Search Results

You are looking at 1 - 10 of 17 items for :

  • "Betulinic acid" x
  • All content x
Clear All

Separation of three triterpenic acids (ursolic, oleanolic and betulinic acid) was achieved on different thin-layer chromatography (TLC) (silica gel 60) and high-performance thin-layer chromatography (HPTLC) sorbents (silica gel 60, C2 RP and C18 RP) using several developing solvents, based on the non-polar diluent n-hexane, and ester (methyl acetate, ethyl acetate, ethyl propionate) as selector. Anisaldehyde and molybdophosphoric acid detection reagents were used. Finally, a simple method on a C18 RP HPTLC plate was developed using n-hexane-ethyl acetate (5:1 v/v) as a developing solvent in a horizontal developing chamber. The method was used for the screening of ursolic, oleanolic and betulinic acids in different vegetable extracts. Other plant triterpenoids (lupeol, α-amyrin, β-amyrin, cycloartenol, lupenone, friedelin, lupeol acetate, cycloartenol acetate) and phytosterols (β-sitosterol, stigmasterol) did not interfere. TLC-MS was used as a tool for the additional confirmation of the presence of ursolic, oleanolic, and betulinic acids in some of the studied vegetable extracts. Ursolic and oleanolic acids were found in radicchio Leonardo and white-colored radicchio di Castelfranco extracts for the first time, while betulinic acid was not detected in the eggplant extract by MS, although it was suggested at first by TLC analysis. Pre-chromatographic bromination on the HPTLC silica gel 60 plates and subsequent development in toluene-chloroform-diethyl ether-formic acid (20:16:4:0.1, v/v) provided a superior resolution of these compounds.

Restricted access

A selective, precise, and accurate high-performance thin-layer chromatographic (HPTLC) method has been developed for the analysis of Nagod samples and its formulations for their betulinic acid content. The method involves densitometric evaluation of betulinic acid after resolving it by HPTLC on silica gel plates with toluene—acetone—formic acid (17:3:0.2, v/v/v) as the mobile phase. For densitometric evaluation, peak areas were recorded at 445 nm after derivatizing the resolved bands with anisaldehyde—sulphuric acid reagent. The relation between the concentration of betulinic acid and the corresponding peak areas was found to be linear within the range of 200–1200 ng/spot. The method was validated for linearity range, precision, limit of detection (LOD), limit of quantification (LOQ), and accuracy and was demonstrated for quantification of betulinic acid in Vitex negundo L. plant parts and its commercial polyherbal formulations. The method was able to identify and quantify betulinic acid in complex mixtures of phytochemicals and could be extended to marker-based standardization of plant samples, extracts, and its polyherbal formulations.

Restricted access

A new validated high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous determination of anti-inflammatory compounds betulinic acid (BA, 1), 24β-ethylcholesta-5,22E,25-triene-3β-ol (ECTO, 2), and lupeol (LU, 3) in the roots of Clerodendrum phlomidis. Extraction efficiency of marker compounds was studied using normal (cold and hot), ultrasonic, as well as microwave-assisted extraction techniques with various solvents. Well-resolved separation of marker compounds was achieved on silica gel 60F254 plates using the mobile phase consisting of chloroform-methanol (98:2, ½/½). Marker compounds were scanned using the densitometric reflection-absorption mode after post-chromatographic derivatization with vanillin-sulfuric acid reagent. Validation of method was performed as per the International Conference on Harmonization (ICH) guidelines. Report on the occurrence of betulinic acid for the first time in C. phlomidis is of chemotaxonomic importance. In addition, anti-inflammatory potential of the rare sterol ECTO (2) on lipopolysaccharide (LPS)-stimulated production of pro-inflammatory cytokines (tumor necrosis factor-α [TNF-α] and interleukin-6 [IL-6]) was also evaluated as it was not reported earlier.

Restricted access

Betulinic acid (BA), a potent anticancer agent, occurs in more than 50 plant species worldwide, out of which Dillenia and Ziziphus species are found in Gujarat, India. The presented research work was aimed at the development of a single validated high-performance thin-layer chromatographic (HPTLC) method which can quantify BA from different plant species and thereof from different chemical environments. The analysis was performed on a thin-layer chromatographic (TLC) precoated silica gel 60 F254 plate using a mobile phase of toluene-chloroform-ethanol (4:4:1, v/v/v) and after derivatization with anisaldehyde sulfuric acid reagent showed magenta color spot at R F 0.71. Densitometric evaluation of BA was carried out at 525 nm. The suitability of this HPTLC method for quantitative determination of BA was proved by validation in accordance with the requirements of the International Conference on Harmonization (ICH) guidelines. The occurrence of BA is more frequent in bark or stem as compared to leaves. The content of BA was found to be the highest, 0.43%, in Dillenia indica bark.

Restricted access

Triterpenoid betulinic acid (BA) was detected, quantified, and reported for the first time from leaf extract of Achyranthes aspera along with much known oleanolic acid (OA). Extraction was achieved using ultrasonic exposure, and reversed-phase.ultra flow liquid chromatographic (RP.UFLC) technique was employed during investigation. RP.UFLC separation was achieved on a Hibar 250–4.6 mm, 5 μ, Lichrospher 100, C18e column using methanol and water (90:10) as mobile phase with pH adjusted to 5.0 using glacial acetic acid (GAA) in an isocratic mode. The content of BA (0.25 mg g−1 fresh weight [FW]) was ∼75% higher than OA (0.06 mg g−1 FW). These results suggest BA to be the major triterpenoid compared to OA in the leaf of A. aspera. High-performance thin-layer chromatography (HPTLC) separation of the two triterpenic acids (oleanolic and betulinic acid) was also achieved on silica gel G 60 F254, 50 ×~ 100 mm glass TLC plates, using benzene, ethyl acetate, and formic acid as solvent system in a ratio of 67.9:22.7:9.4. Anisaldehyde reagent was used for detection. The method was used for the screening of oleanolic and betulinic acids from A. aspera leaf extract. Similarly, Fourier transform-infrared (FT-IR) spectroscopic analysis was done in the mid IR region of 400–4000 cm−1 with 64 scan speed using OMNIC 8.1 (ver. 8.1.210) software. The results were also supported by HPTLC and FT-IR data.

Restricted access

This article enfolds a rapid and sensitive high-performance thin-layer chromatographic (HPTLC) method for the estimation of four triterpenoids, namely, betulin (BU), betulinic acid (BA), lupeol (LU), and oleanolic acid (OA), from the bark, roots, and leaves of Betula utilis D. Don, an endangered Himalayan tree. All the four phytoconstituents have high therapeutic value. Separation was performed on thin-layer chromatography (TLC) aluminum plates precoated with silica 60 F254 (20 × 20 cm) followed by detection of betulin, lupeol, and oleanolic acid carried out by derivatizing the plate with ceric ammonium sulfate followed by heating at 110°C for 5 min. For betulinic acid, the plate was dried and visualized after spraying with Liebermann‒Burchard reagent. CAMAG TLC Scanner 4 equipped with winCATS software was used for densitometric scanning at 500–550 nm. The proposed technique was further validated in terms of linearity, precision, accuracy, and sensitivity as per the International Conference on Harmonisation (ICH) guidelines. A good linear relationship was obtained for the calibration plots with r 2 = 0.9994, 0.9995, 0.9969, and 0.9998 for betulin, lupeol, oleanolic acid, and betulinic acid, respectively. Accuracy of the method was checked by recovery study conducted at three different levels with the average recovery between 98.9% and 99.3% for all the four markers.

Restricted access

Abstract  

The aim of this study was to examine the radioiodinating condition of betulinic acid and understand the possibility of 131I–betulinic acid (131I–BA) as a potential tumor radiotherapy agent through in vitro uptake and in vivo biodistribution studies 131I–BA was prepared by the reaction of betulinic acid with Na131I in the presence of hydrogen peroxide, and then purified by HPLC. The labeling yield was about 80%, and the radiochemical purity was greater than 95%. 131I–BA was found to be stable at 4 °C in saline containing 1% ethanol. In vitro studies showed that 131I–betulinic acid accumulated in the cancer cell lines (BEL-7402 and NCI-H446) in comparison with free 131I. In vivo biodistribution study in KM mice bearing HepA tumor showed that 131I–BA stayed longer time in tumors than free 131I. A significant differences were seen in tumor/muscle ratio at 4 h postinjection between 131I–BA and free 131I. In vivo and in vitro studies showed the higher fraction of 131I–BA can be utilized for therapy and a higher dose will be delivered per targeting event. 131I–BA is a promising radiopharmaceutical in nuclear medicine, especially for hepatocellular tumor targeted radionuclide brachytherapy.

Restricted access

In view of high pharmacological significance and urgent need to identify high-yielding plant sources of betulin (1) and betulinic acid (2), quantitative high-performance thin-layer chromatographic (HPTLC) analysis has been done on various parts of Diospyros sylvatica. For separation of both compounds, HPTLC silica gel 60 F254 plates were used with chloroform-methanol-formic acid (98:2:2) as the mobile phase. The method was validated for selectivity, sensitivity, accuracy and precision. While betulin was found in maximum amount in stems (0.08%), betulinic acid accumulates maximally in roots (0.29%). The developed HPTLC method is simple, rapid, and highly suitable for the quantification of betulin and betulinic acid in various related species and genera.

Restricted access

A new, simple, reliable, and validated high-performance thin-layer chromatographic (HPTLC) method has been developed for the simultaneous quantitation of four bioactive markers, ursolic acid (1), betulinic acid (2), β-sitosterol (3), and lupeol (4) in the stem and root barks of Alstonia scholaris. Extraction efficiency of the targeted markers from the bark matrixes with organic solvents using cold percolation, hot extraction, and ultrasonic extraction were studied, which showed that ultrasonic extraction was best for sample preparation. The separation was achieved on silica gel 60F254 HPTLC plates using chloroform-methanol (99:1 v/v) as mobile phase. The quantitation of four markers was carried out using the densitometric reflection/absorption mode at 680 nm after post chromatographic derivatization using vanillin-sulphuric acid reagent. The linear regression analysis data for the calibration plots showed a good linear relationship (r 2 ≥ 0.998) in the concentration range 2–10 μg per spot for the ursolic acid and lupeol and 4–20 μg per spot for the betulinic acid and β-sitosterol with respect to area. The method was validated for peak purity, precision, accuracy, robustness, limit of detection (LOD), and quantitation (LOQ). Method specificity was confirmed using retention factor (R F) and visible spectral (post chromatographic scan) correlation of marker compounds in the samples and standard tracks.

Restricted access