hey guys! since you guys are sharing so much regarding about your SIP stuffs, guess i should not be too stingy with mine too! Just the other day i was in the lab performing carbohydrate analysis, i realized that alpha-amylase is actually an endoenzyme that cleaves both amylose and amylopectin molecules internally and it acts on the (1-> 4) linkages of the starch and attacks the uncooked and undamaged starch granules.

In addition, in order to get the analysis started, we must first ensure that all the equipment are in place and that the sample that we are working on retains as much moisture as possible in order to ensure the results to be as close as the outside analysis as possible. In order to make the sample dry, we put the sample into a vacuum oven overnight. however, i cant  take down the picture cause im not too sure if it will affect TP. after which, we grind the sample into less than 50mm, after which, we put the sample into a duran bottle together with a buffer by the name of MES-TRIS. during this step, the measurement of the sample must be very accurate. after this step, we added in a magnetic stirrer and place it on a magnetic plate and put in 50 microlitre of the alpha amylase that i have mentioned earlier. after which, we placed all the bottles in to a water bath at 80 degrees with an incubation with continuous agitation. after which, we will cool the bottles by replacing the hot water in the water bath to cool water at about 60 degree before adding 100 microlitre of protease. after adding the protease, the bottles are being placed in the water bath to incubate for 30 minutes with continuous agitation.

the next step will be the pH adjustment. as we take the samples out, we added in 5mL of 3M acetic acid into the bottles one at a time to prevent evaporation loss. be adding drop by drop, all the results are being very precise. if the pH is more than 8.0, we would have to do the entire experiment aain. immediately, we added 200 microlitre of amyloglucosidase solution and incubate it again for 30 minutes at 60 degrees with continuous agitation. after which, we transferred the sample into a cuvette that is later being put into a UV spectrophotometer for checking for the absorbance of the product. after the first test, solution A1 is added and we waited for awhile before the second reading. after the second reading, solution A2 was added and we waited for awhile before the readings start. lastly, we put in solution A3 and waited for awhile before the last reading was being read.

this was what i have done in the past week(: hope you guys have a better understanding at carbohydrate analysis(:
-FM(: