Characteristic Chromatogram and Index Component Contents of Substance Benchmark of Classic Formula Qingwei Power
-
摘要:
目的 建立经典名方清胃散物质基准的HPLC特征图谱以及含量测定方法, 控制其质量以保证后续制备。 方法 遵循古籍并结合前期制备工艺的考察, 制备15批清胃散物质基准, 采用HPLC建立物质基准特征图谱, 使用中药色谱指纹图谱相似度评价系统(2012版)计算相似度; 将5种药材中的7种指标性成分作为清胃散物质基准含量测定指标, 建立其HPLC含量测定方法, 并对15批样品进行含量测定。 结果 特征图谱相似度均≥0.975, 标定了13个共有峰, 对7个共有峰进行了化学成分指认, 分别为梓醇(1号峰)、阿魏酸(5号峰)、异阿魏酸(6号峰)、盐酸小檗碱(7号峰)、盐酸巴马汀(8号峰)、丹皮酚(12号峰)、藁本内酯(13号峰)。建立了2套指标性成分含量测定方法, 其一测定了梓醇、阿魏酸、异阿魏酸、丹皮酚的含量, 规定了4种成分含量应分别在1.48~2.76、0.14~0.26、0.48~0.90、1.72~3.19 mg·g-1之间; 其二测定了盐酸黄连碱、盐酸巴马汀、盐酸小檗碱的含量, 规定了3种成分含量应分别在0.65~1.21、0.57~1.07、1.86~3.45 mg·g-1之间。 结论 通过清胃散物质基准特征图谱及含量测定方法建立了清胃散物质基准的质量控制方法, 该方法简单可靠, 为后续质量控制及制剂开发提供依据及基础。 Abstract:OBJECTIVE To establish an HPLC characteristic spectrum and content determination method for the reference material of the classic formula Qingwei Powder, and control its quality to ensure subsequent preparation. METHODS Following ancient books and combining with the investigation of the previous preparation process, 15 batches of Qingwei Powder material standards were prepared. HPLC was used to establish the material standard feature map, and the similarity was calculated using the Chinese medicine chromatographic fingerprint similarity evaluation system (2012 version); seven indicator components from five medicinal herbs were used as the reference content determination indicators for Qingwei Powder, and an HPLC content determination method was established for 15 batches of samples. RESULTS The similarity of the characteristic spectra was≥0.975; 13 common peaks were calibrated; and 7 common peaks were identified for chemical composition, including catalpol (peak 1), ferulic acid (peak 5), isoferulic acid (peak 6), berberine hydrochloride (peak 7), palmatine hydrochloride (peak 8), paeonol (peak 12), and ligustilide (peak 13). Two sets of indicator component content determination methods have been established. Firstly, the contents of catalpol, ferulic acid, isoferulic acid, and paeonol were determined, and the four component contents were specified to be 1.48-2.76、0.14-0.26、0.48-0.90、1.72-3.19 mg·g-1, respectively; secondly, the contents of coptisine hydrochloride, palmatine hydrochloride, and berberine hydrochloride was determined, and it was specified that the content of the three components should be 0.65-1.21、0.57-1.07、1.86-3.45 mg·g-1, respectively. CONCLUSION A quality control method for the reference material of Qingwei Powder has been established through the characteristic spectrum and content determination method. This method is simple and reliable, providing a basis and foundation for subsequent quality control and formulation development. -
图 1 15批清胃散物质基准的HPLC特征图谱及对照特征图谱(R)
注: 1.梓醇; 5.阿魏酸; 6.异阿魏酸; 7.盐酸小檗碱; 8.盐酸巴马汀; 12.丹皮酚；13.藁本内酯
Figure 1. HPLC specific chromatogram of 15 batches of Qingwei Power substance benchmark and reference specific chromatogram (R)
图 2 7个特征峰在清胃散物质基准中的指认
注: 1.梓醇; 5.阿魏酸; 6.异阿魏酸; 7.盐酸小檗碱; 8.盐酸巴马汀; 12.丹皮酚；13.藁本内酯
Figure 2. Identification of seven common peaks in Qingwei Power substance benchmark
图 3 清胃散及各单味药物质基准特征图谱
注: 1.梓醇; 5.阿魏酸; 6.异阿魏酸; 7.盐酸小檗碱; 8.盐酸巴马汀; 12.丹皮酚；13.藁本内酯
Figure 3. HPLC specific chromatogram of Qingwei Power substance benchmark and single herb medicine
图 4 清胃散及阴性物质基准特征图谱
注: 1.梓醇; 5.阿魏酸; 6.异阿魏酸; 7.盐酸小檗碱; 8.盐酸巴马汀; 12.丹皮酚；13.藁本内酯
Figure 4. HPLC specific chromatogram of Qingwei Power substance benchmark and negative substance benchmark
表 1 清胃散物质基准中饮片产地信息
Table 1. Information on the origin of decoction pieces in the substance benchmark of Qingwei Powder
品名 批次 产地 品名 批次 产地 当归 20210413001 甘肃省定西市岷县 牡丹皮 2019040603 安徽亳州十九里镇 20210413002 甘肃省定西市岷县 2019040605 安徽亳州十九里镇 20210413003 甘肃省陇南市宕昌县 2019052601 安徽铜陵顺安镇凤凰山村 20210413004 甘肃省甘南藏族自治州临潭县 2019062901 陕西商洛商州区夜村镇 20210413005 甘肃省定西市岷县 2019070101 山西运城 黄连 HL-18002 重庆市石柱县湖镇三合村 升麻 2019051101 吉林省通化市柳河县 HL-18005 重庆市石柱县黄水镇万胜村 2019071501 辽宁省抚顺市新宾县 HL-18006 湖北省利川市忠路镇钟灵村 2019071503 辽宁省抚顺市新宾县 HL-18012 四川资阳乐至县全胜乡大石岩村 2019071507 黑龙江省牡丹江市绥阳林业局 HL-18015 四川乐山市金口河区永胜镇栀杆村 2019071510 黑龙江省牡丹江市绥阳林业局 生地黄(酒制) DH-19001 河南省武陟县西陶镇 DH-19003 河南省武陟县西陶镇 DH-19007 河南省温县南张羌镇 DH-19009 河南省温县南张羌镇 DH-19012 河南省温县南张羌镇 
下载: 导出CSV
表 2 15批清胃散物质基准饮片组合信息
Table 2. Information on the combination of 15 batches of Qingwei Powder substance benchmark decoction pieces
批次 当归 黄连 牡丹皮 升麻 地黄 S1 20210413003 HL-18015 2019070101 2019071503 DH-19003 S2 20210413001 HL-18002 2019040605 2019051101 DH-19009 S3 20210413004 HL-18006 2019062901 2019071501 DH-19001 S4 20210413005 HL-18012 2019052601 2019071507 DH-19012 S5 20210413002 HL-18005 2019040603 2019071510 DH-19007 S6 20210413003 HL-18015 2019070101 2019071503 DH-19001 S7 20210413001 HL-18002 2019040605 2019051101 DH-19003 S8 20210413004 HL-18006 2019062901 2019071501 DH-19007 S9 20210413005 HL-18012 2019052601 2019071507 DH-19009 S10 20210413002 HL-18005 2019040603 2019071510 DH-19012 S11 20210413001 HL-18002 2019040603 2019051101 DH-19001 S12 20210413002 HL-18005 2019040605 2019071501 DH-19003 S13 20210413005 HL-18006 2019070101 2019071503 DH-19007 S14 20210413003 HL-18012 2019062901 2019071507 DH-19009 S15 20210413004 HL-18015 2019052601 2019071510 DH-19012
下载: 导出CSV
表 3 15批清胃散物质基准相似度结果
Table 3. Similarities of 15 batches of Qingwei Powder substance benchmark
批次 相似度 批次 相似度 S1 0.986 S9 0.995 S2 0.978 S10 0.999 S3 0.996 S11 0.989 S4 0.995 S12 0.994 S5 0.999 S13 0.998 S6 0.995 S14 0.990 S7 0.975 S15 0.992 S8 0.994
下载: 导出CSV
表 4 清胃散物质基准中指标性成分含量测定方法学考察
Table 4. Methodological study on content determination of index components in Qingwei Power substance benchmark
成分 回归方程 R2 线性范围/(μg·mL-1) 平均加样回收率/% 梓醇 Y=2 508.8X-14 196 0.999 1 5.642~180.6 104.6 阿魏酸 Y=97 304X-7 706.5 0.999 9 1.423~45.54 99.31 异阿魏酸 Y=96 233X-7 933.5 0.999 9 2.669~85.40 102.5 丹皮酚 Y=30 872X-13 914 0.999 9 5.682~181.8 99.99
下载: 导出CSV
表 5 清胃散物质基准中指标性成分含量测定结果(mg·g-1)
Table 5. Content determination results of index components in Qingwei Powder substance benchmark (mg·g-1)
批次 梓醇 阿魏酸 异阿魏酸 丹皮酚 S1 2.20 0.24 0.89 2.65 S2 1.79 0.19 0.51 2.24 S3 2.71 0.22 0.87 2.94 S4 1.87 0.21 0.59 2.83 S5 1.95 0.19 0.66 2.60 S6 2.08 0.24 0.83 2.74 S7 1.76 0.18 0.47 2.09 S8 2.30 0.19 0.75 1.99 S9 1.98 0.20 0.56 2.78 S10 2.06 0.20 0.66 2.58 S11 2.24 0.19 0.72 1.65 S12 2.76 0.20 0.89 2.38 S13 1.63 0.17 0.67 2.01 S14 2.08 0.19 0.47 2.66 S15 2.30 0.22 0.79 2.62 x±s 2.12±0.32 0.20±0.02 0.69±0.15 2.45±0.37
下载: 导出CSV
表 6 清胃散物质基准中生物碱类指标性成分含量测定方法学考察
Table 6. Methodological study on content determination of alkaloid index components in Qingwei Powder substance benchmark
成分 回归方程 R2 线性范围/(μg·mL-1) 平均加样回收率/% 盐酸黄连碱 Y=37 768X+200.25 0.999 9 0.332 8~10.65 95.09 盐酸巴马汀 Y=46 427X+79.020 0.999 9 0.226 9~7.260 94.92 盐酸小檗碱 Y=46 869X+9919.5 0.999 9 1.263~40.42 105.1
下载: 导出CSV
表 7 清胃散物质基准中生物碱类指标性成分含量测定结果(mg·g-1)
Table 7. Content determination results of alkaloid index components in Qingwei Powder substance benchmark(mg·g-1)
批次 盐酸黄连碱 盐酸巴马汀 盐酸小檗碱 S1 0.71 0.60 2.01 S2 1.00 0.86 2.76 S3 0.97 0.81 2.70 S4 0.98 0.96 2.93 S5 1.11 1.02 3.18 S6 0.85 0.68 2.30 S7 0.97 0.81 2.66 S8 1.17 0.94 3.19 S9 0.89 0.89 2.71 S10 0.85 0.78 2.42 S11 1.06 0.83 2.85 S12 0.94 0.81 2.68 S13 0.98 0.85 2.77 S14 0.84 0.82 2.56 S15 0.68 0.58 1.97 x±s 0.93±0.13 0.82±0.12 2.65±0.36
下载: 导出CSV
-
[1] 国家药品监督管理局. 中药注册分类及申报资料要求政策解读[EB/OL]. (2020-09-30)[2023-12-28]. https://www.nmpa.gov.cn/xxgk/zhcjd/zhcjdyp/20200930164259184.html?type=pc&m= .National Medical Products Administration. Interpretation of policies on classification and application requirements for traditional Chinese medicine registration[EB/OL]. (2020-09-30)[2023-12-28].https://www.nmpa.gov.cn/xxgk/zhcjd/zhcjdyp/20200930164259184.html?type=pc&m= .[2] 陈畅, 程锦堂, 刘安. 经典名方研发策略[J]. 中国中药杂志, 2017, 42(9): 1814-1818.CHEN C, CHENG J T, LIU A. Research and development strategies in classical herbal formulae[J]. China J Chin Mater Med, 2017, 42(9): 1814-1818. [3] 覃艺. 经典名方桂枝芍药知母汤物质基准的研究[D]. 成都: 成都中医药大学, 2019.QIN Y. The study on substance benchmarks of classical famous prescriptions Guizhi-Shaoyao-zhimu Decoction[D]. Chengdu: Chengdu University of TCM, 2019. [4] 李东垣. 兰室秘藏[M]. 李仲平, 校注. 北京: 中国医药科技出版社, 2011: 43.LI D Y. Secrets from the orchid chamber[M]. LI Z P proofreaded. Beijing: China Medical Technology Press, 2011: 43. [5] 姚杭琦, 龚苏晓, 李丹, 等. 经典名方清胃散的研究进展[J]. 药物评价研究, 2019, 42(7): 1474-1479.YAO H Q, GONG S X, LI D, et al. Research progress on Qingwei San of classical herbal formulae[J]. Drug Eval Res, 2019, 42(7): 1474-1479. [6] 聂欣, 庞兰, 江华娟, 等. 经典名方化肝煎物质基准特征图谱及多指标成分含量测定研究[J]. 中草药, 2020, 51(20): 5177-5186. doi: 10.7501/j.issn.0253-2670.2020.20.009NIE X, PANG L, JIANG H J, et al. Study on specific chromatogram and content determination of multi-index components about material reference of classic prescription of Huaganjian[J]. Chin Tradit Herb Drugs, 2020, 51(20): 5177-5186. doi: 10.7501/j.issn.0253-2670.2020.20.009 [7] 江雪欣, 梁伟娜. HPLC法同时测定妇科千金片中6种活性成分[J]. 中国药师, 2016, 19(11): 2174-2176. doi: 10.3969/j.issn.1008-049X.2016.11.054JIANG X X, LIANG W N. Simultaneous determination of six active components in fuke Qianjin Tablets by HPLC[J]. China Pharm, 2016, 19(11): 2174-2176. doi: 10.3969/j.issn.1008-049X.2016.11.054 [8] 杨净尧. 经典名方保元汤的物质基准研究[D]. 长春: 长春中医药大学, 2020.YANG J Y. Study on the material benchmark of the classic famous prescriptions Baoyuan Decoction[D]. Changchun: Changchun University of Chinese Medicine, 2020. [9] 张欣舒, 董金香, 杨净尧, 等. 经典名方保元汤物质基准指纹图谱及多指标量值传递研究[J]. 药物分析杂志, 2021, 41(2): 345-358.ZHANG X S, DONG J X, YANG J Y, et al. HPLC fingerprint of substance benchmarks of classical famous prescriptions Baoyuan Decoction and multi-component value transmitting[J]. Chin J Pharm Anal, 2021, 41(2): 345-358. [10] 郑玥, 凌雪宇, 刘文亮. 黄芪桂枝五物颗粒中芍药苷的含量测定[J]. 生物技术世界, 2013, 10(12): 61.ZHENG Y, LING X Y, LIU W L. Determination of paeoniflorin in Huangqi Guizhiwuwu granules[J]. Biotech World, 2013, 10(12): 61. [11] 张琴, 王佳丽, 高喜梅, 等. 经典名方温胆汤水煎液HPLC指纹图谱与多成分含量测定研究[J]. 南京中医药大学学报, 2021, 37(6): 930-937. doi: 10.14148/j.issn.1672-0482.2021.0930ZHANG Q, WANG J L, GAO X M, et al. Study on HPLC fingerprint and multi-component content determination of classical prescriptions Wendan Decoction[J]. J Nanjing Univ Tradit Chin Med, 2021, 37(6): 930-937. doi: 10.14148/j.issn.1672-0482.2021.0930 -
点击查看大图
计量
- 文章访问数: 64
- HTML全文浏览量: 10
- PDF下载量: 10
- 被引次数: 0
