Microdetermination method for phytase in feedstuff

Abstract

The invention discloses a micro-determination method of phytase activity added in feedstuff. The method comprises the following steps: firstly, performing dialysis treatment on a sample by taking acetic buffer solution I as an outer dialysate, reducing inorganic phosphorus content in the feedstuff to eliminate influence of the inorganic phosphorus on the phytase hydrolytic reaction in the feedstuff; secondly, fetching 1ml of liquid to react to replace 0.2 ml of the liquid in the existing method, adding 1 ml of the acetic buffer solution I to replace 1.8 ml of the acetic buffer solution I in the existing method so that blank extinction of the sample is in a reasonable determination range to avoid determination error which is caused by high content of the inorganic phosphorus in the sample blank; and thirdly, adding the known enzyme to the feedstuff, checking whether the known enzyme is lost during the processes of sampling, dialysis and determination by determining the known enzyme. The method can accurately determine the content of the phytase added to the feedstuff.

Description

[technical field] [0001] The invention relates to a micro-measurement method of phytase added in feed. [Background technique] [0002] 1. Application of phytase in feed and its determination method [0003] In recent years, with the development of livestock and poultry breeding and the consumption of phosphorus resources, the price of calcium hydrogen phosphate has been rising year after year, and has become another major feed raw material after protein and energy; on the other hand, the addition of a large amount of inorganic phosphorus in feed has also intensified environmental pollution. Therefore, using microbial phytase to reduce the amount of calcium hydrogen phosphate in the diet is more and more popular. [0004] At present, there are many types and formulations of phytase products. It is particularly important to determine the scientific phytase and the quality inspection and evaluation methods after adding it to the feed to select high-quality and low-cost phytas...

AI Technical Summary

Problems solved by technology

The detection method of phytase derived from Aspergillus was officially released in 2002 (GB / T18634-2002), but because the action site and optimal pH of phytase from different sources are different, GB / T18634-2002 was used to determine the source The phytase in Escherichia coli has a large coefficient of variation in the test results and cannot be accurately determined. Therefore, a fast, simple, highly specific, sensitive, and accurate method is needed to measure the activity of phytase and add it in feed. The activity of phytase in it will be more conducive to the scientific and reasonable formulation and promote the development of phytase industry

[0006] 2. Problems with existing technology for measuring phytase samples in compound (full price) feed

Therefore, the first problem of enzyme activity detection in enzyme feed: the interference of existing impurities (background)

[0012] 2. Problem 2 of Enzyme Activity Determination in Enzyme Feed

This is the second problem of enzyme activity determination in enzyme feed: the problem of too low activity

[0017] 3. Determination of phytase samples in compound (full price) feed with current prior art

However, the composition of compound feed, concentrated feed and additive premixed feed is obviously more complex, and its applicability has been questioned

Using this method to measure "phytase activity added in feed", the result has a large relative deviation and a large coefficient of variation

Images