Custom Search

Sunday, November 22, 2009

How to estimate beta -Amylase

B-amylase from higher plants catalyses the hydrolysis of 1, 4 – glycosides bands to maltose. It is an examples i.e. splits maltose units from non-reducing end. When a branch point is reached, the hydrolysis ceases giving limit dertin. The name B-amylase is given because the initial product of hydrolysis is B-maltose which lutes on comes in equilibrium with maltose and forms an equilibrium mixture. The activity of enzyme is determined by estimating the reducing sugar produced. The amylase activity is expressed in terms of mg. of lactose liberated in 3 mins. By 1 ml of enzyme solution at optimum PH and temperature.

Reagent: - 1% starch solution [substrate], 5% homo genet of moony [enzyme]. DNS reagent, Buffer PH 4.8, Maltase Standard solution [1mg / ml].

Procedure: - Standard curve.
Prepare a series of test tubes with varying conc. [0 – 2 mg] of maltose. Make up volume to 3 ml. Add of DNS reagent and heat on coater bath for 10 mins. Add 5 ml water to all tubes and determine absorbance. Plot the graph of absorbance against concentration of maltose.

Assay of B-Amylase: - Incubate 1 ml of enzyme solution with 0.5ml of starch and 0.5 ml buffer stop the reaction at the end of 3 mins. By adding 2 ml DNS reagent. Heat the solution in a boiling water bath for 10 mins. Cool and read absorbance (after proper dilution) against blanks.
5% extract of germinating moog.
Effect of varies Ions on Amylase Activity study of effect of Mg++, Zn++, Pb++ and Cl icons on amylase activity.

Many enzymes are stimulated by the presence of simple molecules, ions. These are enzyme stimulants i.e. cultivators. Many compounds have the ability to combine with certain. Enzyme and then block catalysis by that enzyme. Such comps are called inhibitors.

Reagent: - 1) Enzyme (5% homogenate of moog) 2) Buffer of PH 4.8
3) 0.5 % Nacl 4) H.starch (substrate)
5) 4 % znso4 6) 2 % Pb (No3)2
7) 4 % Mg So4

Procedure: - Prepare series of tubes as follows add 1 ml of enzyme extract to all tubes except blank. Incubate all tubes at 370 c for 20 mins. Add 1 ml of DNS reagent and 1 ml of Nacl to all tubes. Add 1 ml of enzyme extract to blank. Heat all tubes on a boiling water bath for 10 mins. Read absorbance at 540 nm.

Result: - The effect of ions on B-amylase activity from moong is
Activators are Cl - & Mg ++
Inhibitors are Pb ++ & Zn++


This website is a guide for pharmaceutical manufacturing and for pharmacy students and pharmaceutical regulatory affairs proffesionals and for every one associated with pharmaceutical industry , we recommend you to subscribe to this website by entering your email ID in the box provided so that you will receive our articles as and when our website is udated.

This website is most favorite Amongst proffesionals in almost all pharmaceutical companies in USA , Pharmaceutical companies in UK , Pharmaceutical companies in Japan , Canada , UAE , Pharmaceutical companies in India 

Following are some of articles which;will be useful for you in further understanding of aspects of sterile dosage form manufacturing

Pharmaceutical Validation

Cleen Room Classification

Pharmaceutical Industry

Pharmaceutical Manufacturing

Clinical Trials

Media fill

Regulatory Affairs in Pharmaceutical Industry

No comments:

Posted by: Martin

How to sucessfuly activate and receive our articles on pharmaceuticals by E-Mail

Dear readers if you wish to receive our articles on pharmaceutical automatically into your email Inbox , as and when we write a new article over this website, then follow these 3 simple steps.

1.You should write your Email ID in the box provided and push the subscribe butten

2.Enter the text code that we show you in a small new browser window after you enter your email ID.

3.Log in to your email and Cheke our verification email sent by "FEEDBURNER" , there you should verify the link we send you by email.

Enter your email address get our articles by email whenever this website is updated

Delivered by FeedBurner

Join the list of our readers from Universities ,Research and Development centers Pharmaceutical Companies from all over the world.

Pharma Research Regulatory Guidelines Website
Texas Dallas, TX 75211 US
Email: Website:
Copyright © 2008-2015 all rights reserve