Im doing alpha glucosidase inhibitory assay for acarbose which extracted from the acarbose tablets. But i got completely opposite results for the inhibitory activity.
Which mean, with increasing the acarbose concentration, absorbance values are increasing ( the inhibitory activity is decreasing ) what is the reason for this?
1. 25 micro liters phosphate buffer(30mM, pH6.5)
2. 40 micro liters acarbose
3. 40 micro liters enzyme ( 4u/ml)
4. Incubate at 40 c for 15 min
5. 100 micro liters of pNPG
6. Incubate 1 h at 40 c
7 absorbance was measured at 405 nm in microwell plate reader
I got good absorbance value for the negative control which contains buffer instead of acarbose.
It would be greatful if you could help me.
hi,
Acarbose reversibly inhibits intestinal alpha-glucosidases, enzymes responsible for the metabolism of complex carbohydrates into absorbable monosaccharide units.Acarbose is a pseudotetrasaccharide, a natural microbial product derived from cultures of Actinoplanes strain SE 50. This medication inhibits alpha-glucosidases in the brush border of the small intestine, delaying glucose absorption by decreasing the breakdown of complex carbohydrates.Compared with placebo, α-glucosidase inhibitors have minimal effects on body weight. As predicted from their mechanism of action, hypoglycaemic adverse effects do not occur. Abdominal discomfort (flatulence, diarrhoea and stomachache) are the most frequently occurring adverse effects inhibitors and are dose-related.
The inhibition of α-amylase by acarbose led to undigested starch into the large intestine. The colonic fermentation of the undigested starch was regarded as the main cause of side effects of acarbose, including gastrointestinal upset, flatulence, and abdominal bloating.
for more info:
https://www.mdpi.com/1420-3049/27/3/639/pdf
best wishes..
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