A Simple HPLC Method for Chlorogenic Acid Quantification in Jiruge-6 Suppositories-Scilight

Trends in Immunotherapy

Article

A Simple HPLC Method for Chlorogenic Acid Quantification in Jiruge-6 Suppositories

Downloads

https://doi.org/10.54963/ti.v9i2.908
Yilina, Yiran Bao, Nomin‑Erdene Ulambayar, & Enkhjargal Dorjbal. (2025). A Simple HPLC Method for Chlorogenic Acid Quantification in Jiruge-6 Suppositories. Trends in Immunotherapy, 9(2), 143–156. https://doi.org/10.54963/ti.v9i2.908
Volume. 9, Issue. 2 ( 2025)
Copyright (c) 2025 Yilina, Yiran Bao, Nomin‑Erdene Ulambayar, Enkhjargal Dorjbal
Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Copyright

The authors shall retain the copyright of their work but allow the Publisher to publish, copy, distribute, and convey the work.

License

Trends in Immunotherapy (TI)  publishes accepted manuscripts under Creative Commons Attribution 4.0 International (CC BY 4.0). Authors who submit their papers for publication by TI agree to have the CC BY 4.0 license applied to their work, and that anyone is allowed to reuse the article or part of it free of charge for any purpose, including commercial use. As long as the author and original source are properly cited, anyone may copy, redistribute, reuse, and transform the content.

Authors

  • Yilina

    Department of Pharmaceutical chemistry and Pharmacognosy, School of Pharmacy, Mongolian National University of Medical Sciences, Ulaanbaatar 16081, Mongolia
  • Yiran Bao

    Shenji Biotechnology Co., Ltd., Hohhot 010010, China
  • Nomin‑Erdene Ulambayar

    Department of Acupuncture and Moxibustion, International School of Mongolian Medicine, Mongolian National University of Medical Sciences, Ulaanbaatar 13270, Mongolia
  • Enkhjargal Dorjbal

    Department of Pharmaceutical chemistry and Pharmacognosy, School of Pharmacy, Mongolian National University of Medical Sciences, Ulaanbaatar 16081, Mongolia

Received: 22 April 2025; Revised: 19 May 2025; Accepted: 22 May 2025; Published: 9 June 2025

Chlorogenic acid (CGA) is a key bioactive polyphenol in Traditional Chinese Medicine (TCM) formulations including Jiruge-6 Suppositories. This study aimed to develop and validate a simple and sensitive HPLC method for quantifying CGA in these products. The optimized method utilized a C18 column with a mobile phase of acetonitrile and 0.1% phosphoric acid (15:85, v/v) at 1.0 mL/min flow rate, with detection at a 327 nm. Ultrasonic extraction with 70% methanol enhanced sample preparation efficiency. The method demonstrated excellent linearity (r² > 0.999) across the range of 1.0–100.0 μg/mL, high precision (RSD < 2.0%), and good recovery (98.2–101.5%). Detection and quantification limits were 0.30 μg/mL and 0.95 μg/mL, respectively. Analysis of five commercial batches from the same manufacturer revealed CGA content of 0.42-0.57 mg/suppository with notable seasonal variation. This validated method provides a practical approach for routine quality control of Jiruge-6 Suppositories, supporting standardization of TCM products.

Keywords:

Chlorogenic Acid; Jiruge‑6 Suppositories; HPLC; Method Validation; Traditional Chinese Medicine

References

  1. Bounegru, A.V.; Apetrei, C. Caffeic and Chlorogenic Acid in Coffee and Methods for Their Detection. In Coffee in Health and Disease Prevention, 2nd ed.; Elsevier: Amsterdam, Netherlands, 2025; pp. 893–904.
  2. Jiang, H.; G, Y.; Li, J.; et al. Inhibitory Mechanism of Chlorogenic Acid on α-Glucosidase and Evaluation of Its Glucose Consumption in HepG2 Cells. J. Mol. Struct. 2025, 141607.
  3. Kusumiyati, K.; Munawar, A.A.; Andasuryani, A. Investigating the Possibility of Using Near Infrared Spectroscopy (NIRS) Method in Coffee Beans Caffeine and Chlorogenic Acid (CGA) Determination by Near Infrared Spectroscopy (NIRS). Int. J. Hortic. Sci. Technol. 2025, 797–810.
  4. Ertekin, Z.C.; Köroğlu, A.; Dinç, E. Three‐Dimensional Spectrochromatographic Determination of Chlorogenic Acid in Melampyrum stenophyllum Boiss. Extracts by Parallel Factor Analysis. Phytochem. Anal. 2025, 36, 279–288.
  5. Han, X.; Chen, J.; G, Q.; Zhang, J.; et al. Photodynamically Activated Chlorogenic Acid-Based Antimicrobial Packaging Films for Cherry Preservation. Food Chem. 2025, 479, 143857.
  6. Zhao, P.; Song, Z.; Li, Y.; et al. Rapid and Simple Fluorescent Detection of Chlorogenic Acid in Aidi Injection Using Aggregation-Induced Emission (AIE) Nanoclusters. J. Pharm. Biomed. Anal. 2025, 254, 116570.
  7. Chen, J.; Lei, Q.; Yang, W.; et al. A Green and Rapid High‐Performance Liquid Chromatography–Ultraviolet Method for Determination of Four Organic Acids in Different Parts of Dandelion with Blue Applicability Grade Index Assessment. J. Food Biochem. 2025, 4077536.
  8. Tian, W.; Gonzales, G.B.; Wang, H.; et al. Caffeic Acid and Chlorogenic Acid Mediate the ADPN-AMPK-PPARα Pathway to Improve Fatty Liver and Production Performance in Laying Hens. J. Anim. Sci. Biotechnol. 2025, 16, 49.
  9. Jiang, X.; Li, L.; Wang, C.; et al. Dynamic/Static Pressure-Induced Copolymerization and Property Changes of Lotus Seed Starch with Chlorogenic Acid. Food Chem. 2025, 464, 141723.
  10. Ye, Z.; Cheng, L.; Xuan, Y.; et al. Chlorogenic Acid Alleviates the Development of Severe Acute Pancreatitis by Inhibiting NLPR3 Inflammasome Activation via Nrf2/HO-1 Signaling. Int. Immunopharmacol. 2025, 151, 114335.
  11. Talukder, P.; Chanda, S.; Sinha, B. Boosting Biotic Stress Resistance in Solanum melongena L.: The Role of Exogenous Chlorogenic Acid in Enhancing Secondary Metabolite Production. Appl. Biochem. Biotechnol. 2025, 1–24.
  12. Yu, R.; Li, P. Removal of Bisphenol A at Full-Scale Wastewater Treatment Plants and Antagonistic Effects of Chlorogenic Acid on It. Int. J. Environ. Sci. Technol. 2025, 1–14.
  13. Baby, S.; Alyami, N.M.; Aljawdah, H.M.; et al. Chlorogenic Acid Functionalized Tin Oxide-Sodium Alginate Hybrid Nanomaterials Induce Oxidative Stress-Mediated Apoptosis in Breast Cancer MDA-MB-231 Cells. Indian J. Pharm. Educ. Res. 2025, 59, 757–766.
  14. Nie, W.; Zhong, W.; Qian, L.; et al. Oral Chitosan-Cyclodextrin “Shell-Core” Nanoparticles Co-Loaded Rhein and Chlorogenic Acid for Ulcerative Colitis Treatment. Int. J. Biol. Macromol. 2025, 288, 138493.
  15. Pyrzynska, K. Spent Coffee Grounds as a Source of Chlorogenic Acid. Molecules 2025, 30, 613.
  16. Ibrahim, U.K.; Suzihaque, M.U.H.; Zaki, N.A.M.; et al. Microencapsulation of Chlorogenic from Green Coffee Bean Using Spray Drying. In Proceedings of the Sustainable and Integrated Engineering International Conference: SIE2022, Kuala Lumpur, Malaysia, 12–13 December 2022.
  17. Wu, S.; Zhou, Q.; Gao, Z.; et al. Chlorogenic Acid Targets SIRT6 to Relieve UVB-Induced UV Damage. Arch. Dermatol. Res. 2025, 317, 600.
  18. Ran, X.; Feng, L.; Hongying, F.; et al. Determination of Four Compositions of Lonicerae japonicae flos Based on UPLC–MS/MS Method. Pharmacogn. Mag. 2025, 09731296241265167.
  19. Yao, Y.X.; Yao, C.H.; Zhang, C.Y.; et al. Chlorogenic Acid Ameliorates Acetaminophen-Induced Liver Injury Through AMPK/mTOR/ULK1-Mediated Autophagy Activation. Am. J. Chin. Med. 2025, 1–20.
  20. Qu, W.; Deng, X.; Li, Y.; Zhang, X.; et al. Improving the Bioavailability and Synergistic Antioxidant Efficacy of Co-Embedded Chlorogenic Acid and Kaempferol Through an Ultrasonic-Prepared Zein-Pectin Nanoemulsion System. SSRN Electron. J. 2025, 1–40.
  21. Yu, L.; Zhang, X.; Guo, C.; et al. Inhibitory Effect of Chlorogenic Acid and Vanillic Acid on Fluorescent Advanced Glycation End Products Formation in Low-Temperature-Processed Pork Meat. Food Biosci. 2024, 62, 105041.
  22. Li, F.; Hou, H.; Zhao, T.; et al. Revealing the Non-Enzymatic Covalent Interaction Between Neo−/Crypto-Chlorogenic Acid and Beta-Lactoglobulin Under Nonthermal Process and Potential Delivery Capability. Food Chem. 2025, 143325.
  23. Wang, Q.; Du, T.; Zhang, Z.; Zhang, Q.; Zhang, J.; Li, W.; Hu, H.Y. Target Fishing and Mechanistic Insights of the Natural Anticancer Drug Candidate Chlorogenic Acid. Acta Pharm. Sin. B 2024, 14, 4431–4442.
  24. Zhang, Q.; Xu, Y.; Li, J.; et al. Construction and Dynamic In Vitro Digestive Characteristics of Whey Protein/Chlorogenic Acid/High Methoxy Pectin Water-in-Oil-in-Water Emulsion. Int. J. Biol. Macromol. 2025, 309, 142956.
  25. Olthof, M.R.; Hollman, P.C.; Katan, M.B. Chlorogenic Acid and Caffeic Acid Are Absorbed in Humans. J. Nutr. 2001, 131, 66–71.
  26. Naveed, M.; Hejazi, V.; Abbas, M.; et al. Chlorogenic Acid (CGA): A Pharmacological Review and Call for Further Research. Biomed. Pharmacother. 2018, 97, 67–74.
  27. Reverter, M.; Bontemps, N.; Lecchini, D.; et al. Use of Plant Extracts in Fish Aquaculture as an Alternative to Chemotherapy: Current Status and Future Perspectives. Aquaculture 2014, 433, 50–61.
  28. Liu, W.; Duan, Y.; Hu, X.; et al. Dual Effects of Dietary Lactobacillus helveticus and Chlorogenic Acid on Growth Performance, Digestibility, Immune-Antioxidant Capacity and Resistance Against Heat Stress of Juvenile Common Carp. Aquaculture 2022, 549, 737788.