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Simultaneous densitometric determination of β-sitosterol and lupeol through validated HPTLC method in different plant parts of Uraria picta (Jacq.) Desv. ex DC. – A dashmool species

Saxena Hari Om, Parihar Samiksha, Pawar Ganesh, Rao G. Rajeshwar

Acta Chromatographica

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2023

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Vol. 35, no. 1

99--105

EN

β-sitosterol (BS) and lupeol (LU) exhibit a number of biological activities and are the important bioactive marker compounds in pharmaceutical science. In the present study, a simple, precise, accurate and validated high performance thin layer chromatographic (HPTLC) method was developed for simultaneous quantification of these two compounds in leaves, stem and roots of Uraria picta, a critically endangered medicinal plant and one of the important constituents of ten plants ayurvedic formulation called Dashmoola. Standards and test samples were applied on TLC aluminum plate precoated with 0.2 mm layer of silica gel 60F254. The plate was run in a twin glass chamber comprising toluene: methanol: chloroform (8:1:1, v/v/v) as a mobile phase. The plates were immersed in anisaldehyde-sulfuric reagent and then heated at 105 °C for 5 min in CAMAG TLC plate heater for appearance of bands. Densitometric scanning was performed at λmax = 525 nm using tungsten light source in CAMAG TLC Scanner4 armed with WinCATS software. RF values of BS and LU standards and those of test samples were found to be 0.53 ± 0.01 and 0.63 ± 0.01 respectively. The method was further validated by following the International Conference of Harmonization (ICH) guidelines. For BS and LU, the linear regression data for the calibration plots revealed a satisfactory linear association with r2 = 0.995 and 0.998, respectively. Linear range for both BS and LU was 200–600 ng/band. Accuracy of the method was evaluated by performing recovery study at three different levels by standard addition methods with an average recovery of 99.86% and 101.07%. The results revealed that the leaf samples of U. picta contained highest concentration of BS (0.150 ± 0.02%) while its root samples confined the highest concentration of LU (0.149 ± 0.01%). The developed method can be applied for routine and quality control analysis in different herbal formulations containing U. picta species.

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Chromatographic identification of two biologically important triterpenoids from the chloroform extract of Rhizophora mucronata

Hridya V. K, Godson P. S, Chandrasekar N.

Acta Chromatographica

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2012

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Vol. 24, no. 1

123--129

EN

Mangroves are the source of several bioactive secondary metabolites and have proven activity against human, animal and plant pathogens. Rhizophora mucronata is a mangrove species belonging to the family Rhizophoraceae. Betulin and lupeol are two naturally occurring pentacyclic triterpenoids having excellent biological properties. Chloroform extract of R. mucronata, collected from Pitchavaram, Muthupett and Manakudy regions of Tamilnadu, was chromatographed on silica gel 60F254 plates with nhexane and ethyl acetate (80:20) (v/v) as the mobile phase. Derivatizations were done using anisaldehyde and sulfuric acid reagents. The triterpenoids such as betulin and lupeol were identified through HPTLC method. This is the first report for the thin layer chromatographic (TLC) identification of triterpenoids such as betulin and lupeol from R. mucronata, which will be the most valuable information to identify the antimalarial and antiviral activities.

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Development and validation of a method for densitometric analysis of lupeol from Mimosoups elengi

Ganu G. P, Jadhav S. S, Deshpande A. D

Acta Chromatographica

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2010

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Vol. 22, no. 3

491--497

EN

In this paper we describe a simple, precise, and accurate HPTLC method for quantification of lupeol in the bark of Mimosoups elengi (ME). The bark was extracted with hot methanol in a Soxhlet apparatus. Chromatographic separation of the extract was performed on aluminum foil plates coated with silica gel 60 F 254 as the stationary phase. The mobile phase was toluene-ethyl acetate-formic acid 12:2:1 ( υ/υ ). Densitometric evaluation of the separated zones was performed at 220 nm. The lupeol was satisfactorily resolved at R F 0.64 ± 0.02. The accuracy and reliability of the method were assessed by evaluation of linearity (1000–4000 ng per band), precision (method and instrumental precision, as RSD, 1.06 and 1.03%, respectively), accuracy (97.28 ± 0.89), and specificity in accordance with ICH guidelines.

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Triterpenes in traditional and supercritical-fluid extracts of Morus alba leaf and stem bark

Böszörményi A, Szarka Sz., Héthelyi É, Gyurján I, László M, Simándi B, Szőke É, Lemberkovics É

Acta Chromatographica

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2009

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Vol. 21, no. 4

659--669

EN

Our objectives were to establish a GC method capable of quantitative analysis of terpenoids without derivatisation and to examine the amount of β -sitosterol extracted from Morus alba L. leaf and stem bark by use of traditional organic solvent extraction and supercritical-fluid extraction (SFE). To measure β -sitosterol content without derivatization, GC-FID was used with 5- α -cholestan-3-one as internal standard. To identify terpenoid constituents, GC-MS was used; β -sitosterol, phytol, lanost-7-en-3-on, α -amyrin, β -amyrin, and lupeol were identified. We established that for Morus leaf the best SFE method for β -sitosterol was pilot scale SFE; the β -sitosterol content of this extract was higher than that of the hexane solvent extract. Among analytical SFE conditions, 200 bar for 90 min and 300 bar for 60 min resulted in extraction of the most β -sitosterol. For mulberry stem bark, solvent extraction with hexane and SFE at 400 bar and 40°C for 60 min proved the best methods.