In this experiment, the α-amylase inhibitory activities of the compounds 3a -h and 5a -h were evaluated according to the reported methodology [107 (link)]. Precisely, 10 µL of α-amylase enzyme (3,3 U, EC 3.2.1.1, Sigma Chemical Co., St. Louis, MO, USA) was mixed with 10 µL of each compound at different concentrations ranging from 20 to 200 µg/mL, DMSO or quercetin (used as the positive control at the same compound concentrations) at 37 °C for 5 min. Then, 180 µL of Labtest (the amylase substrate) was added and the samples were incubated for 8 min, followed by measuring the first reaction at 620 nm to obtain (X1). Afterwards, the reaction mixture was incubated for an additional 5 min at 37 °C; then, the second reaction was measured to obtain the final reading (X2). Notably, the amylase substrate, Labtest, was diluted in distilled water (1:1) before being added to the microplate (Bio Tek ELX-800, USA). The α-amylase inhibitory efficiency was calculated as follows: % inhibition = 100 − (X2 sample − X1 sample/ X2 control − X1 control) × 100, where control is the absorbance of the assay with DMSO instead the test compound. The results were expressed as IC50 µg/mL (as well as µM) and were determined from the standard curves.
α amylase enzyme
Manufactured by Merck Group
Sourced in Germany, United States
α-amylase is an enzyme that catalyzes the hydrolysis of α-1,4-glycosidic linkages in starch and other polysaccharides. It is a key component in the breakdown of complex carbohydrates.
Automatically generated - may contain errors
Lab products found in correlation
Acarbose, by Merck Group (8 mentions)
Gallic acid, by Merck Group (4 mentions)
α glucosidase enzyme, by Merck Group (3 mentions)
Ascorbic acid, by Merck Group (3 mentions)
Quercetin, by Merck Group (3 mentions)
Methanol, by Merck Group (3 mentions)
Ethanol, by Merck Group (3 mentions)
Methanol, by Thermo Fisher Scientific (3 mentions)
Elx800, by Agilent Technologies (2 mentions)
Sabouraud dextrose agar, by Thermo Fisher Scientific (2 mentions)
Folin ciocalteu reagent, by Merck Group (2 mentions)
Tween 20, by Merck Group (2 mentions)
3 5 dinitrosalicylic acid, by Merck Group (2 mentions)
Sodium chloride (nacl), by Merck Group (2 mentions)
Folin ciocalteu, by Merck Group (2 mentions)
Phlorizin dehydrate, by Merck Group (2 mentions)
Magnesium chloride 6 hydrate, by Merck Group (2 mentions)
Potassium chloride (kcl), by Merck Group (2 mentions)
α amylase, by Merck Group (2 mentions)
Trichloroacetic acid, by Merck Group (2 mentions)
17 protocols using α amylase enzyme
1
α-Amylase Inhibitory Activity Evaluation
2
Antioxidant and Antimicrobial Evaluation
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Solvents (n-Hexane, chloroform, acetone, ethyl acetate, methanol, ethanol and DMSO), gallic acid, quercetin, aluminium chloride (AlCl3), potassium acetate, 2,2-diphenyl-1-picrylhydrazyl (DPPH), ascorbic acid, sulfuric acid (H2SO4), ammonium molybdate, monosodium dihydrogen phosphate (NaH2PO4), trichloroacetic acid (TCA), potassium ferricyanide, ferric chloride (FeCl3), standard antibiotics (cefixime, ciprofloxacin), standard antifungal (clotrimazole), trypton soy broth (TSB), α-amylase enzyme (from Bacillus subtilis), acarbose, phosphate buffer (PB), Folin–Ciocalteu reagent, RPMI-1640 medium, Medium 119, DMEM and sea salt were purchased from Sigma (Sigma-Aldrich Germany). Sabouraud dextrose agar (SDA) was purchased from Oxoid England; Tween-20 from Merck-Schuchardt, USA. Medium ISP4 was prepared in lab while doxorubicin was purchased from Merck (Darmstadt, Germany).
3
Antioxidant and Enzyme Inhibition Assays
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Merck Company supplied the ethanol. Sigma-Aldrich Company provided the Folin-Ciocalteau-phenol reagent (FC), 1,1 diphenyl-2-picryl-hydrazyl (DPPH), ascorbic acid, and butylated hydroxyl toluene (BHT). Sigma-Aldrich Company supplied α-amylase enzyme and acarbose inhibitor. All the other chemicals were of the highest commercial purity.
4
Comprehensive Biochemical Assay Protocols
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Phosphate phosphate, sodium Chloride, acetone, sodium carbonate were purchased from Acros Organics (USA). Methanol (MeOH), Dithiothreitol (DTT), Ethyline Diamine Tetra Acetic acid (EDTA), 2-propanol, Nitro blue tetrazolium chloride (NBT), Guaiacol, α-Naphthyl acetate solution, Fast blue BB, xylenol orange disodium salt, sulfuric acid (H2SO4), sodium Chloride (NaCl), ferrous ammonium sulphate, glycerol, o-Dianisidine, Potassium acetate, α-amylase enzyme, 3, 5-dinitrosalicylic acid, bovine serum albumin, hydrochloric acid (HCl), hydrogen peroxide H2O2, sodium acetate, Glacial acetic acid, Aluminium chloride AlCl2, sodium phosphate, and 2, 2'-Azino-Bis-3-Ethylbenzothiazoline-6-Sulfonic Acid (ABTs) were purchased from Sigma-Aldrich (Merck Germany). Polyvinyl Pyrrolidone (PVPP), starch and Acetic acid were purchased from Bio-world GeneLinx International, Inc., USA. Folin-Ciocalteu (FC) Reagent, Bradford dye, Sodium Dodecyl Sulfate (SDS) were purchased from Thermo Fisher Scientific UK. 2, 6-dichloroindophenol (DCIP) was purchased from Millipore Sigma US.
5
Carrageenan-Amylase Enzyme Interaction
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κ-carrageenan was obtained from Across Organics, and the α-amylase enzyme (700 U/mL) was purchased from Sigma-Aldrich(St. Louis, MO, USA). RPI Glycerol Liquid, USP grade, and molecular weight (92.1).
6
Biochemical Analysis of Antioxidant Compounds
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Ethanol was purchased from Merck KGaA (Darmstadt, Germany), and gallic acid, Folin-Ciocalteu, streptozotocin, α-amylase enzyme, and acarbose were acquired from Sigma Chemical Co. All reagents were of analytical grade.
7
Diabetes Compounds Characterization Protocol
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Alloxan monohydrate 98%, glucose oxidase-peroxidase reagent, sucrose, starch powder, and magnesium chloride-6-hydrate [MgCl2, 6H2O] were acquired from Sigma-Aldrich, Steinheim, Germany, as for the α-glucosidase enzyme, and acarbose were imported from Sigma-Aldrich, China, furthermore, the α-amylase enzyme and phlorizin dehydrate were both purchased from Sigma-Aldrich, USA, and Pentobarbital was imported from France. Dimethyl-sulfoxide (DMSO), Sodium chloride (NaCl) and potassium chloride (KCl) were purchased from Sigma Aldrich, Riedel-de Haen, Denmark. Ether and glibenclamide have been both acquired from Casablanca and Oujda respectively in Morocco, as for the Calcium chloride dihydrate [CaCl2, 2H2O] was imported from Spain, Scharlau Chemie, s.a. The sodium bicarbonate [NaHCO3] was purchased from Puerto Rico, (Farco Chemical Supply), and finally Sodium phosphate monobasic dihydrate [NaH2PO4 · 2H2O] from Panreac Spain.
8
α-Amylase Inhibition Assay Protocol
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The α-amylase inhibitory activity was determined calorimetrically according to Subramanian et al.43 . Briefly, 4 U of α-amylase enzyme (Sigma, St. Louis, MO) was incubated with 10 μL of each compound (250–31.2 μg/ml), DMSO (negative control), or quercetin (control drug used at the same compound concentrations), at 37 °C for 5 min. Then, samples were mixed with 180 μL of the Amylase Substrate (Labtest®) and incubated for 10 min and the first reaction was measured at 620 nm. Then, 100 μL of the reactive α-amylase (Labtest®) diluted in distilled water (1:1) and 150 μL of distilled water were added in the microplate and then incubated at 37 °C for 10 min and the second reaction was measured again. IC50 values were determined by nonlinear regression. The α-amylase inhibition percentage was determined using the following equation: % inhibition =100 – (A2 sample – X1 sample/X2 control − X1 control) × 100 where X1 is the absorbance of the initial reading and X2 is the absorbance of the final reading.
9
Enzymatic and Salivary Starch Hydrolysis of Rice Granules
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α-Amylase
(240 000 units of the α-Amylase enzyme from Bacillus subtilis, Sigma-Aldrich, Singapore) was
added into the suspension containing 0.01 M phosphate buffer with
pH 7.4 (5 mL) and rice granules (1 g) and stirred at 37 °C; the
treated granules were sampled at 1 and 3 h and then observed by SEM.
In addition, 50 mg of rice granules was dispersed in 5 mL of 0.01
M phosphate buffer with pH 7.4 and 400 μL of fresh saliva was
added and the mixture was incubated at 37 °C with gentle shaking
for 1 and 3 h. In addition, 50 mg of rice granules was dispersed in
5 mL of water and 400 μL of fresh saliva was added and the mixture
was incubated at 37 °C with gentle shaking for 1 and 3 h. A similar
experiment with saliva was carried out using 0.01 M phosphate buffer
with pH 7.4 in place of water.
(240 000 units of the α-Amylase enzyme from Bacillus subtilis, Sigma-Aldrich, Singapore) was
added into the suspension containing 0.01 M phosphate buffer with
pH 7.4 (5 mL) and rice granules (1 g) and stirred at 37 °C; the
treated granules were sampled at 1 and 3 h and then observed by SEM.
In addition, 50 mg of rice granules was dispersed in 5 mL of 0.01
M phosphate buffer with pH 7.4 and 400 μL of fresh saliva was
added and the mixture was incubated at 37 °C with gentle shaking
for 1 and 3 h. In addition, 50 mg of rice granules was dispersed in
5 mL of water and 400 μL of fresh saliva was added and the mixture
was incubated at 37 °C with gentle shaking for 1 and 3 h. A similar
experiment with saliva was carried out using 0.01 M phosphate buffer
with pH 7.4 in place of water.
10
Antioxidant and Enzyme Inhibition Assay
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Safrole oil was purchased from (Sigma-Aldrich, Germany). The carboxyvinyl polymer (Carbopol 940) was obtained from the CBC Co., Ltd., Japan. Dimethyl Sulfoxide (DMSO) was obtained from Riedel-de-haen, (Seelze, Germany). Trolox [(S)-(−)-6-hydroxy-2, 5, 7, 8-tetramethylchroman-2-carboxylic acid] and 2,2-Diphenyl-1-picrylhydrazyl (DPPH) were purchased from Sigma Aldrich (Denmark). Both surfactants Span and Tween were bought from Al-Shamas company (Palestine). DNSA 3,5-Dinitrosalicylic acid (DNSA) reagent was purchased from Sigma-Aldrich (LA, USA) Methanol and n-hexane were obtained from Loba Chemie (India). N-Succ (Ala) 3-o-nitroanilide (SANA) and porcine pancreatic elastase (PPE) were obtained from Sigma Aldrich, USA. The α-amylase enzyme from Sigma Aldrich (Mumbai, India), and acarbose was obtained from Sigma-Aldrich (St. Louis, USA).
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