Objective  To establish the HPLC fingerprint and a multi-indicator method for the content determination of the components of the Guifu Dihuang pills (concentrated pill), and provide reference for quality evaluation and standard research of Guifu Dihuang pills.

Methods  The separation was developed on Welch Ultimate AQ-C18 column (250 mm×4.6 mm, 5 μm) at 30 ℃ by gradient elution, and the mobile phase was acetonitrile-0.1% phosphoric acid with a flow rate of 1.0  mL/min. The detection wavelengths were 240 nm during 0-45 min, and 205 nm during 45-85 min. The “Similarity Evaluation System of Chromatographic Fingerprints of Traditional Chinese Medicine” (Version 2012) was used to evaluate the similarity of the HPLC fingerprints of 17 batches of samples and determine their contents.

Results  HPLC fingerprint of 17 batches of Guifu Dihuang pills (concentrated pill) was established, and the similarities was all greater than 0.95. A total of 15 common peaks were indentified, and 6 main characteristic peaks were identified, including morroniside, loganin, paeoniflorin, cinnamaldehyde, paeonol, and alisol B 23-acetate. These six components had good linear relationships within a certain concentration range (r≥0.999 4), with average recovery rates of 97.7% (RSD=1.9%), 96.5% (RSD=2.7%), 97.6% (RSD=1.3%), 98.4% (RSD=2.6%), 97.2% (RSD=2.0%), and 96.2% (RSD=2.4%) (n=6), respectively. The contents of 6 components in 17 batches of samples were 0.559-1.349, 1.042-1.693, 0.836-1.424, 0.110-0.455, 1.608-2.639, 0.202-0.309 mg/g, respectively.

Conclusion   The established fingerprint and content determination method of Guifu Dihuang pills are simple, sensitive and reproducible, which can be used as the basis for the follow-up quality standard research of Guifu Dihuang pills (concentrated pill).

1.刘晓晴, 孙磊, 乔善义. 市售桂附地黄丸质量的高效液相指纹图谱法评价[J]. 时珍国医国药, 2015, 26(2): 304-306. [Liu XQ, Sun L, Qiao SY. Evaluation on quality of Guifu Dihuang Wan by HPLC fingerprint method[J]. Lishizhen Medicine and Materia Medica Research, 2015, 26(2): 304-306.] DOI: 10.3969/j.issn.1008-0805.2015.02.019.

2.曹富, 徐宁, 刘慧娟. GC-MS技术在中药谱效关系研究中的应用进展[J]. 云南化工, 2022, 49(12): 1-3. [Cao F, Xu N, Liu  HJ. Application progress of GC-MS in the study of profile-effect of traditional Chinese medicine[J]. Yunnan Chemical Technology, 2022, 49(12): 1-3.] DOI: 10.3969/j.issn.1004-275X.2022.12.01.

3.刘诗琪, 杨超茅, 苟小军. 糖肾地黄汤HPLC指纹图谱及多成分含量测定方法[J]. 中国医院药学杂志, 2024, 44(16): 1890-1896. [Liu SQ, Yang CM, Gou XJ. High performance liquid chromatographic fingerprinting of Tangshen Dihuang decoction and measurements of its multiple components[J]. Chinese Journal of Hospital Pharmacy, 2024, 44(16): 1890-1896.] DOI: 10.13286/j.1001-5213.2024.16.07.

4.石征蓉, 杨秀青, 谷江华, 等. 基于标准汤剂指纹图谱分析的糖肾清毒颗粒质量评价[J]. 中药材, 2018, 41(1): 129-135. [Shi  ZR, Yang XQ, Gu JH, et al. Quality evaluation of tangshen qingdu granular based on the fingerprint analysis of standard decocion[J]. Journal of Chinese Medicinal Materials, 2018, 41(1): 129-135.] DOI: 10.13863/j.issn1001-4454.2018.01.027.

5.陆丹, 池玉梅, 高秋芳, 等. 虎掌南星HPLC指纹图谱建立[J]. 中成药, 2020, 42(10): 2788-2791. [Lu D, Chi YM, Gao QF, et al. Establishment of HPLC fingerprints of pinellia pedatisecta[J]. Chinese Traditional Patent Medicine, 2020, 42(10): 2788-2791.] DOI: 10.3969/j.issn.1001-1528.2020.10.048.

6.朱红磊, 崔永霞, 陈随清. 基于HPLC指纹图谱及化学计量学揭示不同贮存年限山茱萸的异同[J]. 中药材, 2021, 44(7): 1614-1619. [Zhu HL, Cui YX, Chen SQ. The similarities and differences of Cornus officinalis in different storage years were revealed based on HPLC fingerprint and chemometrics[J]. Journal of Chinese Medicinal Materials, 2021, 44(7): 1614-1619.] DOI: 10.13863/j.issn1001-4454.2021.07.013.

7.刘诗琪, 吴闯, 朱智敏, 等. 加味增液汤HPLC指纹图谱及3种主要成分的含量测定[J]. 中国药师, 2024, 27(6): 928-936. [Liu SQ, Wu C, Zhu ZM, et al. HPLC fingerprint and three main components determination of Modified Zengye decoction[J]. China Pharmacist, 2024, 27(6): 928-936.] DOI: 10.12173/j.issn.1008-049X.202311200.

8.聂格, 李晓玲, 石雨荷, 等. 健脾扶正颗粒HPLC指纹图谱的建立及8个化学成分含量测定[J]. 湖南中医药大学学报, 2023, 43(9): 1609-1616. [Nie G, Li XL, Shi YH, et al. Establishment of HPLC fingerprints and content determination of eight chemical components of Jianpi Fuzheng granules[J]. Journal of Hunan University of Chinese Medicine, 2023, 43(9): 1609-1616.] DOI: 10.3969/j.issn.1674-070X.2023.09.009.

9.白艳秋, 阿伊萨, 牧其尔, 等. HPLC法同时测定蒙药清肝玛努-4醇提物中6种成分的含量及指纹图谱的建立[J]. 中国药师, 2024, 27(5): 739-746. [Bai YQ, A YS, Mu QE, et al. Simultaneous determination of six compounds in the alcohol extract from Mongolian medicine Qinggan Manu-4 and the establishment of fingerprints by HPLC[J]. China Pharmacist, 2024, 27(5): 739-746.] DOI: 10.12173/j.issn.1008-049X.202401199.

10.赵晓梅, 田双双, 刘珊珊, 等. 关于《中国药典》2020年版泽泻质量标准修订的建议[J]. 中国中药杂志, 2020, 45(7): 1566-1577. [Zhao XM, Tian SS, Liu SS, et al. Recommendations on quality standards of Alismatis rhizoma in Chinese Pharmacopoeia (2020 edition)[J]. China Journal of Chinese Materia Medica, 2020, 45(7): 1566-1577.] DOI: 10.19540/j.cnki.cjcmm.20200202.202.

11.王海玲, 于泓苓, 张大奎, 等. 四妙勇安汤的高效液相色谱指纹图谱及含量测定[J]. 化学分析计量, 2024, 33(6): 61-67. [Wang HL, Yu HL, Zhang DK, et al. Determination of high performance liquid chromatography fingerprint chromatograms and content of Simiao Yong'an decoction[J]. Chemical Analysis and Meterage, 2024, 33(6): 61-67.] DOI: 10.3969/j.issn.1008-6145. 2024.06.011.

12.李蕊, 陈文莉, 康点点, 等. 川西小黄菊HPLC指纹图谱建立及7种成分含量测得[J]. 中成药, 2024, 46(6): 1794-1799. [Li R, Chen WL, Kang DD, et al. Establishment of HPLC fingerprints and content determination of seven constituents for Tanacetum tatsienense[J]. Chinese Traditional Patent Medicine, 2024, 46(6): 1794-1799.] DOI: 10.3969/j.issn.1001-1528.2024.06.004.

13.范鑫玉, 吉田, 笔雪艳, 等. 中药指纹图谱分类及评价概述[J]. 黑龙江医药, 2023, 36(6): 1254-1258. [Fan XY, Ji T, Bi XY, et al. Overview of the classification and evaluation of fingerprints of traditional Chinese medicines[J]. Heilongjiang Medicine Journal, 2023, 36(6): 1254-1258.] DOI: 10.14035/j.cnki.hljyy.2023. 06.004.

14.郭守斌. 中药指纹图谱应用进展分析与研究[J]. 世界最新医学信息, 2016, 16(96): 31-32. [Guo SB. Analysis and research on the application of TCM fingerprint[J]. World Latest Medicne Information, 2016, 16(96): 31-32.] DOI: 10.3969/j.issn.1671-3141.2016.96.022.

15.孙国祥, 孙万阳, 闫慧, 等. 中药整体质量控制标准体系构建和中药一致性评价步骤[J]. 中南药学, 2019, 17(3): 321-331. [Sun GX, Sun WY, Yan H, et al. Constructing traditional Chinese medicine standard system for overall quality control and quality consistency evaluation of Chinese medicine[J]. Central South Pharmacy, 2019, 17(3): 321-331.] DOI: 10.7539/j.issn.1672-2981.2019.03.001.