Search Results

You are looking at 1 - 9 of 9 items for

  • Author or Editor: Kedar Rout x
  • All content x
Clear All Modify Search

An efficient, sensitive, and precise high-performance thin-layer chromatographic (HPTLC) method has been established for analysis of 6-gingerol in marketed Ayurvedic formulations and in the rhizomes of different varieties of Zingiber officinale . HPTLC separation was performed on aluminum foil-backed HPTLC plates coated with 0.2-mm layers of silica gel 60 F 254 , with n -hexane-acetone, 7.2:2.8 ( v/v ) as mobile phase. Plates were developed to a distance of 78 mm at 20 ± 4°C in a chamber previously saturated for 4 min. Under these condition the retention factor ( R F ) of 6-gingerol was 0.23 and the compound was quantified at 286 nm, its wavelength of maximum absorbance. The limits of detection and quantification were 40 and 150 ng per band, respectively. Response to 6-gingerol was a linear function of amount over the range 150 to 900 ng per band; the correlation coefficient was 0.9997, indicating a good relationship between peak area and amount. Recovery from 98.46 to 101.11% showed the accuracy of the method was excellent. The method is very accurate, simple, and cost effective, and enables sensitive quantitative analysis of 6-gingerol.

Restricted access

HPTLC has been used for quantitative analysis of aloin, as marker compound, in commercial Ayurvedic preparations, in the solid dosage form, containing Aloe vera Linn as major herbal ingredient. Chromatography was performed on aluminum plates coated with 0.2-mm layers of silica gel 60 F 254 , with ethyl acetate-methanol-water 10:1.4:1 ( v/v ) as ternary mobile phase after saturation at 30 ± 4°C for 5–7 min. The development distance was 70 mm. Aloin was quantified at 360 nm, its wave length of maximum absorbance. The limits of detection and quantification were 10 and 20 ng per band, respectively. Regression analysis of the calibration data revealed a good linear relationship ( r = 0.9998) between peak area response and concentration in the range 20 to 100 ng per band. Instrumental precision and the repeatability of the method were 0.44% and 0.89% (CV), respectively. The accuracy of the method, determined by measurement of recovery at three different levels, was in the range 98.13 to 99.75%. These results are indicative of the excellent reliability, reproducibility, accuracy, and precision of the method. The method has been successfully used for analysis of aloin in commercial formulations containing Aloe vera .

Restricted access

A simple, sensitive, and rapid high-performance thin layer chromatographic (HPTLC) method has been established for estimation of piperine in commercial Ayurvedic formulations and in the fruits of Piper nigrum Linn, and Piper longum Linn. Chromatography was performed on aluminum foil HPTLC plates coated with 0.2 mm layers of silica gel F 254 , with hexane-acetone 6.5: 3.5 ( v/v ) as mobile phase. The development distance was 76 mm, the temperature 25 ± 5°C, and the chamber was saturated for 5 min. Piperine was quantified at 340 nm, its wavelength of maximum absorbance. Under the conditions used the R F of piperine was 0.33 and the limit of detection (LOD) was 4 ng per zone. The calibration plot was linear in the range of 10 to 60 ng per zone with a correlation coefficient of 0.9996. Recovery was in the range 98.76 to 100.70%. This HPTLC method was found to be reproducible, accurate, and precise and could be used to detect piperine at nanogram levels. The method is a very simple and cost-effective means of quantitative estimation of piperine in Ayurvedic formulations.

Restricted access

Lupenone was isolated for the first time from the stem bark of Diospyros melanoxylon and characterized with the help of physical and spectroscopic data. A high-performance thin-layer chromatography (HPTLC) method has been developed and validated for the quantification of lupenone in D. melanoxylon stem bark. HPTLC analysis was performed on HPTLC plates by using a binary mobile phase of n-hexane–ethyl acetate (8.2:1.8, v/v). It was quantified at 395 nm after derivatization with methanol—sulfuric acid reagent. The limits of detection and quantification were found to be 40 ng and 100 ng per spot, respectively. The linear regression analysis data for the calibration plot in the range of 100–500 ng spot−1 showed a good linear relationship between peak area and concentration (r 2 = 0.9997). The instrumental precision was 1.01% (coefficient of variation [CV]) and the repeatability of the method was 2.17% (CV). The method was validated for precision, recovery, and repeatability as per the International Conference on Harmonization guidelines. The developed HPTLC method is very accurate and precise and has been successfully used for the estimation of lupenone in D. melanoxylon stem bark.

Restricted access

A high-performance thin-layer chromatography (HPTLC) method has been developed and validated for the quantification of two bioactive lupane triterpenoids, namely, lupeol and betulin from Diospyros melanoxyon stem bark. Chromatographic separation was achieved on aluminium foil-backed HPTLC plates using ethyl acetate-hexane (1.8:8.2, v/v) as mobile phase. The compounds were quantified at their wave length of maximum absorbance in the range of 100–500 ng per spot. The instrumental precision was 0.82% and 1.07% (CV) and the repeatability of the method was 1.33% and 1.17% (CV), respectively, for lupeol and betulin. The minimum detectable amount was found to be 40 and 50 ng per spot for lupeol and betulin, respectively. The linear regression analysis data for the calibration plots showed a good linear relationship with r 2 = 0.9996 for lupeol and 0.9997 for betulin. The method was validated for precision, recovery, and repeatability as per the International Conference on Harmonization guidelines. The developed HPTLC method is very accurate and precise, and has been successfully applied for the assay of these bioactive molecules in D. melanoxylon stem bark.

Restricted access

An important bioactive molecule, ursolic acid was isolated from the leaves of Diospyros melanoxylon and characterized with help of physical and spectroscopic data viz. m. p, IR, 1H, and 13C NMR. A high-performance thin-layer chromatography method has also been developed and validated for its quantification in D. melanoxylon leaves. The high-performance thin-layer chromatography analysis was performed on high performance thin-layer chromatography plates using chloroform-methanol (9.5:0.5, v/v) as mobile phase. The compound was quantified at 540 nm after derivatzation with sulphuric acid reagent. The sensitivity of the method with respect to limit of detection and limit of quantification were found to be 20 and 40 ng per spot. The response was obtained as a linear function of peak area and concentration in the range of 50 to 450 ng per spot with correlation coefficient of r 2 = 0.9998. The method showed excellent accuracy greater than 97.54% with acceptable precession and was successfully validated according to International Conference of Harmonization protocols. Antimicrobial screenings of ursolic acid revealed potent activity against two Gram-positive bacteria viz. Staphylococcus aureus and Enterococcus faecalis and three fungal starins viz. Aspergillus niger, Candida tropicalis, and Candida albicans.

Restricted access

A simple normal-phase high-performance thin-layer chromatography (HPTLC) method has been developed for the quantification of β-sitosterol in hairy root cultures of Clitoria ternatea vis-à-vis natural plant parts. Chromatographic separation was achieved on normal-phase TLC plates by using optimized mobile phase of n-hexane. acetone (8:2, v/v). Densitometric scanning was performed after postchromatographic derivatization of the plate at 414 nm. The limits of detection and quantification were found to be 40 ng and 100 ng spot−1, respectively. A linear response of calibration line was observed over the range of 100 to 500 ng spot−1 with a correlation coefficient of r 2 = 0.999. Recovery values from 97.22 to 98.51% with small coefficient of variation showed excellent accuracy of the method. The method was validated according to the International Conference on Harmonization (ICH) protocol. The method enables excellent separation and accurate quantification of β-sitosterol in different plant parts of the two floral varieties of C. ternatea along with their cultured roots derived through genetic transformation. β-Sitosterol content was comparable between the blue-flowered and white-flowered plants; roots had a higher content than that in stem and leaf regardless of the two floral varieties. Transformed root cultures had a lower content of β-sitosterol compared to natural roots.

Restricted access

The bioactive molecule, α-amyrin acetate, was isolated from the nhexane extract Streblus asper stem bark by column chromatography. The structure of the compound was characterized with the help of physical and spectroscopic data. A high-performance thin-layer chromatography (HPTLC) method has been developed and validated for its quantification in S. asper leaf, stem bark, and root. Chromatographic separation of the compound was achieved on high-performance precoated TLC plates by using optimized binary mobile phase consisting of n-hexane-ethyl acetate (9.6:0.4, v/v). HPTLC scanning was performed at 472 nm after derivatization of the plate with methanol-sulphuric acid reagent in reflection/absorption mode. The method was validated according to International Conference on Harmonization (ICH) protocol for specificity, sensitivity, linearity, accuracy, precision, repeatability, and robustness. The developed method is found to be very simple, rapid, precise, sensitive, and accurate for the quantification of α-amyrin acetate in S. asper.

Restricted access

A simple, sensitive, and precised high-performance thin-layer chromatography (HPTLC) method has been developed for the analysis of lawsone in natural vis-à-vis micropropagated plant parts of Lawsonia inermis L. Separation of the components was perfectly achieved on high-performance thin-layer chromatography (TLC) plates using optimized tertiary mobile phase of benzene‒ethyl acetate‒ acetic acid (7.5:2.5:0.1, v/v). Densitometric scanning was performed before derivatization of the plate in absorption/reflection mode, and lawsone was quantified at its maximum absorbance of wavelength of 275 nm. Linearity of the method was obtained in the concentration range of 50 to 350 ng spot−1 with a correlation coefficient (r 2) of 0.9999, indicating good relationship between concentrations in opposition to the peak area. The limit of detection and limit of quantification were found to be 16 and 50 ng spot−1, respectively. The obtained recovery ranges from 95.09% to 96.90% with an average value of 96.02% proved the excellent accuracy of the method. The developed method was found to be highly sensitive, and the mobile phase enables outstanding separation of lawsone from other components present in the mixture. The International Conference on Harmonization (ICH) guidelines were followed for validation of the HPTLC method in terms of precision, repeatability, and accuracy. The maximum content of lawsone was reported in the leaves of the micropropagated plant.

Restricted access