Diabetes has been considered to be one of the major risks of cataract and retinopathy. Alpha amylase and alpha glucosidase inhibitory effects of. Antidiabeticactivities of medicinal plants weredetermined byamylase and glucosidase inhibition assay. Alpha glucosidase activity assay the alpha glucosidase dependent oxidation of pnitrophenyl adglucopyranoside was measured to assess alpha glucosidase activity. The rate of formation of glucose is measured in a hexokinaseglucose6phosphate dehydrogenase system. In abcams alphaglucosidase activity assay kit colorimetric ab174093. The inhibition of glucosidase potential shown maximum activity of 90. Glucosidase inhibitory activity of polyphenols from the. The glucosidase activity assay kit provides a simple and direct procedure for measuring glucosidase activity in biological samples.
Antioxidant, amylase and glucosidase inhibitory activities. Dglucopyranoside that is hydrolyzed specifically by. In vitro assay of alphaglucosidase inhibitor activities. As a mixture, the three compounds inhibited to evaluate the structureactivity relationship of the compounds, similar compounds were tested for eugenoa properties using dpph free radical scavenging and beta carotene bleaching protection bc and antidiabetic activity using inhibition of alpha glucosidase activity agthe results are. In vitro studies on alpha amylase and alpha glucosidase. We have developed a new fluorogenic substrate for the alphaglucosidase enzyme assay, resorufin alpha dglucopyranoside. The alpha glucosidase inhibition was assayed using purified enzyme according to the modified method described by matsui et al. Chapter4 antidiabetic activity of the selected plants by. The method is based on the specific recognition between 1,4phenylenediboronic acid pdba and 4aminophenyl. Although acarbose can be used as monotherapy, it is typically added to insulin or other oral agents to achieve glycemic control in type 2 diabetes. Glucosidase hydrolyzes the substrate mix to release the pnitrophenol that can be measured colorimetrically od 410 nm. Paper open access related content inhibition of glucosidase.
For example, research has shown the culinary mushroom maitake grifola frondosa has a hypoglycemic effect. This is an easy, quick and highthroughput capable kit that can measure 0. Inhibition of alphaglucosidase and antioxidant test of stem bark. Active compounds of plants have potency as antidiabetic that can be used in diabetes mellitus treatment.
Nowadays, alpha glucosidase inhibitors like acarbose. Antidiabetic activity and phytochemical screening of extracts. The management of blood glucose level is the hallmark in the treatment of this disease. Diabetes mellitus is a metabolic disorder of glucose metabolism. Emerging strategies for the activity assay and inhibitor.
In abcam s alphaglucosidase activity assay kit colorimetric ab174093. One of the strategies is maintaining postprandial glucose level through inhibition of. Request pdf emerging strategies for the activity assay and inhibitor screening of alpha glucosidase the high incidence of diabetes mellitus has caused widespread concern around the world. The inhibition of glucosidase activity was determined using the modified published method. Such inhibitors which find application in the clinical practice for management of diabetes are known to be associated with various gastrointestinal side effects. Pharm10a6 list the main drug groups used in the treatment. After incubation at 30c for 60 min, the absorbance was then. This inhibitor, known as acarbose, was the result of long screening program for glucosidase inhibitors and has been reported to be quite efficient in controlling blood glucose levels. Glucosidase inhibitory activity of dichloromethane. Colorimetric ab174093 activity assay kit alphaglucosidase. The percentage inhibition of assays showed maximum inhibitory effects 59. Mustafa, anum naseer, irsa shafique and kalsoom akhter affiliation. Can anyone help to find the best protocol for evaluation of alpha.
The reason maitake lowers blood sugar is because the mushroom naturally contains an alpha glucosidase inhibitor. Glucosidase agh from the small intestine of rat was immobilized onto a glutaradehyde ga activated nh 296 well microplate to establish a convenient and rapid agh inhibition assay system. Alpha glucosidase activity can be measured invitro by determination of the reducing sugar glucose arising from hydrolysis of sucrose by glucosidase enzyme. Moreover, the representative strategies for the assay of gaa activity and agi screening are compared. Synthetic and natural product inhibitors of carbohydrate degrading enzymes are able to reduce type 2 diabetes and its complications. One mg of glucosidase saccharomyces cerevisiae, sigmaaldrich, usa was dissolved in 100 ml of phosphate buffer ph 6. The purpose of the present study was to investigate the inhibitory effect of morinda lucida leaf. Phytochemical screening, alphaglucosidase inhibition. A new resorufinbased alphaglucosidase assay for high. Alpha glucosidase inhibitors agis are oral antidiabetic drugs used for diabetes mellitus type 2 that work by preventing the digestion of carbohydrates such as starch and table sugar. Many of the agents used as antidiabetic are either expensive or have side effects. Alphaglucosidase activity assay kit ab174093 abcam. In vitroantidiabetic activity and glycemic index of bee honeys. The percent of inhibition was determined by the following formula.
In the present study, the in vitro study revealed that the extract of g. Bee pollen has potential antioxidative and antidiabetic activity. Feb 22, 2018 alpha glucosidase inhibitors such as acarbose decrease the absorption of carbohydrates from the digestive tract, thereby lowering the aftermeal glucose levels. Sep 23, 2014 a colorimetric method has been established for. 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. Glucosidase inhibition assay was carried out by following the method of elya et al. M and all the experiments were carried out in triplicates. The purpose of the present study was to investigate the inhibitory effect of morinda. The extracts of all parts root, leaf, and stem from three seagrasses were prepared with maceration method using 80% ethanol solvents.
The test compound was added into the assay mixture containing 50 mm phosphate buffer ph 6. The ability of berry extracts to inhibit alphaglucosidase in. Objective of this research is to investigate alpha glucosidase inhibitory activity from syzygium species. Vasicinol 9, which were isolated by assayguided fractionation. The ic 50 values of stem extract was found to be 73. Prior to use, 1 ml of enzyme solution was diluted 25 times with phosphate buffer ph 7. D i a b e tes o f mt l ab n r u o siol journal of diabetes. Glucosidase inhibition assay in an enzymeimmobilized.
Assay principle the change in absorbance is measured at 400 nm according to the following reaction. Alpha amylase enzymatic assay was performed in triplicates method. Screening of glucosidase inhibitory activity from some. We also studied the in vitro antidiabetic activity of s. Emerging strategies for the activity assay and inhibitor screening of. Results revealed that methonolic extract of plants posses inhibitory properties of alpha amylase and alpha glucosidase in vitro. For a long time, potatoes have been portrayed as unhealthy for diabetic patients by.
The highest concentration of 100 mgml tested showed a maximum inhibition of nearly 78. Therefore, it is the need of time to identify and explore the amylase inhibitors from natural sources having fewer side effects. Studies on antidiabetic activity of indian medicinal. Alphaglucosidase and alphaamylase inhibitory activities. Alpha glucosidase inhibition assay principle and procedure. There is only one tablet of this type used, called acarbose. Carbohydrates are normally converted into simple sugars monosaccharides by alpha glucosidase enzymes present on cells lining the intestine, enabling monosaccharides to be absorbed through the intestine. Glucosidase specific inhibitor, the enzymatic activity is greatly reduced which is detected by a decrease of absorbance readings. Below are the links to the authors original submitted files for images.
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. However, the specific components responsible for the hypoglycemic effect still remain unknown. In the colon, bacteria will digest the complex carbohydrates, thereby causing gastrointestinal side effects such as flatulence and diarrhea. Ic50 values for inhibition of alphaglucosidase test are 20. Pdf alpha glucosidase inhibitory activity from syzigium sp. The acetone extract of picralima nitida displayed the most effective inhibition of both. Im currently doing the assay for inhibition of glucosidase. Evaluation of alpha glucosidase inhibitor miglitol for. Evaluation of alpha glucosidase inhibitor miglitol for its efficacy in constipation associated with diabetes. The assay kit provides a rapid, simple and reliable test for highthroughput screening of. Glucosidase inhibitors and aldose reductase inhibitors of. Department of chemistry, university of azad jammu and kashmir, muzaffarabad, department of zoology, university. Add 10 50 l test sample to wells of a 96well plate. Alpha glucosidase inhibitors work by competitive and reversible inhibition of these intestinal enzymes.
The recovered aggregation of aunps could lead to an increasing absorbance at 600 nm and. For the control assay, the inhibitor was replaced by an 1 equal volume of pbs. In this study, we investigated the inhibitory effects of ethanolic extract of the leaves of senna surattensis eess on. The aqueousethanol extract displayed higher inhibition of. The principle of this sensitive assay is based on the reaction of samples with the organic radical dpph 14.
Alpha glucosidase is one of the enzymes responsible for breaking down carbohydrates to smaller sugar particles like glucose, in order for the carbohydrates to be absorbed. Phytochemical screening, alphaglucosidase inhibition, antibacterial and antioxidant potential of ajuga bracteosa extracts volume. All product and stock numbers, unless otherwise indicated, are sigma product and stock numbers. Since alphaglucosidase inhibitors prevent the degradation of complex carbohydrates into glucose, the carbohydrates will remain in the intestine.
There has been a global surge in the number of diabetic cases. It is usually used when a healthy diet and physical activity alone has been unsuccessful, although it is sometimes used together with a sulphonylurea. Acarbose used as the positive control showed ic 50 value of 36. In this assay, glucosidase activity is determined by a reaction in which glucosidase hydrolyzes pnitrophenyl dglucopyranoside resulting in the formation of a. The results of enzymatic assay were expressed as percentage inhibition, which was calculated using the formula. Acarbose works by slowing down the absorption of starchy foods from the intestine. Confluent caco2 cell grown on 24well plates were used for.
The modes of inhibition of both enzymes was determined using lineweaverburke plot. Ic 50 value of acarbose in iaghmaltase assay system, 340430 nm. Recently, the screening of alphaglucosidase inhibitors, from natural products, with excellent. Glucosidase inhibitors isolated from medicinal plants. There are a large number of natural products with alpha glucosidase inhibitor action.
Immobilized agh iagh inhibition study by synthetic inhibitors acarbose and voglibose revealed that the magnitude of inhibition differed from that in the free agh fagh study. Kokab hafeez, saiqa andleeb, tahseen ghousa, rozina g. Agathis dramaga, bogor, indonesia 2biopharmaca research center, bogor agricultural university, jl. This may be achieved through the use of oral hypoglycemic drugs such as biguanides, insulin secretagogues, and. Spleen and liver enzyme activity was lowered in gauchers patients when assayed with the synthetic substrate, 4methylumbelliferal. Alpha amylase inhibitory activitv of some plant extracts. I tried to evaluate an alpha glucosidase assay of the methanol plant extract. In addition, alphaglucosidase is a therapeutic target for type ii diabetes, and alphaglucosidase inhibitors have been used in the clinic as alternative treatments for this disease.
Glucosidase assay buffer by adding 10 l of 100 mm pnitrophenol standard to 90 l of. The results of the present study indicate that, fraction 2 of a. The aim of this study was to examine the inhibition of. Schematic flow chart of amylase enzyme inhibition assay procedure 4. After agh immobilization, remaining ga groups were blocked by. They slow the digestion of carbohydrates and delay glucose absorption. This means that blood glucose levels rise more slowly after meals. Asclepiadaceae has been known to posses potential antidiabetic activity, and the gymnemic acids were reported as the main bioactive components in this plant species.
Researchers are now turning their attention to phytotherapy as a viable alternative in the treatment of hyperglycemia. In vitro inhibitory effects of plantbased foods and their combinations. Alpha glucosidase inhibition works by reducing glucose absorption due to decreased carbohydrate digestion. The alpha amylase inhibition assay showed that the methanolic extracts of cinnamomum zeylanicum. Part of the food science commons recommended citation. Diabetes mellitus is a chronic disease that is becoming a serious global health problem.
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