Some standard content:
[CS 65.120
National Standard of the People's Republic of China
GB/T20196—2006
Determination of saliuomycin in feed
Deteiination of saliuomycin in feed Issued on February 24, 2006
General Administration of Quality Supervision, Inspection and Quarantine of the People's Republic of China Standardization Administration of China
Implementation on July 1, 2006
Standard is used in domestic and foreign countries to determine these bands in feed: GR/T20196—238
The first one is the microbial test method for rat health inspection (explosion method)! The method for testing the salt content of wheat flour, meat and aquatic products exported by the Japanese provincial quality inspection bureau 673 is formulated by the company. The method is based on the principle of microbial detection and the steps of the method were improved. The specifications and conditions of the sample preparation were improved. The technical contents of the improvements are as follows: - The liquid is separated and extracted with alcohol and water. The whole substance is separated and purified by a column chromatography and then cooled. The second method is a high-efficiency chemical method, which was reported by Joura.atch:amangnlyA19 and is similar to the effective chemical method for determining the content of sand in animal feed. It requires typical factors, basic principles, and steps. The method is formulated based on the same method, and improvements are made according to the experimental sample quality. The detailed technology is given below! "This standard specifies the total return of the feed using a combined liquid end, the average limit of each system benefit pre-standard agent, which is larger than the text paragraph: a standard sample and a single extraction station require a teacher to see the white shot requirement A, the above methods are all "vertical": in the standard bomb period B, time (for normative records, network limit A is an informative appendix. This standard is provided by the regional feed chemical technology association, the starting unit of this standard; the national quality supervision and inspection center (Wuyi!. The starting five people of this standard: Liu Xiaomin, Zhang Yong, Gao Li, He·-Sail 1 Pineapple Wei
Determination of salt mold in feed
GR/T 0196—20CG
The standard specifies the method for testing salt content in feed materials using micro-electrochemical methods. Both methods in this standard are applicable to the determination of salt content in mixed feed particles with the same feed additives. The minimum detection limit is 2%. The local quarantine method is used as the reference method. The high-efficiency micro-electrochemical method is also applicable to the determination of salt content in mixed feed particles using micro-electrochemical methods: 2 Normative references
The clauses in the following documents become the clauses of this standard through the use of this standard. For the referenced documents, their use shall apply to the following clauses: Some of the changes (not including the previous certificate) or the proposed version are not used for wood standard liquid. However, the latest amount of these documents must be used in the case of the booklet. The latest version of the standard can be used after the chemical is marked. 4. General rules for food biological testing CB/T5332 Analytical laboratory water preparation and preparation method GT1!1 words, sampling
3 method 1: biological test method! Secondary method: 3.1 principle
le only takes a sample of salt and water, and takes the purified water with air than the original lead, and reduces the large amount of materials in the request! Approval material: method of setting up road Xi (or policy reduction) 3.2 Test phase and materials
Except for the standard test, use one-third of the test and distilled water of the required purity or higher. 3.2.1 Dissolve in water at 27-343F, purify at 0℃ for 1 day, cool to room temperature for 1 month, and then cool to room temperature for 2 months. 3.2.3 Aluminum oxide layer removed in a column
Activated aluminum oxide layer installed in a folding machine 2imm, inner diameter mm, oxidized surface cannot be translated by tapping, high temperature ~m.
3.2.4 Test strains
This heat-resistant bacteria (Barlissierotheinphsiwvu, calitotetit3.2.5 Salt poison standard solution
3.2.5.1 Station two standard independent solution
Contact system standard: 9101263 placed in 33 stock this consultation solution to select the degree, the rate is 1: quantity into the building market, the right expiration, 3.2.2.2 Salt demand standard intermediary
letter transfer center with standard each full (325.! ! 10m. tube bottle, use Bar pregnancy grid to reduce the degree to select its liquid concentration to m. When the limit and this use
CB/I20196-2CC6
3.2.5.3 Salt standard working solution
Note that the intermediate solution required for the transfer order <3.2.5.21, the standard solution (3.2.1) is 5.C℃1gm, 10.mL-3.0g/ml20.0/25./r! Teaching, of which 5.0gm is the standard working wave of the reference solution. The above culture solution must be prepared and used as above: 3.2.6 culture medium
can be prepared at the specified time.
3.2.7 Suspension
Thermolipid therapy: inoculate in flow medium T, culture for (17-1)h at 3 000 t:mn for 15 min, add salt solution, counter-select, and prepare suspension at the optimum temperature. Store in ice. The suspension can be quickly used.
3.2. Preparation of test plates
Before preparing the plates, a standard suspension of 0.101r1 in water was used as the test sample: 3.2. 3 The amount of suspension was prepared by pre-testing, and different amounts of suspension were added to the suspension in the culture refrigeration tank. The culture was fully combined, and after (5 1) hours, the concentration of nuclear particles was increased by 12:10, and the complete characteristics of the separation were described. The blood was cooled and unloaded at 20 °C, and the culture was kept in water until it was coagulated. Then the best culture was added, and the level of the culture was promoted to make it coagulate. The plate can be prepared immediately.
3.3 Instruments and equipment
3.3.1 Sterilization pot or fire box
3.3.2 Wet culture pin: 61=, 5. Keep water?.3.3.3 Rotating point space variable type.
3.3.4 Reciprocating type
3.3.5 Separator J70g (9/mi).
3.3.6 Standard caliper; measuring range 0 mm~2 cr.1: temperature 0.02 ms or beat tooth measurement change, 3.3.7 Maintenance: 90rr.m. bottom production of the whole light elimination effect and then. Total pollution tile benefit. 3.3.8 Noon and other supplements: South diameter 6.m high: m3 small pin tube with outer diameter 7.8mm, 3.3.9 Adjustable volume displacement, 20..1uurl.. method: The base material device can be set up by pulling
3.3.10 Safety inspection of the blood wave potential device,
3.4 Sample preparation
According to 11/1-99.1 actual material sampling method, select the sample weight of the European River method, at least 50 weight at least 10, through the Dm lead span. Along the way, put it into a closed container. Light low alliance protection standby 3.5 Analysis steps
3.5.1 Test concentration preparation
9.5.1.1 Micro-crop actual collection military signature detection as a total of the total input (H4 attachment, implementation 3.5.1.2 Accurate system sampling, anti-accurate to 0.000 [5: "250 ml.H rate two angles, accurately add 20, m.. sample liquid according to 3.? ._!, reciprocate the disassembly device. Multiple ends for 1h. All tablets can be transferred to the prepared chemical reaction tube (3.3): use the column sample to extract iridium %211, select the appearance, use 10, and then press the scale. 3.5.1. 3. According to the concentration (calculated according to the analysis instructions, weigh the sample volume and the original liquid volume, and take a certain amount of the sample. 5. Three and in the fear of the natural release, reduce it to 1. Add an appropriate amount of extraction medium (1.1 weigh: the instrument can be used as required, 2
select the test volume
3.5.2 Preparation of standard curve
CB/T20196—2006
Use 0R1 to change the internal standard silicon working concentration to the special concentration, take a calibration plate for each of the three standard curves as a group, place 6 half cups on each half plate, read the flushing cup on a 2% circular surface with a diameter of 1\normal: 3 of the cups are added with more consideration of the concentration, and the other is eliminated.The standard working solution is called the reference standard solution. There are two working solutions with the same concentration of 3. The standard working solution with the same concentration is used for two calibration levels. Cover the solution with a good seal and incubate it at 4 °C for 1h-2h. Then, incubate it at <55 °C for 17 ± 1h. Remove the pure water and accurately measure the inhibitory effect of the produced soil on the surface of the soil: accurately measure the inhibitory effect of the soil to 0.11m). Calculate the reading of 5.1/m1 on each group of 3 calibration levels (B) and the average of the inhibitory effect of the other concentrations (V), and then calculate the expected concentration (5/m1). The value of all 36 spots (the average value of the expected diameter) is used! The difference between the average diameter (B) of each group of reference bacteria is used to calibrate the average of the readings of the standard working solution (A). The calibration solution A is converted into the calibration coordinates by the limit concentration, and the calibration curve is calculated by the formula 1:
AB--A
The average diameter reading of the bacteria at the point of the calibration is the total average value of the number of single-point calibration curves. The unit is meter (m): The average reading of the standard control group in the calibration wave group is meter (mm): The half-diameter reading of the A
recovery rate liquid is measured in meters (mm): 3.5.3 Determine the
standard. After two tests, place a clean cup on each plate, so that the spacing between the cups is 1 or 5 meters. Fill 3 of them with .1mL sample and add 0.1m. Store the sample on the reference sample with a concentration of 500/m2 and a reference standard 1% wave. Cover the broken tile cover and place it at 4 for 12 hours. Except for the too large parts, measure the diameter of the antibacterial plaque produced by the material (measured accurately, 1m, and corrected. Calculate its average value:
3.6 Method Calculation
3.6.1 If the efficiency environment is reported, the point diameter of the tooth diagram is less than 12mm, that is, the sample shows an inhibitory blue and blue color. The value is greater than or equal to 12: after correction: the sound from the standard mountain cable or the corresponding point of the meter is measured, and the salt content in the final test is calculated according to formula 2). If the result is negative, the average value of the diameter of the point is greater than 12m. It must be corrected and confirmed (the biological product can be used to request the mineral substance of the state outside the state): 3.6.2 The content of the sample in the formula (2): weXn
In the formula:
Sample benefit season system standard : mR/kR: The density of the sample obtained from the standard curve is expressed in kR (/): 7 multiples!
Weigh the sample, in grams!
3.6.3. Calculation of half-values: Mathematical results. 3.7 Repeatability
The absolute value of the two independent test results under the reproducible conditions is the absolute value of the test, (2)
GH/T2C196—2006||t t||The content of salt element is 4.0010"mg/kg, which is the arithmetic value; when the content of salt is >1.Y1mg/kg, it shall not exceed 10% of the arithmetic value. 4 Method 2: High performance liquid chromatography pre-column derivatization test method 4.1 Principle
Use medium and small-scale pre-column derivatization method to carry out pre-column derivatization test with 2 NP as the application point to determine the salt content in the sample with an external detector. 4.2 Reagents Materials and methods Unless otherwise stated, list the following in detail: 4.2.1 Water
Ge/16682
d. 2.2 Ethylene glycol solution
4.2.3 Methanol
Chromatographic purity:
4.2.4 Derivatization test
c=.51g/L: Weigh 5mg 2,4-nitropropene and place in 100ml volume. Dissolve thoroughly in methanol to about 50% elution.
4.2.5 Extraction solution
Use the extract solution and
4.2.6 Salinophil standard solution
4.2.6. Salt standard preparation pool
Accurately weigh each standard sample (content 41016, bone 3) and dissolve in the water until the solution is dissolved, and the solution is stored in the box for validity period of 1/2. 4.2.6.2 Salt standard working solution
Accurately transfer 1.0w:1.-2.00mL of standard preparation solution: 4..S.1>=111ml1. Dilute to the mark with methanol in a bottle, and the concentrations are 10110.m respectively. Prepare and use immediately. 4.2.7 Mobile phase
Transfer 2 mL of ice (high purity) into 1 mL of water and mix with acetyl alcohol at a ratio of 10:90. Degas the mobile phase or perform other degassing treatment.
4.3 Receivers and equipment
4.3.1 Commonly used laboratory spheres
4.3.2 Fresh, female.
4.3.3 Transfer the mobile phase to the test chamber. Www.bzxZ.net
4.3.4 Centrifuge at 7g. /min
4.3.5 Micro-analyzer
43.6 Adjust the volume 105.100--203--0002 Note 3: The two-sensor liquid can be calibrated
3. Liquid scanning: Equipped with four-sensor, source, UV (V) or two-channel tube array detector, S chromatograph particle size 4: Length 5-5 m, diameter 1.5 m.
4.4 Preparation of samples
.1 The samples are collected from the feed and ground by at least 0.233F code, and then evenly packed in a sealed container, protected from light and stored at low temperature. 4.5 Analysis steps
4.5. Extraction
GU/T 20196—2006
4.5.1 Weigh the table at a certain point and record the time accurately to 1, and spray 20.0m sample liquid in the right supply area 4.2. Fill the support and record the liquid in the center of the swing for a while, with 000g (2 533 r/min:4.2 10 mir..1.5..?If the test solution is concentrated, according to the estimated amount of salt contamination according to the labeled extraction filter (1.5.1.1), take the determined amount of sample according to the attached sample volume and extraction energy, transfer to the heat apparatus, and then take the concentrated sample (4.2). Make the test solution contain 10% sodium chloride (4.3)/ml. The derivatization solution can be verified by microfiltration at high temperature for 10) min, then add 200I-nitrogen-based derivatization solution (4.2.4) to the sample source, place it in a 30ml centrifuge at 2℃, take it out and wait for cooling for detection.
If the mixture is found to be more chaotic, centrifuge at 300g (sc331/min) and collect the supernatant for resistance testing. 4.5.3 Standard derivatization
Use an adjustable micro-volume adder (3) to accurately draw 10:23000n of each element standard solution (4.2.5.2 ten ports of the high center, medium and late seed rate 7027 set 1-1 empty wide tube 70l. Lang as a control, amine 4.3.2 period of biochemical method of cattle. 4.5.2 Determination
4.5..1 High performance liquid chromatograph (HIPLL) set up a number of no fixed column: 011sC: pull. Calibrate pressure 4m11: close 151rm internal price 3.51r or it can be beautiful separation grid. Shake leakage: C.
Flow scan: 4.2.71.0l.
Grid detector, ultraviolet (UV) take a tube wire grid detector. Grid measurement wavelength: 3921.
Reverse sample volume. 13 ... -23 ...
1.5,4.2 Qualitative and quantitative determination
4.5.2 I have to take the appropriate biochemical test using strain. 5. Derivatives are made under the standard 1: to ensure that the time-based derivatives are UV-light-specific. Single-point or image point calibration is performed based on the colorimetric area score. 2.6 Result calculation
4. 6.1 The content of the intermediate material is calculated according to the formula (3): F: XVNAV&A
T.xmxyXVi
, the unit is mg/kg, the standard is 1, the unit is gram/kg, mR: kgP
standard industrial production area:
standard industrial production area!, average light model liter (sample area value:
sample derived waste sample description, the median is a small open α); the unit is r:!;
customer and line test volume 4..2 The unit is the nominal concentration multiple
the quality of the sample, the unit is gram). )
GRT20196—2006
4.6.2.1 The results are shown in the table with the arithmetic mean value and retain 3 significant figures. Absolute values of the results of two independent tests under the condition of gravity: when the content of each substance is 4.0% × / kg, it shall not exceed 15% of the arithmetic mean value; when the content of each substance is 4.0% × / kg, it shall not exceed 10% of the arithmetic mean value. (Data record) Sample quantity of test sample: BT2015E-2006 The labeled quantity, sample weight and salt content of the sample are listed in Table 1. Example of dilution of feed. See Figure 4.1, Table 4.2, Table A, feed sample plate, sample weight and dilution of chloranil extract (microbiological test method) Quantity/
Feed category
Pre-combined feed
Formulated feed
Compounded sample
Feed state
Ink agent
Pre-combined feed
Concentrated feed
CaRkgi
7 635 000
a:000
s/tng/:L)
Or technical wine color
Feed sample labeled weight sample weight and chloranil standard extract concentration (IHPLC) station effective center quantity!
Cug/kg?
Ix: non d
: x:3 ano
30: 000
Pain sample
Hot liquid V
Extraction
Associated liquid initial column
Cultivation
Estimated
Micro/
(rgim_)
GB/T20196—2C06
R.1 Culture medium 1
B. 1. 1 Ingredients
Sodium bicarbonate
Citrate
H, 1. 2 Preparation method
Appendix B
! Normative appendix!
Cultivate the culture medium and test phase
- 300 m
Put 7.1% of the above ingredients in a dry test tube, inactivate 13 m, use a standard slant egg,
culture star II
E.2. or
read the egg point
glucose
B.2. 2 Preparation method
I occ rf.
Add a little of the three ingredients in water. Adjust the price to 0.1% H2O and place in a flask. In! 2 It is: 5mm, H.3 Rational saline
Weigh 8.5g fluoride, drop it into 1L water, and heat it at 21℃ for 15min: 6
, 1 Principle
Appendix
[Normative Appendix)
Microbiological autoradiography: Use standard film as control, R value, to determine whether the antibacterial substances present in the saline are indeed salt.
C.2 Conditions of biological autoradiography
C.2.1 Thin layer plate
Stock Q/1125?-e Activation before use h
C.2.2 Spotting amount
L. point.
c.2.3 expand
in the local area
heart 2.4 risk
format a sample wave Sheng standard industry solution phase point East piece sea layer plate first put each piece of thin layer into a product 2cm clear basic energy: then the store this test on the other point 2) m detailed excitement 2.0 "Rml benefit and then the standard format liquid, the standard liquid was point distance =.: m, in the book lungs for expansion T, the construction agent elimination distance nuclear pre Belt 1, take the source layer, pull the layer into the culture place under high temperature and cool it to 50°C under wind-free conditions, then use 1\ to culture the culture medium until it fills the entire layer, keep it flat, and solidify it (2): culturing (7): After culturing, inject salt into the layer and produce the separated species, the position is between the states, and the name is suitable for the protection of the self-developed product from the west.
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