熟地黄蒸制过程特征图谱和颜色变化研究及炮制工艺优选

刘燎原; 纪玉华; 王文丽; 甘力帆; 黄瑶; 曾昭君; 张志鹏

doi:10.14148/j.issn.1672-0482.2023.0764

熟地黄蒸制过程特征图谱和颜色变化研究及炮制工艺优选

doi: 10.14148/j.issn.1672-0482.2023.0764

广东一方制药有限公司，广东省中药配方颗粒企业重点实验室, 广东佛山 528244

基金项目:

中华人民共和国工业和信息化部2022年产业技术基础公共服务平台项目 2022-230-221

详细信息

作者简介:
刘燎原, 男, 副主任中药师, E-mail: 99263@163.com

通讯作者:
张志鹏, 男, 副主任中药师, 主要从事中药饮片及配方颗粒质量标准研究, E-mail: zhangzp0909@163.com

中图分类号: R284
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- 文章访问数: 130
- HTML全文浏览量: 14
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- 被引次数: 0
出版历程
- 收稿日期: 2023-03-28
- 网络出版日期: 2023-08-17
- 发布日期: 2023-08-10

Study on the Characteristic Specturm and Color Change of Steamed Rehmanniae Radix Praeparata and Optimization of Processing Technology

Guangdong Yifang Pharmaceutical Co. Ltd., Guangdong Province Key Laboratory of Traditional Chinese Medicine Formula Granule, Foshan 528244, China

摘要

摘要: 目的研究熟地黄蒸制过程中特征图谱与颜色的变化规律, 结合Box-Behnken响应曲面模型, 优选熟地黄炮制工艺。方法应用Pearson相关性分析、聚类分析、主成分分析(PCA)、正交偏最小二乘分析(OPLS-DA)等多元统计方法研究熟地黄蒸制过程中的颜色和特征图谱峰面积变化, 筛选熟地黄药材关键的质量标志物(Q-markers), 以Q-markers为评价指标, 结合综合评分法考察蒸制时间、干燥温度、干燥时间对熟地黄质量的影响, 同时采用Box-Behnken响应曲面法优选最佳炮制工艺。结果建立的生地黄特征图谱和熟地黄特征图谱分别标定18、20个共有峰, 其中19号峰[5-羟甲基糠醛(5-HMF)]为生地黄经炮制后产生的新化学成分; 生地黄蒸制2 h后色差显著(ΔE^*>12), 且各特征峰峰面积与生品的差异较大; 蒸制时间超过2 h后, 不同蒸制时间点的熟地黄ΔE^*为0.28~4.76, 色差不能被肉眼识别。以色度值L^*、a^*、b^*、E^*为变量可将不同炮制点的熟地黄聚为3类, 以色度值L^*、a^*、b^*、E^*, 环烯醚萜苷类成分, 苯乙醇苷类成分, 5-HMF为变量可将不同炮制点的熟地黄聚为4类。熟地黄最佳炮制工艺为: 润制水量为药材量的0.3倍, 润制时间为24 h, 蒸制时间为2.17 h, 干燥温度为61.15 ℃, 干燥时间为13.73 h。结论建立的响应面模型准确、可预测, 建立的特征图谱方法和优选的熟地黄炮制工艺方法稳定可行, 结合炮制过程的颜色变化规律, 可为熟地黄的质量控制和综合开发利用提供参考。
- 地黄 /
- 熟地黄 /
- 特征图谱 /
- 炮制 /
- 色度值 /
- 响应曲面法
Abstract: OBJECTIVE To optimize the processing technique of Shudihuang (Rehmanniae Radix Praeparata, RRP) by Box-Behnken response surface model based on the change rule of characteristic spectrum and color. METHODS Multivariate statistical methods including Pearson correlation analysis, cluster analysis, principal component analysis (PCA) and orthogonal partial least squares-discriminate analysis (OPLS-DA) were used to analyze the color changes and the peak area changes of the characteristic spectrum in the processing of RRP, and to screen key quality markers (Q-markers) for RRP. Q-markers were used to evaluate the effects of steaming time, drying temperature, and drying time on the quality of RRP, and the Box-Behnken response surface method was used to determine the optimal processing technique. RESULTS The established characteristic spectrum of Shengdihuang (Rehmanniae Radix, RR) and RRP marked 18 and 20 common peaks, respectively. Among them, peak 19(5-HMF)was a new chemical component produced after the processing of RR. There was a significant difference in color (ΔE^* > 12) after 2 h of evaporation, and the peak area of each characteristic was significantly different from RR. The ΔE^* of RRP at different steaming time points was 0.28~4.76, indicating that the color difference could not be recognized by the naked eye. RRP from different processing points could be clustered into 3 groups with chromaticity values L^*, a^*, b^*, E^* as variables, and RRP from different processing points could be clustered into 4 groups with chromaticity values L^*, a^*, b^*, E^*, and iridoid glycosides, phenylethanolglycosides and 5-HMF as variables. The optimum processing technique of steamed RRP was as follows: moistening water was 0.3 times the amount of medicinal material, moistening time was 24 h, steaming time was 2.17 h, drying temperature was 61.15 ℃, and drying time was 13.73 h. CONCLUSION The established response surface model is accurate and predictable, the established characteristic spectrum and the optimized processing method of RRP are stable and feasible. Combining with the color change rule of the processing process, it can provide a reference for comprehensive utilization and the quality control of RRP.
- Rehmanniae Radix /
- Rehmanniae Radix Praeparata /
- characteristic spectrum /
- processing /
- chromaticity value /
- response surface method

HTML全文

图 1 14批生地黄(A)、熟地黄(B)的叠加特征图谱及其对照特征图谱(R)、混合对照品溶液的UPLC图(C~D)

注: 3.腺苷; 4.梓醇; 7.地黄苷D; 8.地黄苷A; 9.益母草苷; 11.洋地黄叶苷C; 12.焦地黄苯乙醇苷A1;13.毛蕊花糖苷; 14.焦地黄苯乙醇苷B1;15.异毛蕊花糖苷; 19.5-HMF

Figure 1. UPLC superimposed characteristic spectrum and control characteristic spectrum (R) of 14 batches of RR (A) and RRP (B), chromatograms of mixed reference solution (C-D)

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图 2 聚类分析图

Figure 2. Cluster analysis diagram

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图 3 熟地黄炮制过程色度值PCA二维载荷图(A)、PCA得分图(B)、OPLS-DA得分图(C)、VIP得分图(D)

Figure 3. PCA two-dimensional loading diagram (A), PCA score chart (B), OPLS-DA score chart (C), VIP score chart (D) of chromaticity value of processed RRP

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图 4 熟地黄炮制过程色度值和化学成分PCA二维载荷图(A)、PCA得分图(B)、OPLS-DA得分图(C)、VIP得分图(D)

Figure 4. PCA two-dimensional loading diagram (A), PCA score chart (B), OPLS-DA score chart (C), VIP score chart (D) of chromaticity value and chemical composition of processed RRP

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图 5 因素A、B、C的交互作用3D图

Figure 5. 3D diagram of the interaction of factors A, B and C

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表 1 炮制过程熟地黄色度值与色差值

Table 1. Chromaticity value and color difference value of RRP during processing

编号	蒸制时间/h	L^*	a^*	b^*	E^*	ΔL^*	Δa^*	Δb^*	ΔE^*
PZ1	0	17.37	8.78	14.06	24.01	/	/	/	/
PZ2	2	16.14	1.76	-2.54	16.43	-1.23	-7.02	-16.60	18.06
PZ3	4	15.69	0.57	0.29	15.70	-0.45	-1.20	2.83	3.11
PZ4	6	12.50	2.13	-2.88	13.00	-3.19	1.56	-3.18	4.76
PZ5	8	13.19	1.47	-3.57	13.75	0.69	-0.67	-0.69	1.18
PZ6	12	12.92	1.98	-3.68	13.58	-0.27	0.52	-0.12	0.60
PZ7	18	14.17	1.95	-3.39	14.70	1.24	-0.03	0.29	1.28
PZ8	24	14.12	2.18	-3.25	14.65	-0.04	0.23	0.14	0.28
PZ9	30	13.21	2.06	-3.20	13.74	-0.92	-0.12	0.05	0.93
PZ10	36	13.57	2.00	-2.91	14.03	0.37	-0.06	0.29	0.47
PZ11	42	14.30	2.10	-2.58	14.68	0.73	0.10	0.34	0.81
PZ12	48	13.06	2.75	-2.80	13.64	-1.24	0.65	-0.23	1.42

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表 2 炮制过程熟地黄色度值与特征图谱各特征峰峰面积Pearson相关系数

Table 2. Pearson correlation coefficient between the chromaticity value of RRP and the peak area of each characteristic spectrum in the processing process

峰号	L^*	a^*	b^*	E^*
1	0.546	0.316	0.360	0.490
2	0.324	0.604^*	0.659^*	0.564
3	0.135	-0.344	-0.175	-0.094
4	0.678^*	0.632^*	0.821^**	0.791^**
5	0.786^**	0.827^**	0.924^**	0.938^**
6	-0.413	-0.162	-0.355	-0.390
7	0.609^*	0.580^*	0.745^**	0.715^**
8	0.646^*	0.633^*	0.749^**	0.753^**
9	0.207	0.182	0.163	0.249
10	0.725^**	0.872^**	0.960^**	0.921^**
11	0.008	-0.329	-0.154	-0.143
12	0.465	-0.075	0.321	0.287
13	0.297	-0.143	0.213	0.161
14	0.496	0.097	0.435	0.409
15	0.723^**	0.539	0.757^**	0.755^**
16	-0.307	-0.453	-0.399	-0.390
17	-0.375	-0.845^**	-0.620^*	-0.658^*
18	0.174	-0.226	0.123	0.031
19	-0.123	-0.453	-0.216	-0.281
20	0.459	0.051	0.361	0.351
4+7+8+9	0.685^*	0.652^*	0.831^**	0.804^**
11+12+13+14+15	0.425	-0.026	0.329	0.298
注: ^P < 0.05, ^{ *}P < 0.01。其中特征图谱4+7+8+9为环烯醚萜苷类成分总峰面积, 11+12+13+14+15为苯乙醇苷类成分总峰面积。

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表 3 响应曲面试验方案和结果

Table 3. Response surface test scheme and results

序号	A/h	B/℃	C/h	环烯醚萜苷类成分总峰面积	苯乙醇苷类成分总峰面积	5-HMF峰面积	Z
1	4	80	12	584 925	217 776	43 515	0.64
2	4	60	20	762 637	282 999	52 970	0.83
3	2	70	12	870 639	278 986	33 510	0.89
4	4	70	16	683 565	268 804	56 931	0.77
5	2	80	16	640 467	217 733	61 293	0.69
6	4	70	16	714 345	245 392	55 595	0.76
7	4	70	16	651 813	249 422	54 340	0.72
8	2	60	16	867 040	306 427	31 742	0.91
9	4	70	16	676 858	276 198	55 434	0.77
10	6	60	16	680 852	257 660	124 773	0.79
11	6	80	16	377 162	131 480	166 302	0.49
12	4	60	12	674 540	308 428	49 173	0.79
13	6	70	12	394 699	223 488	135 284	0.57
14	4	80	20	381 728	197 189	98 335	0.51
15	4	70	16	671 958	256 996	58 586	0.75
16	2	70	20	627 556	245 800	40 741	0.70
17	6	70	20	597 904	194 399	140 905	0.69

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表 4 回归模型方差分析结果

Table 4. Regression model variance analysis results

来源	平方和	自由度	离差平方和	F值	P值	来源	平方和	自由度	离差平方和	F值	P值
模型	0.22	9	0.024	81.81	< 0.000 1	A²	2.063×10^-4	1	2.063×10^-4	0.69	0.433 8
A	0.053	1	0.053	176.46	< 0.000 1	B²	3.069×10^-3	1	3.069×10^-3	10.26	0.015 0
B	0.12	1	0.12	409.35	< 0.000 1	C²	5.012×10^-3	1	5.012×10^-3	16.75	0.004 6
C	3.200×10^-3	1	3.200×10^-3	10.69	0.013 7	残差	2.095×10^-3	7	2.993×10^-4
AB	1.600×10^-3	1	1.600×10^-3	5.35	0.054 0	失拟项	3.750×10^-4	3	1.250×10^-4	0.29	0.831 0
AC	0.024	1	0.024	80.27	< 0.000 1	纯误差	1.720×10^-3	4	4.300×10^-4
BC	7.225×10^-3	1	7.225×10^-3	24.14	0.001 7	总和	0.22	16

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