I want to quantify the jasmonic acid (JA) content in plant using HPTLC. I am following the method described by Dhandhukia and Thakkar, 2008. J. of Chromatographic Science, 46:320-324. After the separation on the TLC plate, the JA band were not visible at 254 nm. I had also tried to stain the JA using Sulphuric acid as reported in some paper, but that also did not produce any band. What is the best option for staining JA on the TLC plate.
I have only abstract: Maybe you can find the complete paper
O. MIERSCH*, G. SEMBDNER, K. SCHREIBER, (*Inst. of Plant Biochem., Acad. of Sci. of the GDR, 4050 Halle (Saale), Weinberg 3, GDR): Occurrence of jasmonic acid analogues in Vicia faba. Phytochemistry 28, 339-340 (1989). TLC of (-)-9,10-dihydro-jasmonic acid, 3,7,-didehydrojasmonic acid and (+)-6-epi-7-iso-cucurbic acid on silica with hexane - ethyl acetate - acetone 60:40:1 and chloroform - methanol - water 140:20:1. Detection with anisaldehyde reagent and heating for 5-10 min. at 120°C under UV 254 nm.
To Geetgovind Sinam,
2,4-DNP strain can also be used for the detection of aldehydes and ketones. In jasmonic acid, there is a keto group, so the 2,4-DNP strain will be very much useful to detect. For preparing the 2,4-DNP, Dissolve 12 g of 2,4-dinitrophenylhydrazine, 60 mL of H2SO4, and 80 mL of H2O in 200 mL, 95% EtOH.
Ref: J Chromatogr Sci. 2008 Apr;46(4):320-4. Separation and quantitation of jasmonic acid using HPTLC. Dhandhukia PC, Thakkar VR.
To Elancheran, Thank you for your reply. Can you also tell the procedure on how to stain Jasmonic acid using 2,4-DNP.
First, you prepare the 2,4-DNP solution. then do TLC in TLC Silica gel 60 F254 plates in the optimized solvent system. then spray the 2,4-DNP solution or dip the TLC plate in the solution, Heat at 60-80 oC, If the spot turns to orange colour, it will confirm the aldehyde or keto groups.
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