I took 5 mg of extracted PHA and added methanol (with 7% sulphuric acid) and then chloroform followed by incubation at 100 degrees (waterbath) for 3 hrs in screw capped bottles. But the problem I am facing is that its getting evaporated and if the bottle is tightly closed the cap burst. so I have to add the solution two three times again in between. And after incubation when I add water, it becomes turbid and the chloroform phase was clear. I took the bottom phase (chloroform) and run it in GC, I am getting only chloroform peak and nothing else. please suggest me what should I do.
Hi.
From what you say, you are following a sort of "standard procedure" to analyse PHA. For sure you must change reaction vessels. Don't forget that the esters forming are volatile and if you the vessel is not sealed, then many will be lost. Also, take a look at the article bellow. It provides good insight and also refers to other relevant works.
Article Oehmen A, Keller-Lehmann B, Zeng RJ, Yuan ZG, Keller E.. Opt...
Cheers
Thankyou Mario Silva Sir. I will definitely follow the paper you shared.
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