GB/T 23874-2009Determination of xylanase activity in feed additives - Spectrophotometric method (English PDF)
饲料添加剂木聚糖酶活力的测定 分光光度法
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Issued by
SAMR; SAC
Level / Type
National · Recommended
Issue date
May 26, 2009
Implementation date
October 1, 2009
Scope
GB/T 23874-2009 is the English-translated version of 饲料添加剂木聚糖酶活力的测定 分光光度法.
GB/T 23874-2009 specifies the determination of xylanase activity in feed additives by spectrophotometry. Clause 1 applies it to xylanase feed additive products and gives a minimum detectable amount of 10.0 U/g.
Clause 2 cites GB/T 6682.
The principle in clause 3 is that xylanase degrades xylan into oligosaccharides and monosaccharides, that the reducing sugars formed give a colour reaction with 3,5-dinitrosalicylic acid (DNS) reagent in a boiling water bath, and that the depth of colour is proportional to the reducing sugar produced and so to the xylanase activity, which is therefore obtained by measuring the colour intensity of the reaction solution spectrophotometrically; the same clause defines one unit of enzyme activity, U, as the amount of enzyme that releases 1 micromole of reducing sugar per minute from a xylan solution of 5 mg/mL at 37 degrees C and pH 5.50.
Clause 4, reagents and solutions, requires analytical grade reagents and water meeting the grade two requirements of GB/T 6682, then sets out the preparations: a sodium hydroxide solution from 20.0 g made up to 100 mL, an acetic acid solution from 0.60 mL of glacial acetic acid made up to 100 mL, a sodium acetate solution from 1.36 g of sodium acetate trihydrate made up to 100 mL, an acetic acid and sodium acetate buffer made up to 2000 mL and checked and adjusted to pH 5.50 with those two solutions, a xylose solution from 1.000 g of anhydrous xylose dissolved in that buffer, and a xylan substrate from 1.00 g of xylan with 0.32 g of sodium hydroxide and 90 mL of water under magnetic stirring.
A footnote to the xylan adds that the trade name given is for the convenience of users, does not imply endorsement of the product, and that equivalent products having the same effect may be used.
The draft breaks off in the middle of that last preparation, so the clauses after clause 4 are not shown, and the foreword records only that Appendix A is informative and that the standard falls under the national standardization technical committee for the feed industry. It was issued on 2009-05-26 and has applied since 2009-10-01.
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Document preview — GB/T 23874-2009
National Standard of the People's Republic of China
- ICS
- 65.120
- Classification
- B 46
Issued by: State Administration for Market Regulation; Standardization Administration of the PRC
Contents
- Foreword
- 1 Scope
- 2 Normative references
- 3 Principle
- 4 Reagents and solutions
Foreword
Appendix A of this standard is an informative annex.
This standard by the National Standardization Technical Committee Feed Industry and focal points.
1 Scope
This standard specifies the viability assay by spectrophotometry feed additive xylanase.
This standard applies to xylanase feed additive products, the minimum detectable amount of 10.0U/g.
2 Normative references
The following documents contain provisions which, through reference in this standard and become the standard terms. For dated references, subsequent Amendments (not including errata content) or revisions do not apply to this standard, however, encourage the parties to the agreement are based on research Whether the latest versions of these documents. For undated reference documents, the latest versions apply to this standard.
Laboratory use specifications and test methods GB/T 6682 Analysis
3 Principle
Xylanase can xylan degrading into oligosaccharides and monosaccharides. Reducing oligosaccharides and monosaccharides in a boiling water bath conditions with 3,5-dinitro Salicylic acid (DNS) reagent color reaction. The amount of reducing sugars and the reaction liquid color depth enzyme produced is proportional to the generation of reducing sugars Volume and vitality and the reaction mixture is proportional to xylanase. Thus, the color intensity of the reaction solution by colorimetric spectroscopic measurement, the reaction solution can be calculated Xylanase vitality.
At 37 °C, pH 5.50 under the conditions of xylan per minute from a solution at a concentration of 5mg/mL reduction in degradation release 1µmol Sugar amount of enzyme required for the enzyme activity unit U.
4 Reagents and solutions
Unless otherwise specified, the reagents used were of analytical grade water are in line with two water GB/T 6682 stipulated.
Weigh sodium hydroxide 20.0g. Dissolved in water, dilute to 100mL.
Draw glacial acetic acid 0.60mL. Dissolved in water, dilute to 100mL.
Weigh sodium acetate trihydrate 1.36g. Dissolved in water, dilute to 100mL.
Weigh sodium acetate trihydrate 23.14g, glacial acetic acid was added 1.70mL. Add water to dissolve, dilute to 2000mL. PH of the solution was measured.
If the deviation from pH 5.50, and then a solution of acetic acid (4.2) or sodium acetate (4.3) was adjusted to 5.50.
Weigh anhydrous xylose 1.000g, plus acetic acid - sodium acetate buffer solution (4.4) to dissolve the volume to 100mL.
1) SigmaX0627 is a trade name, this information is given for the convenience of users of this standard does not imply endorsement of the product. If other products Can have the same effect, you can use these equivalent products.
Weigh xylan (SigmaX06271)) 1.00g, was added 0.32g of sodium hydroxide, then 90mL of water was added, with magnetic stirring while slowly
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This preview omits tables, figures, formulas and parts of the technical clauses. The complete document — 15 pages — is available in the English PDF.
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