For the detection of furosine compound in rice, I have followed several steps,
700 mg rice flour sample hydrolyzed with 8 M of 8 ml HCl solution ,for 23 h at 110 °C . Then I have filtered and found 3 ml solution from here. From this 3 ml I have taken 1ml solution and pass with the C18 cartidge filter for reverse phase HPLC and elluted with 3 M of 3 ml HCl solution. Then the final volume was 4 ml. From this 4 ml final solution I took 1 ml solution to run HPLC. My standard curve result is, y = 242.35x, R² = 0.9987. My compound Retention time is 6.67 and Area is 774, height 59 and wedth is 0.1784. Now how should I calculate to determine the furosine amount (In ppm) present in my sample???
In my 1000 mg of rice flour the amount of protein is 75 mg.
Md Arif Hossain please share the protocol for HPLC. I'm beginner in this field.
Dear Nilofar Nilofar , HPLC protocol is not fixed. It will be depended on your desire compounds. Like if you want to determine the polyphenols from leaves extract, you have to follow the related reference papers. Select some latest and updated papers, try to avoid 5 years old papers. Then compare all selected papers similarities and differences and finally select a latest one. At first run HPLC following by their protocol, sample preparation, gradient program, mobile phase %, flow rate, temperature, wave length etc. Then change every parameter ( flow rate, injection volume, temperature, solvent flow % of A & B) one by one to find clear and separated peak. But at first you have to run your Standard to determine your compound retention time and then Sample. Though I am not expert, you can take my poor knowledge to your consideration whatever are receivable.
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