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EN
A new, simple and reproducible HPTLC-densitometric method has been established and validated for estimation of trigonelline in leaves of Abrus precatorius L. and its herbal formulation Chatak® using ICH guidelines. Accelerated solvent extraction (ASE) as an alternative to convention techniques was also explored for the rapid extraction. The methanol extracts of leaves, its formulation, and standard solution of trigonelline were applied on silica gel F254 HPTLC plates and developed in twin chamber using mobile phase toluene-ethyl acetate-formic acid-methanol (2:3:1.8:3.8, v/v/v/v). The plates were scanned at 268 nm (λmax of trigonelline) using Camag TLC scanner 3 with CATS 4 software. A linear relationship was obtained between response (peak area) and amount of trigonelline in the range 40–200 ng spot−1; the correlation coefficient was 0.9957. ASE method has higher extraction efficiency in less time as compared to Soxhlet extraction. The HPTLC-densitometric method showed good linearity, recovery and high precision of compound. It is useful to analyze trigonelline in A. precatorius leaves and routine quality control of its marketed formulation.
EN
A new, simple, and reproducible HPTLC-densitometric method has been established and validated for estimation of trigonelline in the leaves of Abrus precatorius L. and its herbal formulation Chatak® using ICH guidelines. Accelerated solvent extraction (ASE) as an alternative to conventional techniques was also explored for rapid extraction. The methanol extracts of leaves, its formulation, and standard solution of trigonelline were applied on silica gel F254 HPTLC plates and developed in a twin chamber using the mobile phase toluene-ethyl acetate-formic acid-methanol (2:3:1.8:3.8, υ/υ/υ/υ). The plates were scanned at 268 nm (λmax of trigonelline) using a Camag TLC scanner 3 with the CATS 4 software. A linear relationship was obtained between the response (peak area) and the amount of trigonelline in the range 40–200 ng per spot; the correlation coefficient was 0.9957. The ASE method gives higher extraction efficiency in less time compared to Soxhlet extraction. The HPTLC-densitometric method showed good linearity, recovery, and high precision. It is useful to analyze trigonelline in A. precatorius leaves and routine quality control of its marketed formulation.
EN
A simple, sensitive, and reproducible high-performance thin-layer chromatographic method has been established for simultaneous analysis of trigonelline and 4-hydroxyisoleucine from fenugreek seeds ( Trigonella foenum-graceum ). Chromatography was performed on silica gel GF 254 with n -butanol-methanol-acetic acid-water 4:1.5:1:1 (υ/υ) as mobile phase. Densitometric scanning of the plates at 266 nm was used for analysis of trigonelline, and plates were scanned at 395 nm after derivatisation with ninhydrin reagent for analysis of 4-hydroxyisoleucine. The method was validated for specificity, precision (intraday and interday), accuracy, and robustness. Response was a linear function of the concentration of standard solutions in the range 100 to 1000 ng per band and 50 to 500 ng per band, respectively, with correlation coefficients of 0.9992 š 0.04 and 0.9986 š 0.06, for trigonelline and 4-hydroxyisoleucine. The accuracy of the method was checked by determination of recovery at three different levels; average percentage recovery was 99.39 š 0.34 for trigonelline and 99.03 š 0.26 for 4-hydroxyisoleucine. The method was found to be simple, precise, specific, sensitive, and accurate and can be used for routine quality control of fenugreek seeds and also for quantitative analysis of these marker compounds in processed fenugreek and its marketed preparations.
EN
The 1:1 and 2:1 complexes of trigonelline (TRG) with HBr, HI, HNO3, HClO4 and HBF4 have been synthesized and their FTIR, 1H and 13C NMR spectra were analyzed. The 1:1 complexes are divided into two groups. The first group includes complexes with HBr,HI andHNO3, in which a proton is transferred from the acid to TRG and the anions are bonded with the carboxylic group, COOHźźźX- (1). The second comprises complexes with HClO4 and HBF4, with two molecules of protonated TRG forming dimer, like benzoic acid, and the anions interacting exclusively with the positively charged nitrogen atoms (2). All 2:1 complexes crystallized with one water molecule and their FTIR spectra are of Had4i type iii, characterized by an intense broad (continuum) absorption below 1600 cm-1 typical of a short-strong hydrogen bond (SSHB) with a delocalized proton and a single _C=O band. In these complexes the water molecule interacts electrostatically with one positively charged nitrogen atom and the anions with the second one (3). The water molecule additionally forms a hydrogen bond with Br, I and NO3 ions. The proton and carbon chemical shifts of the complexes in D2O have been determined and discussed.
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