HPLC指纹图谱结合正交试验优化红参的炮制工艺(1)
中圖分类号 R284.1 文献标志码 A 文章编号 1001-0408(2020)10-1228-06
DOI 10.6039/j.issn.1001-0408.2020.10.14
摘 要 目的：建立红参的高效液相色谱(HPLC)指纹图谱，并优选其最优炮制工艺。方法：采用HPLC法。色谱柱为Waters SymmetryShieldTM RP18，流动相为乙腈-水(梯度洗脱)，柱温为30 ℃，流速为1.0 mL/min，检测波长为203 nm，进样量为10 μL。以人参皂苷Rb1为参照，绘制10批红参样品的HPLC指纹图谱;采用《中药色谱指纹图谱相似度评价系统(2012A版)》进行相似度评价，并确定共有峰。以蒸制温度、蒸制时间、干燥方法为考察因素，人皂苷类成分含量、指纹图谱相似度为指标，采用L16(43)正交试验优化红参的炮制工艺并进行验证;采用SPSS 19.0软件对10批红参样品和最优工艺炮制品进行聚类分析。结果：10批红参共有13个共有峰，相似度均大于0.920;共指认人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1等3个共有峰。最优炮制工艺为100 ℃蒸制150 min，60 ℃干燥;验证试验结果显示，3批红参最优工艺炮制品中人参皂苷Rg1、Re、Rb1的含量分别为0.26%～0.29%、0.17%～0.20%、0.47%～0.54%，其指纹图谱与对照图谱相似度均大于0.970。聚类分析结果显示，10批红参及3批最优工艺炮制品可聚为两类，即HS3～HS10聚为一类、3批最优工艺炮制品及HS1、HS2聚为一类。结论：所建指纹图谱可用于红参的炮制工艺优化，能表征炮制工艺参数波动与药材整体质量的相关性变化;所得最优炮制工艺合理可行。
关键词 红参;炮制工艺优化;正交试验;高效液相色谱法;指纹图谱;聚类分析
Optimization of Processing Technology of Ginseng Radix et Rhizoma Rubra by HPLC Fingerprint Combined with Orthogonal Test
HOU Xinlian1，HUANG Lu2，PENG Cheng2，ZHU Yaning1，CAI Bangjun1，ZHOU Qinmei2，3 [1. China Resources Sanjiu(Ya’an) Pharmaceutical Co.， Ltd.， Sichuan Ya’an 625000， China; 2. Pharmacy College， Chengdu University of TCM， Chengdu 611137， China; 3. Institute of Innovative Medicine Ingredients of Southwest Specialty Medicinal Materials， Chengdu 611137， China]
ABSTRACT OBJECTIVE： To establish an HPLC fingerprint of Ginseng Radix et Rhizoma Rubra， and to optimize its processing technology. METHODS： HPLC method was adopted. The determination was performed on Waters SymmetryShieldTM RP18 column with mobile phase consisted of acetonitrile-water (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃， and the detection wavelength was 203 nm. The sample size was 10 μL. Using ginsenoside Rb1 as reference peak， HPLC fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra was established. The similarity of them was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint (2012 A edition) to confirm common peak. With steaming temperature， time and drying method as factors， using the content of ginsenoside and fingerprint similarity as index， the processing technology was optimized with L16(43) orthogonal test design and verified. Cluster analysis was conducted with SPSS 19.0 statistical software of 10 batches of Ginseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample. RESULTS： There were a total of 13 common peaks in the fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra. The similarity was more than 0.920; 3 common peaks were identified， such as ginsenoside Rg1， ginsenoside Re， ginsenoside Rb1. The optimal processing technology included that steamed at 100 ℃ for 150 min， dried at 60 ℃. The results of validation test show that the contents of ginsenoside Rg1， Re and Rb1 were 0.26%-0.29%， 0.17%-0.20%， 0.47%-0.54%， and the similarity between 3 batches of Ginseng Radix et Rhizome Rubra optimal processed sample and the control fingerprints was more than 0.970. The results of cluster analysis showed that 10 batches of Gimseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample could be clustered into two categories; HS3-HS10 could be clustered into one category， and 3 batches of optimal processed sample， HS1 and HS2 be clustered into one category. CONCLUSIONS： Established fingerprint can be used for the optimization of processing technology of Gimseng Radix et Rhizoma Rubra， and characterize the correlation between fluctuation of technology parameter and quality of medicinal material; the optimal processing technology is reasonable and feasible., 百拇医药(侯新莲 黄露 彭成 朱雅宁 蔡帮军 周勤梅)
DOI 10.6039/j.issn.1001-0408.2020.10.14
摘 要 目的：建立红参的高效液相色谱(HPLC)指纹图谱，并优选其最优炮制工艺。方法：采用HPLC法。色谱柱为Waters SymmetryShieldTM RP18，流动相为乙腈-水(梯度洗脱)，柱温为30 ℃，流速为1.0 mL/min，检测波长为203 nm，进样量为10 μL。以人参皂苷Rb1为参照，绘制10批红参样品的HPLC指纹图谱;采用《中药色谱指纹图谱相似度评价系统(2012A版)》进行相似度评价，并确定共有峰。以蒸制温度、蒸制时间、干燥方法为考察因素，人皂苷类成分含量、指纹图谱相似度为指标，采用L16(43)正交试验优化红参的炮制工艺并进行验证;采用SPSS 19.0软件对10批红参样品和最优工艺炮制品进行聚类分析。结果：10批红参共有13个共有峰，相似度均大于0.920;共指认人参皂苷Rg1、人参皂苷Re、人参皂苷Rb1等3个共有峰。最优炮制工艺为100 ℃蒸制150 min，60 ℃干燥;验证试验结果显示，3批红参最优工艺炮制品中人参皂苷Rg1、Re、Rb1的含量分别为0.26%～0.29%、0.17%～0.20%、0.47%～0.54%，其指纹图谱与对照图谱相似度均大于0.970。聚类分析结果显示，10批红参及3批最优工艺炮制品可聚为两类，即HS3～HS10聚为一类、3批最优工艺炮制品及HS1、HS2聚为一类。结论：所建指纹图谱可用于红参的炮制工艺优化，能表征炮制工艺参数波动与药材整体质量的相关性变化;所得最优炮制工艺合理可行。
关键词 红参;炮制工艺优化;正交试验;高效液相色谱法;指纹图谱;聚类分析
Optimization of Processing Technology of Ginseng Radix et Rhizoma Rubra by HPLC Fingerprint Combined with Orthogonal Test
HOU Xinlian1，HUANG Lu2，PENG Cheng2，ZHU Yaning1，CAI Bangjun1，ZHOU Qinmei2，3 [1. China Resources Sanjiu(Ya’an) Pharmaceutical Co.， Ltd.， Sichuan Ya’an 625000， China; 2. Pharmacy College， Chengdu University of TCM， Chengdu 611137， China; 3. Institute of Innovative Medicine Ingredients of Southwest Specialty Medicinal Materials， Chengdu 611137， China]
ABSTRACT OBJECTIVE： To establish an HPLC fingerprint of Ginseng Radix et Rhizoma Rubra， and to optimize its processing technology. METHODS： HPLC method was adopted. The determination was performed on Waters SymmetryShieldTM RP18 column with mobile phase consisted of acetonitrile-water (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was set at 30 ℃， and the detection wavelength was 203 nm. The sample size was 10 μL. Using ginsenoside Rb1 as reference peak， HPLC fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra was established. The similarity of them was evaluated by using Similarity Evaluation System of TCM Chromatogram Fingerprint (2012 A edition) to confirm common peak. With steaming temperature， time and drying method as factors， using the content of ginsenoside and fingerprint similarity as index， the processing technology was optimized with L16(43) orthogonal test design and verified. Cluster analysis was conducted with SPSS 19.0 statistical software of 10 batches of Ginseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample. RESULTS： There were a total of 13 common peaks in the fingerprints of 10 batches of Ginseng Radix et Rhizoma Rubra. The similarity was more than 0.920; 3 common peaks were identified， such as ginsenoside Rg1， ginsenoside Re， ginsenoside Rb1. The optimal processing technology included that steamed at 100 ℃ for 150 min， dried at 60 ℃. The results of validation test show that the contents of ginsenoside Rg1， Re and Rb1 were 0.26%-0.29%， 0.17%-0.20%， 0.47%-0.54%， and the similarity between 3 batches of Ginseng Radix et Rhizome Rubra optimal processed sample and the control fingerprints was more than 0.970. The results of cluster analysis showed that 10 batches of Gimseng Radix et Rhizoma Rubra and 3 batches of optimal processed sample could be clustered into two categories; HS3-HS10 could be clustered into one category， and 3 batches of optimal processed sample， HS1 and HS2 be clustered into one category. CONCLUSIONS： Established fingerprint can be used for the optimization of processing technology of Gimseng Radix et Rhizoma Rubra， and characterize the correlation between fluctuation of technology parameter and quality of medicinal material; the optimal processing technology is reasonable and feasible., 百拇医药(侯新莲 黄露 彭成 朱雅宁 蔡帮军 周勤梅)