This standard specifies the determination of diethylstilbestrol residues in livestock and poultry meat by high performance liquid chromatography (HPLC). This standard is applicable to the determination of diethylstilbestrol residues in fresh chicken, beef, pork and mutton. The detection limit is 0.25 mg/kg. GB/T 5009.108-2003 Determination of diethylstilbestrol in livestock and poultry meat GB/T5009.108-2003 Standard download decompression password: www.bzxz.net

National Standard of the People's Republic of China GB/T 5009.108-2003 GB/T14931.2-1994 Determination of diaethylstilbestrol in livestock and poultry meat flesh2003-08-11 Issued
Ministry of Health of the People's Republic of China
China National Chemical Industry Association
2004-01-01 Implemented
CB/T5009.108-2003
This standard replaces GB/114931.2-1999 Preparation of high-hexenol content meat 8: Compared with (B/T14931.2-1994), the main modifications of this standard are as follows: The Chinese name of the standard is modified, and the Chinese name of the standard is changed to Determination of hexenol in poultry 3 According to GBT20001.4-200 18 Standard Writing Plan Part 4: Chemical Analysis Method 3 The structure of the original standard was revised.
This standard was jointly proposed by the Ministry of Health of the People's Republic of China. Drafting units of this standard: Liaoning Provincial Food and Drug Administration, Shenyang Municipal Health and Epidemic Prevention Station, Jieshan Municipal Health and Epidemic Prevention Station: This standard was drafted by the following persons: Yongxin, Che Xueban, Feng, and Liu Zhaipao. The original standard was drafted in 2011. This standard was specially designed for the determination of diethylstilbestrol in poultry meat.
This standard specifies the use of high performance liquid chromatography (HPLC) method to determine the diethylstilbestrol residues in poultry wing meat GB/T 5009.108—2003
This standard is used to determine the residual content of ethylenic estradiol in fresh chicken, pork, and mutton: the detection limit is 0.25g/kg. 2. After the selection of the pulp, the whole body is extracted and filtered to obtain the ethylenic estradiol. After external measurement and identification, the photometry is measured at 1°C. Under the same conditions, a working curve is drawn. The ethylenic estradiol content and the absorbance value are positively correlated within a certain concentration range. The sample is compared with the working curve for quantitative analysis. 3. Reading agent
3.1 Alcohol
3.2 0.43μl/L sodium dihydrogen phosphate (NH.PO,-2H,0) is taken and 1K sodium dihydrogen phosphate is dissolved in water at 50°C. 3.3 Deoxygenase
3.4 ​​Deoxysuccinate (DES) standard solution: Weigh 100 mL of DES molten in 10 mL volumetric flask, add methanol to the mark, mix, and add 1.1 mg of DES per liter in ice. 3.5 Deoxysuccinate (IES) standard working solution: Take 10 mL of DES solution, add it to a 10 mL volumetric flask, add methanol to the mark, mix, and add 100 g of DES per mL. 4 Apparatus
4.1 High performance liquid chromatography: Other apparatus. 4.2 Small granulator.
4.3 Small pulverizer. ||t t||4.4 Electric sterilization machine.
4.5 Centrifuge.
5 Analysis steps
5.1 Extraction and purification
Weigh: (±0.1g) minced (small, mm> meat sample. Put into 50ml. Centrifuge with stopper, add 1C.50ml methanol, stir well, remove the supernatant, add 10.00mL methanol, mix well and remove bacteria for 20min, centrifuge at 10001/min for 0min, and remove the supernatant. If there is any confusion at this time, centrifuge again for 10min, pass the supernatant through 0.5mFH filter membrane, and use.
5.2 Chromatographic conditions
5.2.1 External examination: Detection wavelength 230nm. 5.2.2 Flow rate: 0.01AUFS.
5.2.3 Mobile phase: methanol-1.04Srm>t/T. Acetic acid=hydrogen hydride (70+30). Adjust pH to 5 with aldehyde (NaHP0,·2TT:D) aqueous solution, 0.45m filter),
5.2.4 Flow rate: 1 mL/mia.
5.2.5 Injection volume, 20
GB/T 5009.108—2003
5.2.8 Chromatography.tJ.cCDsCu<5m>6.2mmx150mm stainless steel column. 5. 2,7 Temperature: real temperature.
5.3 Drawing of standard curve
Take 500g of chopped meat (5.0g each) and put it into 59ml centrifuge. Add 1,000mL of standard solution with different concentrations (6.0, 12.0, 18.0, 24.0ug/ml) respectively, and do whitening at the same time. The total concentration of methyl alcohol is 20.00mL. Make its concentration to be 0.60.9.1.20g/mL. Extract according to the treatment method. 5.4 Determination
Take 20μL of sample respectively and inject it into HPLC to measure the peak quotient of DES concentration. DES concentration vs. peak height is plotted as curve. At the same time, take 20μL of sample and inject it into HPLC. The peak height obtained from the working curve is R8.23E. 5.5 Calculation
Calculate according to the following formula:
4×1009
1000%100c
X--Diethylstilbestradiol content in the sample, in millimoles (g/g);
Hexadecene content in the sample volume, in nanograms (%);
The total volume of the sample, in grams (g);
Injection volume, in microliters (uL);
The total volume extracted in the sample, in units of kPa (kPa). 5.6 Chromatography
Chromatogram Figure 1.
Entire peak bzxZ.net
Concentrated peak,
Sample standard has been added,
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