I worked on two related plants. I want to identify that both plants contain the same secondary metabolites by taking a single metabolites.
The quantification of certain plant ,aterials such as swertiamarin, mangiferin and amarogentin was carried by reverse phase High Performance Liquid
Chromatography (HPLC Waters 515). For this purpose plant material is grounded separately in liquid nitrogen and suspended in 80% methanol. The sample mixture is sonicated for 10 min. Following day, centrifuge the samples at 10,000 rpm for 10-15 min. and the supernatant is filtered through 0.22μ filter. HPLC method is used to optimize the rate and simultaneous quantification of major three
metabolites.The plant samples are extracted with 80% methanol in a
bath sonication for rapid extraction of metabolites. The desired separation and baseline is obtained in a gradient method where mobile phase A was composed of 0.1% TFA in water and mobile phase B was a mixture of acetonitrile: water in the ratio of 70:30. The linear gradient at a flow rate of 1.0ml/min start with 15% B; 20% B in next 5 min, 70% B in next 25 min, hold for 5 min; 15% B in next 5 and equilibrated for 5 min at 240 nm UV wavelength. The separation was obtained on Waters Spherisorb 5µm ODS2 (250 mm x 4.6 mm). The compounds were identified on the basis of their retention time and comparison of UV spectra with the authentic standards procured from ChromaDex Inc. present. High-Performance Thin-layer Chromatography. The separation can performed on TLC aluminium plates precoated with silica gel 60 F 254 . Goodseparation can be achieved in the mobile phase of Ethyl acetate: Formic acid: Glacial acetic acid: Water 100:11:11:26,v/v) and densitometric determination of these compounds was carried out at 366 nm in reflection/absorbance mode.The rutin and quercetin content of hydroalcohol bark extract of Anogeissus latifolia are found to be 0.1617% w/wand 1.875%w/w respectively.
Please see the attached paper. They also involved the use of HPTLC for the analysis of the in vivo and in vitro plant materials.
"Subcellular Localization and Quantification of Camptothecin in Different Plant Parts of Chonemorpha Fragrans"
[The objective of the present study was to reveal the accumulation of the alkaloid camptothecin (CPT) in both in vitro as well as in vivo plant material of C. fragrans.]
Dear Jitendriya, you may have a look into the folioing (attached link) article related to your query.
Article In vitro accelerated mass propagation and ex vitro evaluatio...
Dear Jitendriya, here is other few articles in this aspect. Find the links below. Good luck!
http://www.sciencedirect.com/science/article/pii/S0102695X15002215
Article Comparative HPTLC Analysis of Antioxidant Compound Gallic Ac...
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