Quality Control

Research of chromatographic fingerprint and the determination of multiple constituents and the primary evaluation of the quality grade of Banlangen granules based on the reference drug*

  • QIAN Xiu-yu ,
  • NIE Li-xing ,
  • YUAN Wen-peng ,
  • HU Xin-hua ,
  • CHANG Yan ,
  • YU Jian-dong ,
  • WEI Feng ,
  • MA Shuang-cheng
Expand
  • 1. Chinese Pharmaceutical Association, Beijing, 100050, China;
    2. National Institutes for Food and Drug Control, Beijing 102629, China;
    3. Shenzhen People's Hospital (The Second Clinical Medical College, Jinan University; The First Affiliated Hospital, Southern University of Science and Technology), Shenzhen 518020, China;
    4. Chinese Pharmacopeia Commission, Beijing 100061, China

Received date: 2024-01-29

  Online published: 2025-01-07

Abstract

Objective: To establish the determination methods for chromatographic fingerprint and multiple constituents, and then evaluate the quality grade of 103 batches of Banlangen granules that were from 55 manufacturers based on the reference drug. Methods: Ultra-high-performance liquid chromatography (UPLC) analysis was used to establish the fingerprint of Banlangen granules and quantitate the contents of 7 components-uridine, adenine, vernine, (R,S)-goitrin, adenosine, syringin and clemastanin B. The test was performed on the Waters ACQUITY UPLC HSS T3(100 mm×2.1 mm,1.8 μm) column with gradient elution using methanol-water at a flow rate of 0.2 mL·min-1. The injection volume was 2 μL, and the column temperature was constant at 30 ℃. Then, the results were detected using adopting a multi-wavelength detection mode. Next, the characteristic peaks and their belonging were clarified by comparing them with the reference drug. The sample fingerprint similarity and determination of contents were finally calculated, and the quality grades of Banlangen granules in each item were evaluated by using the reference drug. Results: The fingerprint was established, and 10 main characteristic peaks were identified. The similarities of the sample’s fingerprints ranged from 0.541 to 0.993, of which 101 batches were over 0.75 that were reached the second level, and 81 batches were over 0.90 that were reached the first level. The linear ranges of 7 components had good linear relationships within their respective ranges. The average recoveries (n=9) were 97.0%-104.7% with RSDs all below 3%. The precision, stability, and repeatability of methods were all good with RSDs all below 3%. Unified the sample specification to 5 g per bag, the content range of Banlangen granules samples was between 0.189-10.347 mg per bag. As a result, the content of 75 batches of samples reached the second level, and the content of 59 batches of samples reached the first level. Conclusion: The established methods are simple, accurate and rapid, which can be used for the quality control and grade evaluation of Banlangen granules, as well as the research paradigm of other traditional Chinese medicine preparations.

Cite this article

QIAN Xiu-yu , NIE Li-xing , YUAN Wen-peng , HU Xin-hua , CHANG Yan , YU Jian-dong , WEI Feng , MA Shuang-cheng . Research of chromatographic fingerprint and the determination of multiple constituents and the primary evaluation of the quality grade of Banlangen granules based on the reference drug*[J]. Chinese Journal of Pharmaceutical Analysis, 2024 , 44(10) : 1795 -1806 . DOI: 10.16155/j.0254-1793.2024-0064

References

[1] 中华人民共和国药典2020年版. 一部[S]. 2020: 1110
ChP 2020.Vol Ⅰ[S]. 2020: 1110
[2] 丁成福. 板蓝根颗粒治疗流行性感冒作用研究[J]. 中国民族民间医药杂志, 2011, 20(15):47
DING CF. Study on the effect of Banlangen granule in treating influenza[J]. Chin J Ethnomed Ethnopharm, 2011, 20(15):47
[3] 孙建立. 板蓝根颗粒治疗急性上呼吸道感染的临床分析[J]. 世界最新医学信息文摘, 2018(97):120
SUN JL. Clinical analysis of Banlangen granules in the treatment of acute upper respiratory tract infections [J]. World Latest Med Inf, 2018(97):120
[4] 张琼丽. 乳头瘤病毒感染患者应用外用壳聚糖抗菌膜联合内服板蓝根颗粒治疗的临床效果[J]. 中国医药科学, 2016, 6(1):94
ZHANG QL. Clinical effect on external used chitosan antibacterial film combined oral Isatis-root granule treatment on patients with papillomavirus infection[J]. China Med Pharm, 2016, 6(1):94
[5] 李金鸾. 磷酸奥司他韦颗粒联合板蓝根颗粒治疗小儿急性病毒性腮腺炎的效果分析[J]. 临床医学工程, 2018, 25(5):629
LI JL. Analysis on the effect of phosphate oseltamivir granules combined with Banlangen granules in the treatment of children with acute viral parotitis[J]. Clin Med Eng, 2018, 25(5):629
[6] 黄运柒. 磷酸奥司他韦联合板蓝根颗粒治疗甲型H1N1流感疗效观察与护理探讨[J]. 海峡药学, 2019, 31(1):233
HUANG YQ. Observation on the therapeutic effect and nursing exploration of the combination of oseltamivir phosphate and Banlangen granules in the treatment of influenza A (H1N1)[J]. Strait Pharm J, 2019, 31(1):233
[7] CHEN J, ZHU ZP, GAO TH, et al. Isatidis Radix and Isatidis Folium: a systematic review on ethnopharmacology phytochemistry and pharmacology[J]. J Ethnopharmacol, 2022, 283: 114648
[8] XIAO P, HUANG H, CHEN J, et al. In vitro antioxidant and anti-inflammatory activities of Radix Isatidis extract and bioaccessibility of six bioactive components after simulated gastro-intestinal digestion[J]. J Ethnopharmacol, 2014, 157: 55
[9] WANG X, XIE Y, HU X, et al. Qualitative and quantitative analysis of glucosinolates and nucleosides in Radix Isatidis by HPLC and liquid chromatography tandem mass spectrometry [J]. Acta Pharm Sin B, 2013, 3(5):337
[10] 邹华彬. 基于指纹图谱的生物体系中药内禀品质等级的数理理论判别[J]. 世界中医药, 2016, 11(9):1876
ZOU HB. Fingerprint spectra-based mathematical theory in determining the intrinsic quality grade of biological system of Chinese medicine [J]. World Chin Med, 2016, 11(9):1876
[11] TAO YI, GU XH, LI WD, et al. Techniques for biological fingerprinting of traditional Chinese medicine[J]. TrAC Tre Anal Chem, 2017, 97: 272
[12] LIU X, JIANG W, SU M, et al. Quality evaluation of traditional Chinese medicines based on fingerprinting[J]. J Sep Sci, 2020, 43(1):6
[13] CHEN J, GAO JY, SUN GX. Quantitative analysis combined with chromatographic fingerprint and antioxidant activities for the comprehensive evaluation of compound Danshen tablets[J]. J Sep Sci, 2017, 40(6):1244
[14] 聂黎行, 吴炎培, 刘静, 等. 中成药质量标准研究有关问题思考[J]. 药学学报, 2023, 58(8):2260
NIE LX, WU YP, LIU J, et al. Considerations on investigation on quality standard of Chinese patent medicine[J]. Acta Pharm Sin, 2023, 58(8):2260
[15] 聂黎行, 查祎凡, 胡晓茹, 等. 基于对照制剂的牛黄清胃丸全处方鉴别研究和等级初评价[J]. 中草药, 2018, 49(22):95
NIE LX, ZHA YF, HU XR, et al. Whole-ingredient identification and primary grade evaluation of Niuhuang Qingwei pills based on reference drug[J]. Chin Tradit Herb Drugs, 2018, 49(22):95
[16] 陈馥, 周颖仪, 李华, 等. 基于对照制剂的沉香化气丸多组分含量测定研究[J]. 药物分析杂志, 2019, 39(10):1771
CHEN F, ZHOU YY, LI H, et al. Study on multi-component determination of Chenxiang Huaqi pills based on reference drug[J]. Chin J Pharm Anal, 2019, 39(10):1771
[17] 林敬开, 聂黎行, 姚力, 等. 基于对照制剂的复方丹参片质量评价新模式探讨[J]. 药物分析杂志, 2019, 39(10):1751
LIN JK, NIE LX, YAO L, et al. Dscussion on a novel quality evaluation model for compound Danshen tablets based on reference drug[J]. Chin J Pharm Anal, 2019, 39(10):1751
[18] 邬秋萍, 许妍, 罗跃华, 等. 基于对照制剂的抗宫炎片质量评价新模式的探讨[J]. 药物分析杂志, 2019, 39(10):1762
WU QP, XU Y, LUO YH, et al. Discussion on a novel model for quality evaluation of Kanggongyan tablets based on reference drug[J]. Chin J Pharm Anal, 2019, 39(10):1762
[19] 聂黎行, 查祎凡, 何风艳, 等. 牛黄清胃丸对照制剂的建立[J]. 药物分析杂志, 2019, 39(10):1738
NIE LX, ZHA YF, HE FY, et al. Establishment of Niuhuang Qingwei pills reference drug[J]. Chin J Pharm Anal, 2019, 39(10):1738
[20] 聂黎行, 查祎凡, 左甜甜, 等. 基于ICP-MS和对照制剂的牛黄清胃丸中重金属及有害元素残留量测定及风险评估[J]. 中国中药杂志, 2019, 44(1):82
NIE LX, ZHA YF, ZUO TT, et al. Determination and risk assessment of heavy metals and harmful elements residues in Niuhuang Qingwei pills based on ICP-MS[J]. China J Chin Mater Med, 2019, 44(1):82
[21] 查祎凡, 聂黎行, 于健东, 等. 基于超高效液相色谱法和对照制剂的牛黄清胃丸指纹图谱研究和质量等级初评价[J]. 中国药学杂志, 2019, 54(17):1438
ZHA YF, NIE LX, YU JD, et al. Fingerprint study and primary quality evaluation of Niuhuang Qingwei pills based on UPLC and reference drug[J]. Chin Pharm J, 2019, 54(17):1438
[22] 聂黎行, 戴忠, 马双成. 中药对照制剂研制指导原则和技术要求[J]. 中国中药杂志, 2017, 42(19):3672
NIE LX, DAI Z, MA SC. Guideline principle and technical requirement for preparing traditional Chinese medicine reference drug[J]. China J Chin Mater Med, 2017, 42(19):3672
[23] 查祎凡, 聂黎行, 于健东, 等. 基于对照制剂和UPLC的牛黄清胃丸中黄柏的质量评价[J]. 食品与药品, 2020, 22(1):13
ZHA YF, NIE LX, YU JD, et al. Quality evaluation of Phellodendri Chinensis Cortex in Niuhuang Qingwei pills based on reference drug and UPLC[J]. Food Drug, 2020, 22(1):13
[24] 钟兰, 肖小武, 杨甲玺, 等. 基于对照制剂的复方鲜竹沥液指纹图谱和多组分含量测定研究[J]. 中国药学杂志, 2024, 58(2):178
ZHONG L, XIAO XW, YANG JX, et al. Fingerprint and multi-component determination of Fufang Xianzhuli Ye based on reference drugs[J]. Chin Pharm J, 2024, 58(2):178
[25] 吴良发, 袁铭铭, 徐菲, 等. 基于对照制剂的大七厘丸含量考察和等级初评价[J]. 中国药师, 2022, 25(4):113
WU LF, YUAN MM, XU F, et al. Content determination and primary grade evaluation of Daqili pills based on reference preparation[J]. China Pharm, 2022, 25(4):113
[26] 于新兰, 王雪, 严丽, 等. 基于对照制剂的灵芝制剂HPLC三萜指纹图谱化学计量学分析[J]. 中国药学杂志, 2020, 55(4):298
YU XL, WANG X, YAN L, et al. Chemometric analysis of HPLC fingerprint of triterpenoids in different Ganoderma preparations based on reference drug[J]. Chin Pharm J, 2020, 55(4):298
[27] 唐慧英, 鄢丹, 张少锋, 等. 基于凝集活性检测的板蓝根颗粒质量生物测定方法研究[J]. 药学学报, 2010, 45(4):479
TANG HY, YAN D, ZHANG SF, et al. Agglutinated activity bioassay method for the determination of antivirus potency of Banlangen granules[J]. Acta Pharm Sin, 2010, 45(4):479
[28] 叶军, 明安萍. 板蓝根颗粒药物血清对HepG2.2.15细胞分泌HBsAg、HBeAg的影响[J]. 湖北中医药大学学报, 2012, 14(6):10
YE J, MING AP. Effects of serum BanLanGen grain on secreting HBsAg and HBeAg of HepG2.2.15 cells[J]. J Hubei Univ Chin Med, 2012, 14(6):10
[29] 令红艳. 不同板蓝根制剂腺苷含量测定及其抗炎作用比较[J]. 中国实验方剂学杂志, 2012, 18(11):143
LING HY. Determination of different Isatidis Radix and its preparations adenosine correlation with anti-inflammatory effect[J]. Chin J Exp Tradit Med Form, 2012, 18(11):143
[30] 何洁英, 何洁宝, 王汝上. 板蓝根总生物碱HPLC指纹图谱[J]. 中国实验方剂学杂志, 2015, 21(12):62
HE JY, HE JB, WANG RS. Fingerprint of total alkaloids from Isatidis Radix by HPLC[J]. Chin J Exp Tradit Med Form, 2015, 21(12):62
[31] 王建敏, 李伟. 板蓝根颗粒中有效成分的测定及药理作用研究进展[J]. 中国医药导报, 2019, 16(18):49
WANG JM, LI W. Determination of effective components in Banlangen granules and research progress of pharmacological effects[J]. Chin Med Her, 2019, 16(18):49
[32] 钱秀玉, 聂黎行, 戴忠, 等. UPLC-MS/ELSD法快速检测板蓝根颗粒及组分中的糖类成分[J]. 药物评价研究, 2020, 43(7):1267
QIAN XY, NIE LX, DAI Z, et al. Rapid determination of sugars in Banlangen granule and its fractions by UPLC-MS/ELSD[J]. Drug Eval Res, 2020, 43(7):1267
[33] QIAN XY, NIE LX, DAI Z, et al. Determination of free amino acids in Banlangen granule and its fractions by solid phase extraction combined with ion-pair high performance liquid chromatography using a corona-charged aerosol detector(SPE-HPLC-CAD)[J]. Curr Pharm Anal, 2021, 7(17):838
[34] SHI Y, ZZHENG C, LI J, et al. Separation and quantification of four main chiral glucosinolates in Radix Isatidis and its granules using high-performance liquid chromatography/diode array detector coupled with circular dichroism detection[J]. Molecules, 2018, 23(6):1305
[35] 赵亚丽. 板蓝根药材中3种核苷高效液相法测定含量分析[J]. 实用中医药杂志, 2017, 33(12):1458
ZHAO YL. Determination of three kinds of nucleosides in Radix Isatidis by HPLC[J]. J Pract Trad Chin Med, 2017, 33(12):1458
[36] XIAO P, LI X, CHEN J, et al. Simultaneous quantification of three chemical types bioactive compounds in Radix Isatidis and its relevant pharmaceutical dosage forms by HPLC-DAD[J]. J Brazil Chem Soc, 2017, 28(7):1237
[37] CHEN M, GAN L, LIN S, et al. Alkaloids from the root of Isatis indigotica[J]. J Nat Prod, 2012, 75(6):1167
[38] 董福越, 徐科一, 黄远. HPLC同时测定板蓝根颗粒中2种关键抗病毒成分的含量[J]. 中国现代应用药学, 2019, 36(18):2245
DONG FY, XU KY, HUANG Y. Simultaneous determination of two antiviral components in Banlangen granules by HPLC[J]. Chin J Mod Appl Pharm, 2019, 36(18):2245
[39] NIE LX, WU YL, DAI Z, et al. Antiviral activity of Isatidis Radix derived glucosinolate isomers and their breakdown products against influenza A in vitro/ovo and mechanism of action[J]. J Ethnopharmacol, 2020, 251: 112550
[40] YANG Z, WANG Y, ZHENG Z, et al. Antiviral activity of Isatis indigotica root-derived clemastanin B against human and avian influenza A and B viruses in vitro[J]. Int J Mol Med, 2013, 31: 867
[41] QIAN XY, NIE LX, ZHAO H, et al. Discovery and molecular elucidation of the anti-influenza material basis of Banlangen granules based on biological activities and ultra-high performance liquid chromatography coupled with quadrupole-orbitrap mass spectrometry[J]. J Ethnopharmacol, 2022, 298: 115683
Outlines

/