Ok, I can only propose a procedure, neither tested, nor used in a laboratory.
I think, this might work when you need to determine amygdaline in the samples:
- weight known amount of the disintegrated sample /dry weight/, add 3% water solution of H2S04 /sample to dilluted acid weight ratio might be 1: 10/, add a half volume of n- hexane or CCl4, and heat the contents at 80 oC for 30 min. under reflux. The addition of organolvent is neccessary from reason of limitted solubility of benzaldhyde in H2O and its evaporation during the hydrolysis.
- after hydrolysis perform at least 2 further extraction steps and combine the extracts.
- neutralize traces of dispersed sulphuric acid in the extract by addition of solid NaHCO3, and remove traces of H2O with molecular sieve or anhydrous Na2SO4.
-reduce volume of the combined extracts by rotavap and analyse it via GLC or HPLC /with use of internal standard or via calibration curve/.
Note: it is on you to find the convenient time and temperature of hydrolysis. Start with the proposed temperature and perform hydrolysis for 10, 20, 30 and 40 min. The optimal time at 8O oC will be that when the amount of benzaldehyde between the two last experiments will be the same. The same could be done also at 70 and 90 oC.
Alkaline hydrolysis was not taken into the consideration due to a number of by-products fomation, incl. those derived from benzaldehyde and degradation products of your samples at high pH and elevated temperature.
Based on the positive preliminary results a modified hydrolysis procedure /optimized/ might be tested in larger scale experiments.
Good luck,
R.S.
Dear Mustapha,
In the case you only need to rid almond or kernel apricot samples of amygdaline, and you are not interested in its quantity /in bitter almond it is 2.5 to 3.5%/, you can do it via hot ethanol Soxhlet extraction of the disintegrated substrates.
R.S.
Mustapha,
beta-glucosidase is an enzyme you need to place an order for. You could get it from Merck, Lambda or other supliers.
Enzymatic hydrolysis however will be rather expensive compared to acidic one. Anyhow, you should use a disintegrated substrate to promote diffusion of enzyme into it, and the time of hydrolysis might be much longer. Temperature might be an ambient one.
Good luck.
R.S.
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