Scopus (CiteScore 2022 =3.0, Q3) , ISC

Document Type : Original Research Article


1 Department of Pharmacy, Faculty of Medical and Health Science, Universitas Islam Negeri Maulana Malik Ibrahim Malang, East Java, Indonesia

2 Department of Biomedical Sciences, Faculty of Medicine and Health Science, Universitas Islam Negeri Maulana Malik Ibrahim Malang, East Java, Indonesia

3 Indonesia National Agency of Drug and Food Control, Ambon Maluku, Indonesia



This study aimed to develop an HPTLC-Densitometry method for identifying and determining the 1,4-naphthoquinone compound in ethanol and water extracts of Eleutherine palmifolia (EP). The method was validated for linearity, accuracy, precision, Limit of Detection (LOD), and Limit of Quantification (LOQ). The linearity is assessed to ensure a linear relationship between the instrument response and various analytical concentrations. Accuracy is measured by comparing the measurement results with reference values or true values. Precision is evaluated through repeated measurements on samples with the same concentration, both within (intra-assay) and between assays (inter-assay). The Limit of Detection (LOD) determines the lowest concentration that can be detected, while the Limit of Quantification (LOQ) determines the lowest concentration that can be measured and quantified accurately with a specific level of confidence. The linearity test yielded an r-value of 0.9976, with an LOD value of 163.6006 ppm and a LOQ value of 495.7595 ppm. The method's accuracy ranged from 80-120%, and the precision value from %RSD was 0.99%. The 1,4-naphthoquinone content in EP ethanol extract was found to be 0.318% (w/w), while in water extract it was 0.092% (w/w). The method meets standard validation requirements and is recommended for use in traditional medicine development due to its high concentration of 1,4-naphthoquinone.

Graphical Abstract

Standardizing 1,4-naphthoquinone measurement in Eleutherine Palmifolia: An hptlc-densitometry approach


Main Subjects

[1] M. Marlin, M. Simarmata,  U. Salamah, W. Nurcholis, Effect of nitrogen and potassium application on growth, total phenolic, flavonoid contents, and antioxidant activity of Eleutherine palmifolia,  AIMS Agriculture & Food2022, 7, 580–593.  [Crossref], [Google Scholar], [Publisher]
[2] R. Mutiah, R.A. Sari,  W.Y. Firsyaradha,  A. Listiyana,  Y.Y.A. Indrawijaya,  A. Wafi, A. Suryadinata, R. Susilowati, A. Rahmawati, Activity and toxicity of Eleutherine palmifolia (L.) Merr. extract on balb/c mice colitis-associated colon cancer model,  Asian Pacific Journal of Cancer Prevention: APJCP, 2020,  21, 3579–3586. [Crossref], [Google Scholar], [Publisher]
[3] A. Biworo, L.W. Atanta, I.S. Arianto, S. Hamidah, E. Suhartono, June. Ameliorative effect of tuber extract from bawang dayak (Eleutherine palmifolia (L.) merr) against acute UV-induced skin oxidative damage in Rattus norvegicus. In AIP Conference Proceedings , AIP Publishing, 2019, 2108, 020010. [Crossref], [Google Scholar], [Publisher]
[4] T.D. Harlita, A. Asnani, The antibacterial activity of dayak onion (Eleutherine palmifolia (L.) merr) towards pathogenic bacteria, Tropical life sciences research2018, 29, 39-52. [Crossref], [Google Scholar], [Publisher]
[5] M. Insanu, S. Kusmardiyani, R. Hartati, Recent Studies on Phytochemicals and Pharmacological Effects of Eleutherine americana Merr,  Procedia Chemistry2014, 13, 221-228 [Crossref], [Google Scholar], [Publisher]
[6] B. Gunjal Sanket, P.R. Dighe, Analysis of herbal drugs by HPTLC: A review, Asian Journal of Pharmaceutical Research and Development, 2022, 10, 125-128. [Crossref], [Google Scholar], [Publisher]
[7] M. Kabiri,  H. Rezadoost, A. Ghassempour,  A comparative quality study of saffron constituents through HPLC and HPTLC methods followed by isolation of crocins and picrocrocin. LWT2017, 84, 1-9. [Crossref], [Google Scholar], [Publisher]
[8] R. Annisa, E. Hendradi, M. Yuwono, February. Analysis of 1, 4 naphthoquinone in the Indonesian medical plant from extract Eleutherine palmifolia (L.) Merr by UHPLC, In IOP Conference Series: Earth and Environmental Science Publishing, 2020, 456, 12020. [Crossref], [Google Scholar], [Publisher]
[9] A.A. Kamarudin, N.H. Sayuti, N. Saad, N.A.A. Razak, N.M. Esa, Eleutherine bulbosa (Mill.) Urb. Bulb: Review of the pharmacological activities and its prospects for application, International Journal of Molecular Sciences2021, 22, 6747. [Crossref], [Google Scholar], [Publisher]
[10] R. Mutiah, C.M. Hadya, W.S. Bhagawan, R. Annisa, Y.Y.A. Indrawijaya, F.I. Huwaida, R.R. DA, R. Susilowati, I. Taufik, Metabolite Profiling of Eleutherine palmifolia (L.) Merr. By HPTLC-Densitometry and its Correlation with Anticancer Activities and In Vitro Toxicity, Indonesian Journal of Pharmacy2019, 30, 157-166. [Crossref], [Google Scholar], [Publisher]
[11] S. Shukla, Freeze drying process: A review, International journal of pharmaceutical sciences and research, 2011, 2, 3061-3068. [Google Scholar], [Publisher]
[12] S.H. Nile, S.W. Park, HPTLC densitometry method for simultaneous determination of flavonoids in selected medicinal plants, Frontiers in Life Science, 2015, 8, 97-103. [Crossref], [Google Scholar], [Publisher]
[13] P. Swarnkar, M.K. Gupta, M. Maheshwari, Analytical method validation of compendial hplc method for pharmaceuticals as per recent usp and ich guidelines, 2021. [Google Scholar], [Publisher]
[14] I.H.T. Guideline, Validation of analytical procedures: text and methodology, 2005,   1, 5. [Google Scholar], [Publisher]
[15] A. Ahmad, M. Amir, A.A. Alshadidi, M.D. Hussain, A. Haq, M. Kazi, Central composite design expert-supported development and validation of HPTLC method: Relevance in quantitative evaluation of protopine in Fumaria indica, Saudi Pharmaceutical Journal2020, 28, 487-494. [Crossref], [Google Scholar], [Publisher]
[16] A. Gantait, S. Pandit, N.K. Nema, P.K. Mukjerjee, Quantification of glycyrrhizin in Glycyrrhiza glabra extract by validated HPTLC densitometry, Journal of AOAC International2010, 93, 492-495. [Crossref], [Google Scholar], [Publisher]
[17] D.A.  Frommenwiler, A. Booker, R. Vila, M. Heinrich, E. Reich, S. Cañigueral, Comprehensive HPTLC fingerprinting as a tool for a simplified analysis of purity of ginkgo products, Journal of Ethnopharmacology2019, 243, 112084. [Crossref], [Google Scholar], [Publisher]
 [18] B. Mehta, S. Morge, HPTLC-densitometric analysis of candesartan cilexetil and hydrochlorothiazide in tablets, JPC-Journal of Planar Chromatography-Modern TLC2008, 21, 173-176. [Crossref], [Google Scholar], [Publisher]
[19] M.B. Patel, K.M. Patel, G.S. Patel, B.N. Suhagia, A.M. Prajapati, Development and validation of a stability‐indicating HPTLC‐Densitometric method for satranidazole, Journal of liquid chromatography & related technologies, 2007, 30, 2755-2767. [Crossref], [Google Scholar], [Publisher]
[20] D.H. Shewiyo, E.A.K.K. Kaale, P.G.  Risha, B. Dejaegher, J. Smeyers-Verbeke, Y. Vander Heyden, HPTLC methods to assay active ingredients in pharmaceutical formulations: A review of the method development and validation steps, Journal of pharmaceutical and biomedical analysis2012, 66, 11-23. [Crossref], [Google Scholar], [Publisher]
[21] R. Singh, HPLC method development and validation-an overview, Journal of Pharmaceutical Education & Research2013, 4, 26. [Google Scholar], [Publisher]
[22] M.H. Alqarni, A.I. Foudah, A. Alam, M.A. Salkini, P. Alam, H.S. Yusufoglu, Novel HPTLC-densitometric method for concurrent quantification of linalool and thymol in essential oils, Arabian Journal of Chemistry2021, 14, 102916. [Crossref], [Google Scholar], [Publisher]