Glucosidase hydrolyzes the substrate mix to release the pnitrophenol that can be measured colorimetrically od 410 nm. There is only one tablet of this type used, called acarbose. Alphaglucosidase and alphaamylase inhibitory activities. The modes of inhibition of both enzymes was determined using lineweaverburke plot. Where y is ic 50 50, m is the gradient of the curve, x is the concentration and c is the intercept at y axis 2. This is evident from the intersected lines at the x. Alpha glucosidase inhibitor it is usually used when a healthy diet and physical activity alone has been unsuccessful, although it is sometimes used together with a sulphonylurea. The ability of berry extracts to inhibit alphaglucosidase in. In a 96well microplate, each well was added with 10 l of extract dissolved in dmso and 90 l of 0. Effective control of postprandial glucose level through inhibition of intestinal alpha glucosidase by cymbopogon martinii roxb.
The alphaglucosidase inhibitory and antioxidant activities of r. One mg of glucosidase saccharomyces cerevisiae, sigmaaldrich, usa was dissolved in 100 ml of phosphate buffer ph 6. Prior to use, 1 ml of enzyme solution was diluted 25 times with phosphate buffer ph 7. Chapter4 antidiabetic activity of the selected plants by. Chapter4 antidiabetic activity of the selected plants by in. The management of blood glucose level is the hallmark in the treatment of this disease. Pdf the aim of the present study was to investigate. Aug 27, 2019 puerh tea was presumed to have antihyperglycemic effects via inhibition on alpha amylase and alpha glucosidase. Inhibition of these enzymes delay and in some cases halt carbohydrate digestion thus prolonging overall carbohydrate digestion time causing a reduction in the rate of glucose absorption and consequently reducing postprandial plasma glucose rise henfeld et al. Nowadays, alpha glucosidase inhibitors like acarbose. Phenolic components have shown effective inhibition of. Samples, standard solutions gamma and alpha mangostin or positive control acarbose with different concentrations 50 l, tables 1 and 3, glucosidase 50 l, 2unitml and phosphate buffer. The results of the present study indicate that, fraction 2 of a.
Colorimetric ab174093 activity assay kit alphaglucosidase. In vitro alpha glucosidase inhibition and freeradical scavenging. Unlike amylase inhibition, the lineweaverburk plots indicated that the peptides reduced glucosidase activity through a non. In vitro studies on alpha amylase and alpha glucosidase. Glucosidase inhibitory activity of polyphenols from the. Alpha glucosidase inhibition assay the spp and ibpp fractions inhibited the alpha glucosidase activity in a dosedependent manner. Antioxidant, amylase and glucosidase inhibitory activities. Alpha glucosidase activity assay the alpha glucosidase dependent oxidation of pnitrophenyl adglucopyranoside was measured to assess alpha glucosidase activity. The most potent antioxidant activity was demonstrated by neptunia oleracea with an ic 50 of 35. There are a large number of natural products with alpha glucosidase inhibitor action. Department of chemistry, university of azad jammu and kashmir, muzaffarabad, department of zoology, university. This is an easy, quick and highthroughput capable kit that can measure 0.
Acarbose works by slowing down the absorption of starchy foods from the intestine. Im currently doing the assay for inhibition of glucosidase. They slow the digestion of carbohydrates and delay glucose absorption. Alpha amylase and alpha glucosidase enzyme inhibition and antioxidant potential of 3oxolupenal and katononic acid isolated from nuxia oppositifolia by ali s. Puerh tea was presumed to have antihyperglycemic effects via inhibition on alphaamylase and alphaglucosidase. Analysis of variance anova, tukeys honestly significant difference hsd test. The inhibition amylase activity was assayed using soluble starch as substrate according to the procedure described previously by siow et al. Alphaglucosidase is one of the enzymes responsible for breaking down carbohydrates to smaller sugar particles like glucose, in order for the carbohydrates to be absorbed. Paper open access related content inhibition of glucosidase. It is usually used when a healthy diet and physical activity alone has been unsuccessful, although it is sometimes used together with a sulphonylurea. Alpha glucosidase inhibition assay theglucosidase inhibitory activity was carried out spectrophotometrically using pnpg as substrate zhou et al. Alphaglucosidase inhibitor definition of alphaglucosidase.
In vitro assay of alphaglucosidase inhibitor activities. Prior to use, 1 ml of enzyme solution was diluted 25. Alphaglucosidase assay of extracts was carried out according to method of bachhawat et al with slight modification. Antidiabetic antioxidant and phytochemical profile of. Phytochemical screening, alphaglucosidase inhibition, antibacterial and antioxidant potential of ajuga bracteosa extracts volume. In alpha glucosidase inhibition assay, each compound demonstrated noteworthy alpha glucosidase inhibitory effect ic 50 4. Schematic flow chart of amylase enzyme inhibition assay procedure 4. Screening of glucosidase inhibitory activity from some. Alphaglucosidase inhibition assay principle and procedure. For example, research has shown the culinary mushroom maitake grifola frondosa has a hypoglycemic effect. Alphaamylase assay kit measurement alphaamylase ceralpha. Alpha glucosidase inhibitors work by competitive and reversible inhibition of these intestinal enzymes. Alphaglucosidase activity assay the alphaglucosidase dependent oxidation of pnitrophenyl adglucopyranoside was measured to assess alphaglucosidase activity. Diabetes mellitus is a metabolic disorder of glucose metabolism.
Alphaglucosidase inhibition in this study, compound 2,2bisbenzothiophne 1 exhibited potent activity with ic50 5 0. Pdf alphaamylase and alphaglucosidase enzyme inhibition. Alphaglucosidase inhibitors work by competitive and reversible inhibition of these intestinal enzymes. Ajuga bracteosa, a medicinal herb, is used by local community to cure a number of diseases such as inflammation, jaundice bronchial asthma, cancer and diabetes. The isolated bioactive compound showed significant. Alphaamylase inhibition potential of the morinda lucida extracts was determined figure 1.
Dglucopyranoside that is hydrolyzed specifically by. Alqahtani 1,2, syed hidayathulla 1, md tabish rehman 2, ali a. In the case of diabetes, alpha glucosidase is a vital enzyme essential for cleavage of maltose to glucose for the absorption into the blood stream in the small intestine. The antidiabetic effects of the seed, flesh, rind and leaf of yellow flesh watermelon extracts were evaluated through the inhibition of alpha amylase. Watermelon citrullus lanatus consumption as shown by different studies is attributed to many health benefits, like in prevention of hypertension, cancer, cardiovascular diseases, and even type 2 diabetes due to its phytochemical constituents. The same procedure was repeated for the gallic acid standard solution 500, 250. Briefly, 100% confluent cells were rinsed 3 times with pbs. The present study indicated that artocarpus heterophyllus could be useful in. Elgamal 2, shaza almassarani 2, valentina razmovskinaumovski 3, mohammed s. All product and stock numbers, unless otherwise indicated, are sigma product and stock numbers. The alphaglucosidase inhibition assay was performed using a method from schmidt, lauridsen, dragsted, nielsen, and staerk 2012a.
In vitro assay of alphaglucosidase inhibitor activities of three seagrasses from banten bay, indonesia. Alphaamylase and alphaglucosidase enzyme inhibition and. Alphaglucosidase inhibitor synonyms, alphaglucosidase inhibitor pronunciation, alphaglucosidase inhibitor translation, english dictionary definition of alphaglucosidase inhibitor. Alpha glucosidase inhibition assay principle and procedure. In abcam s alphaglucosidase activity assay kit colorimetric ab174093. The alphaglucosidase inhibitors are contraindicated in patients with diabetic ketoacidosis, inflammatory bowel disease, ulcerative colitis, intestinal obstruction, any chronic intestinal disease disrupting digestionabsorption, any condition that may deteriorate as a result of. However, no integerated literatures were published to substantiate such presumption. Samples, standard solutions gamma and alphamangostin or positive control acarbose with different concentrations 50 l, tables 1 and 3, glucosidase 50 l, 2unitml and phosphate buffer. The reason maitake lowers blood sugar is because the mushroom naturally contains an alpha glucosidase inhibitor. Alpha glucosidase inhibition assay the alpha glucosidase inhibition assay was performed using a method from schmidt, lauridsen, dragsted, nielsen, and staerk 2012a. Acarbose works by slowing down the absorption of starchy. The ability of berry extracts to inhibit alphaglucosidase. Briefly, rat intestinal acetone powder 500 mg was mixed with 10 ml sodium phosphate buffer ph 6. Alpha glucosidase activity can be measured invitro by determination of the reducing sugar glucose arising from hydrolysis of sucrose by glucosidase enzyme.
Antidiabetic antioxidant and phytochemical profile of yellow. Confluent caco2 cell grown on 24well plates were used for. This results in a smaller and slower rise in blood glucose levels following meals, and effectively throughout the day. The purpose of the present study was to investigate the inhibitory effect of morinda lucida leaf. This extract also showed the highest free radical scavenging. In abcam s alpha glucosidase activity assay kit colorimetric ab174093. Alphaamylase and alphaglucosidase enzyme inhibition and antioxidant potential of 3oxolupenal and katononic acid isolated from nuxia oppositifolia by ali s. The assay procedure shows an excellent correlation with the farrand, the falling number. Alphaglucosidase activity assay kit ab174093 abcam. In this assay, glucosidase activity is determined by a reaction in which glucosidase hydrolyzes pnitrophenyl dglucopyranoside resulting in the formation of a. Alpha glucosidase inhibition assay percent inhibitory activity of. The test compound was added into the assay mixture containing 50 mm phosphate buffer ph 6. The inhibition of amylase by all the extracts at lower concentrations 0.
Phytochemical screening, alphaglucosidase inhibition. Hence, further studies may throw light on the antidiabetic potential of a. Inhibitory effects of puerh tea on alpha glucosidase and. Kokab hafeez, saiqa andleeb, tahseen ghousa, rozina g. Alphaglucosidase inhibition assay the spp and ibpp fractions inhibited the alphaglucosidase activity in a dosedependent manner. Alphaglucosidase activity can be measured invitro by determination of the reducing sugar glucose arising from hydrolysis of sucrose by glucosidase enzyme. These chemicals were previously reported in seedcases of garcinia mangostana and had been reported to display alpha glucosidase inhibitory effect ryu et. The glucosidase activity assay kit provides a simple and direct procedure for measuring glucosidase activity in biological samples. Free, esterified, etherified and insolublebound phenolics from three samples, namely meal from whole peanuts mwp, meal from dryblanched peanuts mdbp and peanut skin ps were evaluated for their total phenolic content tpc, and phenolic profile using hplcdadesimsn. Alpha glucosidase inhibition assay the glucosidase inhibitory activity was carried out spectrophotometrically using pnpg as substrate zhou et al. Alphaglucosidase inhibitory and antioxidant activities of various. Cmpe may exert hypoglycemic effects through the same mechanism. G5003, g66, g7256, g8889, g0660, and g3651 principle.
Alpha glucosidase inhibition assay, antibacterial activity, antioxidant activity, phytochemical screening, ajuga bracteosa. Linear regression based on test solutions of ethanolfree extract on percent inhibition in. Thereby, acarbose as a alpha glucosidase inhibitor can be used in diabetic. Characterization and evaluation of mycosterol secreted. This may be achieved through the use of oral hypoglycemic drugs such as biguanides, insulin secretagogues, and. Overproduction of free radical species can be damaging to host cells and lead to various ailments such as inflammatory, aging and diabetes zhou et al.
The inhibition of glucosidase activity was determined using the modified published method. Among 37 compounds identified 10 were reported for. Total phenolic content, alpha glucosidase inhibitory effect, and freeradical. The percentage inhibition was plotted against the concentrations of each sample to determine the concentration required to inhibit 50% of the. Ghadyale v, takalikar s, haldavnekar v, arvindekar a.
Artocarpus heterophyllus showed the greater % inhibition of the alpha glucosidase enzyme compared to other plant extracts. For the control assay, the inhibitor was replaced by an 1 equal volume of pbs. Alphaamylase and alphaglucosidase enzyme inhibition and antioxidant potential of 3oxolupenal and katononic acid isolated from nuxia oppositifolia. The in vitro bioevaluation against alpha glucosidase showed strong inhibitory activities of 8, 10, and 7, with the half inhibitory concentration ic 50 values of 11.
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