452 Summary A high-performance thin-layer chromatographic (HPTLC) meth- od for the simultaneous quantitative determination of cafeic acid, vanillic acid, syringic acid and kaempferol in lowers and buds of three diferent Bauhinia species was developed for the irst time in the case of these species. Methanol was found to be the best for the highest possible recovery of target analytes. For achieving good sep- aration, a mobile phase of toluene–ethyl acetate–formic acid (5:4:1, v/v) was used. The densitometric determination was carried out at 350 nm in relection–absorption mode. The calibration curves were linear in the range of 100–700 ng per spot for cafeic acid, vanillic acid, syringic acid, and kaempferol. The methanolic frac- tions of Bauhinia variegata L. lowers (BVFM) showed the highest amount of cafeic acid (0.08%), B. variegata L. buds (BVBM) and Bauhinia purpurea L. lowers (BPFM) showed the highest amount of kaempferol (1.53%), Bauhinia acuminata L. lowers showed the highest amount of vanillic acid (0.40%), and B. acuminata L. buds showed the highest amount of syringic acid (0.08%). The proposed method is simple, precise, speciic, accurate, less time-consuming, and cost-efective. The statistical analysis of data obtained proves that the method is reproducible and selective and can be used for routine analysis of reported phenolic compounds in crude drug and extracts. The simultaneous quantiication of these phenolic com- pounds has not yet been reported in the case of these species which may be utilized for the proper standardization of the drug. 1 Introduction Genus Bauhinia (family: Caesalpiniaceae) consists of over 250 species of trees and shrubs [1]. They are distributed in vari- ous geographic locations in tropical climates. Plants in genus Bauhinia have characteristic butterly-shaped leaves. Certain Bauhinia species have a long history of traditional medicinal applications for the treatment of diferent kinds of ailments, A. Gupta, S. Verma, S. Khatoon, and A.K.S. Rawat, Pharmacognosy and Ethnopharmacology Division, CSIR – National Botanical Research Institute, Lucknow – 226001, India; and A. Gupta and H. Dwivedi, School of Pharmacy, Babu Banarsi Das University, Faizabad Road, Lucknow, India. E-mail: rawataks@redifmail.com particularly diabetes, infections as well as pain and inlamma- tion [2, 3]. Bauhinia purpurea L., Bauhinia variegata L., and Bauhinia acuminata L., known as Kanchanar in Hindi, are lowering plant species commonly found in Southeast Asia [4, 18, 26]. The aerial parts of the plant are reported to contain amino ac- ids, lavonoids, steroids, terpenoids, tannins, lactones, glyco- lipids, glycosyl steroids, quinines, phenyl fatty ester, pacharin, bauhiniastatins, and b-sitosterol. Certain lavanones, dihydrod- ibenzoxepins, and bibenzyls were reported from the root of this plant [5–9]. A wide range of chemical compounds including mixture of phytol fatty esters, lutein, isoquercitin, astragalin, etc. were also isolated [10]. Five lavonoids isolated from the diferent parts of Bauhinia have been identiied as quercetin, rutin, apigenin, and apigenin 7-O-glucoside [22]. The chemical constituents isolated so far from the plant are b-sitosterol, lupeol, kaempferol-3-glucoside, tannins [23], quercitrin, heptatriacontan-12,13-diol, and do- tetracont-15-en-9-ol [24]. Simultaneous determination of major lavonoids (apigenin, quercetin, rutin, luteolin, and quercitrin) in B. variegata has been performed earlier [25]. Diferent parts of these plants have been used in traditional medicine to cure a host of illness such as body pain, rheuma- tism, fever, dropsy, skin diseases, septicemia, diarrhea, tonic, astringent, dysentery, hemorrhoids, piles, laxative, anthelmint- ic, antileprotic, antigoitrogenic, obesity, stomatitis, antidote for snake-poisoning, dyspepsia, latulence, and as carminative [11, 19–21]. Several activities like antidiabetic, antibacterial, wound-healing, antioxidant, antimalarial, antimycobacterial, antifungal, and anticancerous activities have been reported [8, 12–15]. The decoction of its bark and leaves is given in biliousness, stone in bladder, venereal diseases, leprosy, and asthma [27, 28]. Bark is used for the treatment of leucorrhoea and shyphilis [29]. Phytochemical studies on Bauhinia racemosa Lam., B. pur- purea Linn., involving preliminary phytochemical studies, physicochemical studies, quantitative estimation of primary and secondary metabolites, thin-layer chromatographic (TLC) study, and high-performance liquid chromatographic (HPLC) High-Performance Thin-Layer Chromatographic Analysis for the Simultaneous Quantiication of Four Phenolics in Flowers and Flower Buds of Bauhinia purpurea L., Bauhinia variegata L., and Bauhinia acuminata L. Abhishek Gupta, Shikhar Verma, Sayyada Khatoon, Harinath Dwivedi, and Ajay K.S. Rawat* Key Words: Bauhinia species Caffeic acid Kaempferol Syringic acid Vanillic acid Journal of Planar Chromatography 28 (2015) 6, 452–457 DOI: 10.1556/1006.2015.28.6.6 0933-4173/$ 20.00 © Akadémiai Kiadó, Budapest