Skip to main content
SpringerLink
Log in

A validated high-performance thin-layer chromatography method for the quantification of chlorogenic acid in the hydroalcoholic extract of Gynura cusimbua leaves

  • Original Research Paper
  • Published:
JPC – Journal of Planar Chromatography – Modern TLC Aims and scope Submit manuscript

Abstract

A simple, reliable, and reproducible high-performance thin-layer chromatography (HPTLC) method was developed for the quantitative determination of chlorogenic acid in the hydroalcoholic extract of Gynura cusimbua (D. Don) S. Moore leaves (HAGC). HPTLC plates (10 cm × 10 cm) pre-coated with silica gel 60F254 as the stationary phase and a mobile phase consisting of ethyl acetate‒formic acid‒acetic acid‒water (100:11:11:2.6, V/V) were used for achieving good separation. Densitometric evaluation for fingerprinting and quantification was performed at a scanning speed of 100 mm/s and 6 × 0.45-nm micro-slit dimension at 366 nm in absorption mode with a deuterium lamp. The method was validated in terms of linearity, limits of detection and quantification, precision, accuracy, and robustness parameters. The RF of chlorogenic acid was 0.43 ± 0.01. The linear regression analysis data for the calibration plots of chlorogenic acid showed a good linearity relationship with r2 = 0.9996, in the concentration range of 50‒250 ng/spot. The limits of quantification and detection were 43.12 and 14.36 ng/band, respectively. The amount of chlorogenic acid in HAGC was found to be 3.37% w/w. The quantification and the method validation of chlorogenic acid have not yet been reported for G. cusimbua which can be employed for the appropriate standardization of the plant extract/formulation.

Graphical Abstract

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

%RSD :

Percent relative standard deviation

HAGC:

Hydroalcoholic extract of Gynura cusimbua (D. Don) S. Moore leaves

HPTLC:

High-performance thin-layer chromatography

ICH:

International council for harmonisation

LOD:

Limit of detection

LOQ:

Limit of quantification

SD:

Standard deviation

References

  1. Leishangthem S, Sharma LD (2014) Study of some important medicinal plants found in Imphal-East District, Manipur India. Int J Sci Res Publ 4(9):1–5

    Google Scholar 

  2. Lakshman CD (2016) Medicinal and aromatic plants of North-East India. Int J Dev Res 6(11):10104–10114

    Google Scholar 

  3. Tewari B, Medhabati M (2019) Anti-ulcer effect of aqueous extract of Gynura cusimbua (D. Don) S. Moore on swimming stress ulcer model in albino rats. J Evol Med Dent Sci 8(33):2602–2607. https://doi.org/10.1460/jemds/2019/567

    Article  CAS  Google Scholar 

  4. Pradheep K, Pandey A, Bhatt KC (2016) Wild edible plants used by Konyak tribe in Mon district of Nagaland: survey and inventorisation. Indian J Nat Prod Resour 7(1):74–81. https://doi.org/10.56042/ijnpr.v7i1.9963

    Article  Google Scholar 

  5. Ma Q, Wei R, Zhou B, Sang Z, Liu W, Cao Z (2019) Antiangiogenic phenylpropanoid glycosides from Gynura cusimbua. Nat Prod Res 33(4):457–463. https://doi.org/10.1080/14786419.2017.1389931

    Article  CAS  PubMed  Google Scholar 

  6. Singh TS, Roy SS, Kshetri P, Ansari MA, Sharma SK, Verma MR, Singh IM, Prakash N, Kandpal B (2021) Comparative study on phenolic, flavonoids and in vitro antioxidant activity of wild edible plants from Loktak Lake wetland ecosystem under North East Indian Himalayan Region. Nat Prod Res 35(24):6045–6048. https://doi.org/10.1080/14786419.2020.1817014

    Article  CAS  PubMed  Google Scholar 

  7. Ma Q, Wei R, Zhong G, Sang Z (2020) Neuroprotective flavonoids from the aerial parts of Gynura cusimbua. Chem Nat Compd 56(4):725–728. https://doi.org/10.1007/s10600-020-03131-x

    Article  CAS  Google Scholar 

  8. Meng X, Li J, Li M, Wang H, Ren B, Chen J, Li W (2021) Traditional uses, phytochemistry, pharmacology and toxicology of the genus Gynura (Compositae): a comprehensive review. J Ethnopharmacol 276:114145. https://doi.org/10.1016/j.jep.2021.114145

    Article  CAS  PubMed  Google Scholar 

  9. Rana VS, Blazquez MA (2007) Chemical constituents of Gynura cusimbua aerial parts. J Essent Oil Res 19(1):21–22. https://doi.org/10.1080/10412905.2007.9699219

    Article  CAS  Google Scholar 

  10. Cai L, Feng Y (2020) Effects of exogenous ABA on the physiological characteristics and chlorophyll fluorescence of Gynura cusimbua seedlings under drought stress. InIOP Conf Ser: Earth Environ Sci 615(1):012089. https://doi.org/10.1088/1755-1315/615/1/012089

    Article  Google Scholar 

  11. Li J, Qin Y, Yu X, Xiong Z, Zheng L, Sun Y, Shen J, Guo N, Tao L, Deng Z, Liu X (2019) In vitro simulated digestion and in vivo metabolism of chlorogenic acid dimer from Gynura procumbens (Lour.) Merr.: enhanced antioxidant activity and different metabolites of blood and urine. J Food Biochem 43:e12654. https://doi.org/10.1111/jfbc.12654

    Article  CAS  PubMed  Google Scholar 

  12. Kim HH, Ha SE, Vetrivel P, Bhosale PB, Kim SM, Kim GS (2021) Potential antioxidant and anti-inflammatory function of Gynura procumbens polyphenols ligand. Int J Mol Sci 22:8716. https://doi.org/10.3390/ijms22168716

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Wang L, Pan X, Jiang L, Chu Y, Gao S, Jiang X, Zhang Y, Chen Y, Luo S, Peng C (2022) The biological activity mechanism of chlorogenic acid and its applications in food industry: a review. Front Nutr 9:943911. https://doi.org/10.3389/fnut.2022.943911

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Kristiningrum N, Retnaningtyas Y, Pertiwi NP (2015) Validated TLC-densitometry method for determination of chlorogenic acid in coffee leaves extract. Res J Pharm Biol Chem Sci 6(5):1138–1143

    CAS  Google Scholar 

  15. Urakova IN, Pozharitskaya ON, Shikov AN, Kosman VM, Makarov VG (2008) Comparison of high performance TLC and HPLC for separation and quantification of chlorogenic acid in green coffee bean extracts. J Sep Sci 31(2):237–241. https://doi.org/10.1002/jssc.200700472

    Article  CAS  PubMed  Google Scholar 

  16. Skalicka-Woźniak K, Hajnos M, Głowniak K (2009) High-performance thin-layer chromatography combined with densitometry for quantitative analysis of chlorogenic acid in fruits of Peucedanum alsaticum L. J Planar Chromat-Mod TLC 22(4):297–300. https://doi.org/10.1556/JPC.22.2009.4.11

    Article  CAS  Google Scholar 

  17. Shahnawaz PDK, Dey A (2018) HPTLC Analysis of the antioxidant and possible antidiabetic chlorogenic acid in the in situ and in vitro populations of the low-calorie sweetener Stevia rebaudiana (Bert.) Bertoni. Anal Chem Lett 8(6):872–881. https://doi.org/10.1080/22297928.2018.1515657

    Article  CAS  Google Scholar 

  18. Darekar DP, Sheikh MV, Hate MS (2018) High performance thin layer chromatography (HPTLC) and high performance liquid chromatography (HPLC) for qualitative and quantitative analysis of chlorogenic acid in Calendula officinalis Linn. Int J Sci Res 9(10):764–767. https://doi.org/10.21275/SR201008211148

    Article  Google Scholar 

  19. Biswas S, Das A, Roy D, Roy T, Dewan D, Paul RK, Bhowmick M, Nag M (2022) Developed and validated method for the simultaneous estimation of ferulic acid and chlorogenic acid in four Indian Ananas comosus merrill cultivars through HPTLC. J Med Pharm Allied Sci 11(1):4250–4255. https://doi.org/10.55522/jmpas.V11I1.1639

    Article  Google Scholar 

  20. Creţu GE, Morlock G, Grosu AR, Nechifor AC (2013) A high-performance thin-layer chromatographic method for chlorogenic acid and hyperoside determination from berry extracts. Rom Biotechnol Lett 18(5):8657–8665

    Google Scholar 

  21. Singh AP, Singh DP, Srivastava S, Govindarajan R, Rawat AK (2007) A validated quantitative HPTLC method for analysis of biomarkers in Ficus carica L. J Planar Chromat-Mod TLC 20:437–441. https://doi.org/10.1556/JPC.20.2007.6.8

    Article  CAS  Google Scholar 

  22. Sharma A, Gill N, Kumar R (2022) Development and validation of thin layer chromatography-densitometric method for quantification of kaempferol and chlorogenic acid in methanolic extract of Dragea volubilis. Asian Pac J Health Sci 9(4):112–116. https://doi.org/10.21276/apjhs.2022.9.4.23

    Article  Google Scholar 

  23. Maleš Ž, Šarić D, Bojić M (2013) Quantitative determination of flavonoids and chlorogenic acid in the leaves of Arbutus unedo L. using thin layer chromatography. J Anal Methods Chem 22013:385473. https://doi.org/10.1155/2013/385473

    Article  CAS  Google Scholar 

  24. Orfali R, Perveen S, Aati HY, Alam P, Noman OM, Palacios J, Al-Kurbi BS, Al-Taweel AM, Khan A, Mehmood R, Khan SI (2021) High-performance thin-layer chromatography for rutin, chlorogenic acid, caffeic acid, ursolic acid, and stigmasterol analysis in Periploca aphylla extracts. Separations 8(4):44. https://doi.org/10.3390/separations8040044

    Article  CAS  Google Scholar 

  25. Pradhan SK, Gupta RC, Goel R, Preet R (2017) Simultaneous determination of chlorogenic and caffeic acid in Siegesbeckia orientalis L. (Xi Xian) by a validated high-performance thin-layer chromatographic method. J Planar Chromat-Mod TLC 30(6):516–520. https://doi.org/10.1556/1006.2017.30.6.9

    Article  CAS  Google Scholar 

  26. Mukherjee PK, Bahadur S, Chaudhary SK, Kar A, Mukherjee K (2015) Quality related safety issue-evidence-based validation of herbal medicine farm to pharma. In: Mukherjee PK (ed) evidence-based validation of herbal medicine. Elsevier, Amsterdam, pp 1–28. https://doi.org/10.1016/B978-0-12-800874-4.00001-5

    Chapter  Google Scholar 

  27. Mukherjee PK, Kar A, Biswas S, Chaudhary SK, Banerjee S (2022) Hyphenated analytical techniques for validation of herbal medicine. In: Mukherjee PK (ed) Evidence-based validation of herbal medicine, 2nd edn. Elsevier, Amsterdam, pp 811–827. https://doi.org/10.1016/B978-0-323-85542-6.00038-X

    Chapter  Google Scholar 

  28. ICH Harmonised Tripartite Guideline (2005) Validation of analytical procedures: text and methodology Q2 (R1). In: Proceedings of the International Conference on Harmonization (ICH ‘05), Geneva

  29. Chaudhary SK, Lalvenhimi S, Biswas S, Chanda J, Kar A, Bhardwaj PK, Sharma N, Mukherjee PK (2022) High-performance thin-layer chromatography (HPTLC) method development and validation for the quantification of catechin in the hydroalcoholic extract of Parkia roxburghii seed. J Planar Chromat-Mod TLC 35(2):161–167. https://doi.org/10.1007/s00764-022-00164-6

    Article  CAS  Google Scholar 

Download references

Acknowledgements

The authors are thankful to the Institute of Bioresources and Sustainable Development (IBSD), Department of Biotechnology (DBT), Government of India, Imphal, Manipur, India, for the necessary help and support. The manuscript represents IBSD publications number as IBSD/MS/2020/01/110.

Funding

The authors are thankful to Department of Biotechnology, Government of India, for financial assistance bearing Grant No. BT/PR45281/NER/95/1934/2022.

Author information

Authors and Affiliations

  1. Institute of Bioresources and Sustainable Development (IBSD), Department of Biotechnology, Government of India, Takyelpat, Imphal, 795001, Manipur, India

    Sushil K. Chaudhary, Amit Kar, Pardeep K. Bhardwaj, Nanaocha Sharma, Sarangthem Indira Devi & Pulok K. Mukherjee

Authors
  1. Sushil K. Chaudhary
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Amit Kar
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Pardeep K. Bhardwaj
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nanaocha Sharma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Sarangthem Indira Devi
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Pulok K. Mukherjee
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pardeep K. Bhardwaj.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest with the article.

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chaudhary, S.K., Kar, A., Bhardwaj, P.K. et al. A validated high-performance thin-layer chromatography method for the quantification of chlorogenic acid in the hydroalcoholic extract of Gynura cusimbua leaves. JPC-J Planar Chromat 36, 45–53 (2023). https://doi.org/10.1007/s00764-023-00230-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00764-023-00230-7

Keywords

Search

Navigation