Caffeic acid, ferulic acid, 3,4-dimethoxycinnamic acid, 5-caffeoyquinic acid, 3,5-dinitrosalicylic acid, potassium sodium tartrate, amylose and human salivary α-amylase type IX-A were all purchased from Sigma-Aldrich. Co., Ltd., Dorset, UK. Oasis MAX cartridge 1 mL (30 mg) and 3 mL (60 mg) were purchased from Waters Ltd., Milford, MA, U.S.A. All the reagents were of the highest purity and standards were ≥98 %. The colour reagent was prepared by mixing 20 mL of 96 mM of 3,5-dinitrosalicylic acid with 8 mL of 5.3 M (12 g in 8 mL of 2 M sodium hydroxide) and 12 mL Millipore water. Human salivary amylase type IX-A stock concentration of 1.25 U/mL was prepared in PBS (0.01 M, pH 6.9) to give 0.5 U/mL in the assay according to the optimized assay (Nyambe-Silavwe et al., 2015 ).
Human salivary α amylase type 9 a
Manufactured by Merck Group
Sourced in Spain
Human salivary α-amylase type IX-A is a laboratory reagent used in research applications. It is an enzyme that hydrolyzes α-1,4-glucosidic linkages in polysaccharides, such as starch and glycogen.
Automatically generated - may contain errors
Lab products found in correlation
Sodium potassium tartrate, by Merck Group (1 mentions)
Oasis max cartridge, by Waters Corporation (1 mentions)
3 4 dimethoxycinnamic acid, by Merck Group (1 mentions)
3 5 dinitrosalicylic acid, by Merck Group (1 mentions)
Caffeic acid, by Merck Group (1 mentions)
Ferulic acid, by Merck Group (1 mentions)
Pefabloc, by Merck Group (1 mentions)
Pancreatin, by Merck Group (1 mentions)
2 protocols using human salivary α amylase type 9 a
1
Enzymology Protocol for Amylase Activity
2
In Vitro Digestion of Food Samples
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MA, M and SM were digested as previously reported (Minekus et al., 2014) . Briefly, samples (3g) were digested in simulated salivary fluid (pH 7.0) containing 75 U/mL human salivary αamylase type IX-A (EC 3.2.1.1, Sigma-Aldrich, Madrid, Spain) for 2 min at 37 °C. Subsequently, gastric fluid (pH 3.0) containing 2000 U/mL pepsin (EC 3.4.23.1, was added and incubated at 37 °C in an orbital shaker at 150 rpm for 120 min. Afterwards, simulated intestinal fluid containing 100 U trypsin activity/mL pancreatin (Sigma-Aldrich) and bile extract (10 mM in the final mixture) is added and incubated at 37 °C in an orbital shaker at 150 rpm for 120 min.
Digestions were performed in duplicate. Sample aliquots were withdrawn at the starting point (negative control) and after gastric (120 min) and intestinal digestion (240 min) and enzymatic activity was immediately inactivated using 5 mmol/L Pefabloc (Sigma-Aldrich). All samples were freeze-dried and stored at -20 °C until analysis.
Digestions were performed in duplicate. Sample aliquots were withdrawn at the starting point (negative control) and after gastric (120 min) and intestinal digestion (240 min) and enzymatic activity was immediately inactivated using 5 mmol/L Pefabloc (Sigma-Aldrich). All samples were freeze-dried and stored at -20 °C until analysis.
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