1. Screening for Antidiabetic Activity of Selected Medicinal Plants

2. Invitro antidiabetic assays (Faiyaz ahmed et.al, 2009)
1.α-amylase inhibition assay
Sample (1%) + 25mL potato starch (4%)
+100mg α-amylase
Vigorous shaking, incubation, 37°C/60 mins
+0.1M NaOH
Centrifuge 3000rpm/15min
Supernatant
Glucose estimation - GOD/POD

3. 2.α-glucosidase and sucrase activity Preparation of enzyme solution
Wistar rats, gms (fasted overnight)
sacrificing by cervical dislocation
intestine excision
washing with ice cold maleate buffer (pH6, 0.1M)
brush border removal
homogenisation with maleate buffer (1:5)
centrifuge, (10,000rpm/10mins)
supernatant
crude enzyme source

4. α-Glucosidase assay Sucrase assay
Enzyme solution (10µL) + sample(10µL)
200µL maleate buffer (pH6.0)
Incubation 10min/37°C
+200µL PNPDGP(2mM)
Incubation 30min/37°C
Boiling water bath, 5min
1mL disodium hydrogen phosphate (0.1M)
Absorbance, 400nm
Enzyme solution (10µL) + sample(10µL)
180µL maleate buffer (pH6.0)
Incubation 10min/37°C
+100µLsucrose solution (60mM)
Incubation 30min/37°C
+200µL 3,5,-DNS
Boiling water bath, 5 min
Absorbance, 540nm

5. Extract + 1mL glucose solution (5-25mM)
3.Glucose uptake by yeast cells
Commercial baker’s yeast - repeated washing in distilled water by centrifuging ( 3000rpm/5min) - until supernatant was clear
10% of the suspension using distilled water.
Extract + 1mL glucose solution (5-25mM)
Incubation 10min/37°C
+100µL yeast suspension
Vortex
Incubation 60min/37°
Centrifuge, 2500rpm/5min
Supernatant
Glucose estimation