Vol.:(0123456789) 1 3 JPC – Journal of Planar Chromatography – Modern TLC (2020) 33:457–462 https://doi.org/10.1007/s00764-020-00063-8 ORIGINAL RESEARCH PAPER Validated simultaneous HPTLC analysis of scopoletin and gallic acid in the methanolic fraction of Jatropha glandulifera Jyotsana Dwivedi 1,3  · Abhishek Gupta 2  · Sarvesh Paliwal 3  · A. K. S. Rawat 1 Received: 19 June 2020 / Accepted: 22 October 2020 / Published online: 20 November 2020 © Akadémiai Kiadó, Budapest, Hungary 2020 Abstract A high-performance thin-layer chromatography method for the simultaneous quantitative determination and validation of scopoletin and gallic acid in the methanolic fraction of Jatropha glandulifera L. was developed for the frst time. For achieving good separation, the mobile phase of toluene‒ethyl acetate‒glacial acetic acid (7.5:2.5:0.1, V/V) was used. Densitometric determination was carried out at 254 nm for scopoletin and gallic acid, in refection/absorption mode; the calibration curves were linear in the range of 100‒600 ng per spot. During the analysis, the methanolic fraction of J. glandulifera L. showed the presence of scopoletin (0.08%) and gallic acid (0.04%). The proposed method is simple, precise, specifc, and accurate. The obtained data can be used for the routine analysis of reported biomarkers in crude drug and extracts. The simultaneous quantifcation and method validation of these biomarkers have not yet been reported in J. glandulifera L. which may be utilized for the proper standardization of the plant. Keywords Gallic acid · HPTLC · Jatropha glandulifera · Scopoletin 1 Introduction Jatropha species are used in traditional folklore medicine to cure various ailments in Africa, Asia, and Latin America [1]. Their usage as traditional health remedies is popular in Asia, Latin America, and Africa [2]. The highly colored oil extracts of the trunk, branch, and roots of the shrub Jatropha glandulifera (Roxb.) have been used for many years in Pakistan as a balsam for the relief of rheumatic pains. The two pigments responsible for the dark-red color of the wood were isolated and identifed by a combination of spectroscopic and degradative techniques when the major pigment was found to be 3,3-dimethylacry- lylshikonin and the minor pigment acetyl shikonin [3]. Several known species from genus Jatropha have been reported for their medicinal uses and biological activities such as J. glandulifera, J. gossypiifolia, J. integerrima, and J. curcas. Casbene-type diterpenes are found in the roots of J. glandulifera [4]. The acetone extract of the fresh leaves and the chloroform extract of the dry leaves of J. glandu- lifera exhibited the best molluscicidal activity [5]. J. glan- dulifera seed oil and roots are used for sinuses, ringworm, antiparalytic, rheumatic pain, and as purgative. The leaves are used in asthma, bronchitis, as analgesic, emmenagogue, and scorpion-sting [6, 7]. J. glandulifera contains 27% oil having antimicrobial potential [8]. The qualitative phyto- chemical analysis showed primary and secondary metabo- lites like alkaloids, carbohydrates, fats, favonoids, lignin, lipids, polyphenols, proteins, starch, saponins, and tannins [9]. The leaf extracts, stem extract, roots extract, latex, and oil of J. curcas, J. glandulifera showed inhibitory action against plant pathogenic bacteria on plant pathogenic fungi and plant pathogenic bacteria [10, 11]. The present study deals with the identifcation and quantifcation of the secondary metabolites of J. glandulifera by high-performance thin- layer chromatography (HPTLC) analysis as there is a lack of phytochemical research on this plant. A simple, sensitive, and accurate HPTLC method has been developed for the estimation of scopoletin, and gallic acid in the methanolic extract of aerial parts of J. glandulifera. The simultaneous * A. K. S. Rawat pharmacognosynbri@gmail.com 1 Pharmacognosy and Ethnopharmacology Division, CSIR- National Botanical Research Institute, Lucknow, India 2 Hygia Institute of Pharmaceutical Education and Research, Lucknow, India 3 Department of Pharmacy, Banasthali Vidyapeeth, Banasthali, India