Alpha-amylase helps in the breakdown of carbohydrates to sugar. It is a major secretion of the pancreas and salivary glands. The secretion then enters the duodenum. But, if there is any kind of injury to the pancreas then, the secreted enzyme enters the bloodstream. The increased level of the enzyme may indicate the problem of acute pancreatitis, penetrating peptic ulcers, etc. Similarly, a decrease in the enzyme may indicate the problem of chronic pancreatic disorders.
Everything that I state below will be on the basis of a semi-auto biochemistry analyzer and can differ from one manufacturer to another.
Alpha-amylase acts as a catalyst during the hydrolysis of 2 -chloro -4 nitrophenol salt to chloro nitrophenol(CNP). The intensity of the color of the formed CNP is directly proportional to the activity of the alpha-amylase enzyme.
Normally, you can store the reagents at 2-80C till the expiration date. But, once you open the bottle, the lifespan will reduce to about 28 days (at 2-80C ), including the working solution.
- Wear an apron and surgical gloves before carrying out the measurement.
- Look for the expiry date of the reagents during purchase and measurement time. Suppliers tend to give you reagent kits with a low expiry interval.
- Bring the reagents and samples to room temperature before you can carry out any measurement.
- Always store reagents in the refrigerator when not in use.
If you have two reagents, then you can make a working solution out of it. Dilute enzyme (R2) with the corresponding buffer solution (R1). If you have a single reagent then, you can use it directly.
Add 20 μl of serum/plasma or 8μl of urine to 1000 μl of the prepared working solution (or a single reagent). This composition may slightly differ from one manufacturer to another. After that, feed the solution into a semi-auto biochemistry analyzer and get the result.
If the activity of alpha-amylase is very high (out of range), you need to dilute it with 0.9% NaCl or distilled water in a ratio of 1: y. Then when you get the final result, multiply the value with a dilution factor (1+y).
You will get, an increasing curve in the biochemistry analyzer.