This standard specifies the method for determining bacillus titanium zinc in feed by liquid chromatography. This standard is applicable to the determination of bacillus titanium zinc content in pig and chicken additive premixes. NY/T 726-2003 Determination of bacitracin zinc in feed by high performance liquid chromatography NY/T726-2003 Standard download decompression password: www.bzxz.net
This standard specifies the method for determining bacillus titanium zinc in feed by liquid chromatography. This standard is applicable to the determination of bacillus titanium zinc content in pig and chicken additive premixes.

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Agricultural Industry Standard of the People's Republic of China
NY/T 726—2003
Determination of bacitracin zinc in feeds--High performance liquid chromatography
Determination of bacitracin zinc in feeds--Iigh performance liquid chromatography2003-12-01Published
Ministry of Agriculture of the People's Republic of China
2004-C3-1Implemented
NY/T 726—2:: 3
This standard refers to AOAC65, 1168 (1982 "Determination of bacitracin zinc in premixes" and related domestic and foreign documents, and is formulated based on the actual situation in my country.
Appendix A of this standard is a normative appendix.
This standard is proposed by the Ministry of Agriculture of the People's Republic of China. The National Feed Industry Standardization Technical Committee is responsible for the preparation of this standard by the National Feed Quality Supervision and Inspection Center (Beijing). The main drafters of this standard are: Gao Sheng, Rao Zhihua, Fan Mai, Li Libei, Yang Shuming. 1 Scope
Determination of bacitracin zinc in feed
High performance liquid chromatographywwW.bzxz.Net
This standard specifies the method for determining bacitracin zinc in feed by liquid chromatography: the standard is applicable to the determination of bacitracin zinc in pig and chicken additive premixes. 2 Normative references
NY/T 726-2$c3
The following documents are used as standard clauses by non-standard reference. The referenced documents are dated, and all subsequent amendments or revisions are applicable to this standard. However, all parties who have reached an agreement on this standard should check whether the latest versions of these documents can be used. For any undated referenced documents, the latest versions are not applicable to this standard (: B/T6E82 Specification for Analytical Experimental Water Test Method Gt.F11699.1 Material Sampling Method
3 Principle of the Method
The natural zinc in the additive pre-excreted material can be extracted with an organic solvent system, centrifuged, and the supernatant is used for liquid chromatography. 4 Reagents and Materials
The following reagents are all analytical. Water is required for the following (/secondary water, 4.1 Phosphorus acid concentrate: H5. Weigh 1.59 g dihydrogen phosphate and x.5 g potassium dihydrogen phosphate. . Dissolve 4.2% phosphate in 11% water EITA buffer: DI1.3. Weigh 13.5% potassium phosphate and 3.5% sodium ethylenediaminetetraacetate and dissolve in ice. 4.3 Bacillus swelling standard solution: 4.3.1 Preparation of standard solution: 150.0 mg of non-precise zinc standard. Dissolve with extraction solution and make up to 190%... The concentration of zinc in the solution is 300%. 4.3.2 Stimulation: aspirate 10 ml of the standard stock solution (41.3.1) and make up to 10 ml with the extraction liquid. The concentration of zinc in this solution is 2:300 mg/L.
4.3.31 Preparation liquid: aspirate 3.20 ml of the intermediate solution (4.3.2), 1.mml and 5.0 ml with the extraction liquid. Make up to 20 ml. The final concentrations of the solution are 1 mg/L, 3 mg/L and 1.5 mg/L respectively. 4.4 Solvent system: prepare according to the table. Each product shall be marked with a solvent of 100 ml. The volume shall be used in a small amount: -1. Table 1 Solvent system 113.1 Buffer solution Glass gel A Deep phosphate A Liquid B Hot water NY/T 726-2C03 Instruments and equipment Analytical balance: sensitivity 0.001; sensitivity 0.0001, 5.2 Ultrasonic water bath 5.3 Centrifuge: 4000 r/min
5.4 High performance liquid chromatograph: equipped with UV detector and 2.5×150mmCm chromatographic sample, 6
Sample preparation
According to B/T, 499.1 Take a representative feed sample, crush it and pass it through a C.45m pore size sieve, mix it thoroughly and put it into a ground-mouth bottle.
7 Analysis steps
7.1 Extraction
Weigh 2.000)g of the sample, accurately add 10mL of the extraction solution, and shake it thoroughly to mix the sample and the extraction solution completely: place it in an ultrasonic water bath and extract it for 15 min, shaking it from time to time, take the upper liquid and centrifuge it at 4000 r/mit. Centrifuge it for 5 min, and directly take it for high-performance liquid chromatography determination.
7.2 Determination
7.2.1 Liquid chromatography conditions
a) Chromatographic column: 2.5x150 mm, 1 μm cm chromatographic column. Mobile phase: Solvent system Table 1A solution: 1-59 solution was mixed. The pH was adjusted to .8 with sodium hydroxide solution (NaOH) = 11 mol/l).
e) Detector: Adjustable wavelength detector, detection wavelength is 254 nm. d) Feed volume: 20 l.
7.2.2 Qualitative and quantitative methods
Under the above conditions, the peak retention time of the two main active components of bacitracin zinc (see Appendix A) was compared for qualitative confirmation. The peak area sum of the main active components was compared with the peak area sum of the peak of the bacitracin zinc standard with similar concentration for quantitative calculation. 8. Expression of analytical results The content of bacitracin zinc in the sample expressed as mass fraction is calculated according to formula (1): A..V
Wherein:
-content of bacitracin zinc in the sample expressed as mass fraction, in milligram per kilogram (mg/kg); A-additive value of bacitracin zinc in the sample;
A,-additive value of standard bacitracin zinc;
m-injection mass of standard bacitracin zinc, in nanogram (ng); V-volume injected into HIFLC, in microliter (μL); n-mass of the weighed sample, in gram (g); V
fixed volume of the sample, in milliliter (ml). 9. Allowable difference
relative deviation of the results of two parallel tests. 2
Appendix A
(Normative Appendix)
Standard Spectrum of Bacitracin Zinc
Standard Spectrum of Bacitracin Zinc
NY/T 728-23-3
NY/T726-2003
People's Republic of China
Produced Standard
Determination of Bacitracin Zinc in Feed
High Performance Liquid Chromatography
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