to verify the discriminant function and test the reliability of the function.
Result
2
PCA and OPLS-DA results showed that ABR samples with different processing degrees were classified into clusters, and the results could significantly distinguish different processed products. During the process of wine and salt processing, the contents of 5-hydroxymethylfurfural, ginsenoside Ro
,
and chikusetsusaponin Ⅳa gradually increased with the deepening of the processing degree, while the contents of polypodine B,
β
-ecdysterone, 25
R
-inokosterone, 25
S
-inokosterone and polysaccharides showed a gradual decreasing trend, indicating these 8 components increased and decreased to different degrees in the process of wine and salt processing. The results of Pearson correlation analysis showed that the 5-hydroxymethylfurfural content of the samples with different processing degrees of wine-processed and salt-processed products were negatively correlated with the brightness value(
L
*
) and the total color difference value(
E
*
ab
)(
P
<
0.01), and positively correlated with the red-green value(
a
*
) and the yellow-blue value(
b
*
)(
P
<
0.01), and that the content of polypodine B and polysaccharides were positively correlated with
L
*
and
E
*
ab
(
P
<
0.01). The discriminant functions of wine-processed and salt-processed products of ABR were established by Fisher linear discriminant analysis, and their accuracy rates in the training samples were 93.75% and 95.83%, respectively. Twelve test samples of wine-processed and salt-processed products with different processing degree were back substitution, and the correct rate was 100%.
Conclusion
2
The trend of composition and color changes of ABR with different processing degrees in different production areas is relatively consistent, and the color value can better distinguish ABR with different processing degrees, and the color of ABR is related to some representative components in the processing process, indicating that the color can provide reference for the identification of the processing degree of ABR and the prediction of component content.
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