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GB 29689-2013 English PDF

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GB 29689-2013: Determination of Thiamphenicol residues in milk by High Performance Liquid Chromatographic method
Status: Valid

Standard ID	Contents [version]	USD	STEP2	[PDF] delivered in	Standard Title (Description)	Status	PDF
GB 29689-2013	English	209	Add to Cart	3 days [Need to translate]	Determination of Thiamphenicol residues in milk by High Performance Liquid Chromatographic method	Valid	GB 29689-2013

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Basic data

Standard ID	GB 29689-2013 (GB29689-2013)
Description (Translated English)	Determination of Thiamphenicol residues in milk by High Performance Liquid Chromatographic method
Sector / Industry	National Standard
Classification of Chinese Standard	C53
Classification of International Standard	67.020
Word Count Estimation	9,921
Quoted Standard	GB/T 6682; GB/T 1.1-2000
Adopted Standard	GB/T 6682; GB/T 1.1-2000
Regulation (derived from)	China Food & Drug Administration [2013] No. 234, November, 1, 2013
Issuing agency(ies)	Ministry of Agriculture of the People's Republic of China, National Health and Family Planning Commission of the People's Republic of China
Summary	This standard specifies the Milk thiamphenicol residue detection sample preparation, HPLC methods. This standard applies to milk thiamphenicol residues detected.

GB 29689-2013: Determination of Thiamphenicol residues in milk by High Performance Liquid Chromatographic method

---This is a DRAFT version for illustration, not a final translation. Full copy of true-PDF in English version (including equations, symbols, images, flow-chart, tables, and figures etc.) will be manually/carefully translated upon your order.
Determination of Thiamphenicol residues in milk by High Performance Liquid Chromatographic method National Standards of People's Republic of China National Food Safety Standard Determination of residual milk Thiamphenicol High performance liquid chromatography Published 2013-09-16 2014-01-01 implementation Ministry of Agriculture, People's Republic of China National Health and Family Planning Commission People's Republic of China released National Food Safety Standard Determination of residual milk Thiamphenicol High performance liquid chromatography

1 Scope

This standard specifies the sample preparation and high performance liquid chromatographic method Thiamphenicol milk residues detected. This standard applies to the detection of residual amount of milk thiamphenicol.

2 Normative references

The following documents for the application of this document is essential. For dated references, only applies to the version dated paper Pieces. For undated references, the latest edition (including any amendments) applies to this document. GB/T 6682 Water for analytical laboratory specifications and test methods Principle 3 The specimens remaining thiamphenicol, extracted with ethyl acetate, hexane degreasing, C18, purified by HPLC - UV determination, external standard The statutory amount.

4 Reagents and materials

The following reagents used, unless otherwise stated analytically pure, the water is in line with a water GB/T 6682 provisions. 4.1 thiamphenicol reference. content ≥99.0%. 4.2 ethyl acetate. 4.3 n-hexane. 4.4 Acetonitrile. chromatographically pure. 4.5 C18 solid phase extraction column.200mg/3mL, or equivalent person. 4.6 1mg/mL thiamphenicol standard stock solution. Weigh accurately thiamphenicol reference substance 10mg, in 10mL volumetric flask, dissolved in acetonitrile And dilute to volume, formulated at a concentration of standard stock solution 1mg/mL of. 2 ℃ ~ 8 ℃ storage period of one month. 4.7 10μg/mL thiamphenicol standard working solution. precise amount of 1mg/mL standard stock solution thiamphenicol 1.0mL, in an amount of 100mL Flask, diluted with mobile phase, formulated at a concentration of 10μg/thiamphenicol mL of standard working solution. Now with the current. 5. Apparatus 5.1 HPLC. with UV detector. 5.2 Analytical balance. a sense of volume 0.00001g. 5.3 Balance. a sense of the amount of 0.01g. 5.4 centrifuge. 5.5 SPE. 5.6 nitrogen blow instrument. 5.7 oscillating mixer. 5.8 heart-shaped bottle. 150mL. 5.9 rotary evaporator. 5.10 stopper plastic centrifuge tube. 50mL. 5.11 filter. 0.45μm. Preparation and Storage of sample 6 6.1 Preparation of the sample An appropriate amount of fresh or frozen or blank test milk and mix well. --- the test sample taken after homogenization, as the feed try. --- blank sample taken after homogenization, as a blank sample. --- blank sample taken after homogenization, adding a suitable concentration of the standard solution, is added as a blank sample. Save 6.2 sample Or less at -20 ℃. Determination Step 7 7.1 extract Sample Weigh 5g ± 0.05g, in 50mL centrifuge tubes, ethyl acetate was added 20mL, oscillation 10min, 4000r/min centrifugal 5min, the supernatant in the heart-shaped flask, the residue was added ethyl acetate 20mL, extraction was repeated once, the two extracts were combined, rotating at 45 ℃ Evaporated to near dryness, water was added to a 5mL flask heart-shaped, so that the ultrasonic 5min fully dissolved, transferred to 50mL centrifuge tubes. Add water to chicken 5mL After repeated dissolution vial heart go same centrifuge tube. 20mL hexane plus shaking 5min, 4000r/min centrifugal 2min, and the lower layer Liquid reserve. 7.2 Purification C18 column eluted sequentially with acetonitrile and water 5mL 5mL activation. Take stock solution through the column, controlling the flow rate 1mL/min, squeezed. 5mL acetonitrile , Collecting eluate, dry nitrogen at 45 ℃, the mobile phase the residue was dissolved 1.0mL, membrane filtration, for HPLC. 7.3 Preparation of standard curve The precise amount of 10μg/mL thiamphenicol appropriate amount of working standard solution, diluted with mobile phase to prepare a concentration of 20,50,125,250 and 500μg/L of the series of standard solution for HPLC assay. To measure peak area for the vertical axis, the standard solution concentration corresponding to a cross Axis, the standard curve. Seeking regression equation and correlation coefficient. 7.4 High Performance Liquid Chromatography 7.4.1 Chromatographic conditions 7.4.1.1 Column. C18 (250mm × 4.6mm, particle size 5μm), or equivalent person. 7.4.1.2 Mobile phase. aqueous acetonitrile (20 80, by volume), filtered through a filter membrane prior to use ultrasound. 7.4.1.3 flow rate. 1.0mL/min. 7.4.1.4 Detection wavelength. 225nm. 7.4.1.5 Column temperature. 30 ℃. 7.4.1.6 Injection volume. 20μL. 7.4.2 Assay Take a sample solution and standard solutions corresponding, for single or multi-point calibration, by external standard method, the peak area is calculated. Standard solution and sample solution Thiamphenicol fluid response of the instrument to be within the linear range of detection. In the chromatographic conditions, the standard solution and the blank test tissue was added FIG HPLC sample solution are given in Appendix A. 7.5 Blank test But without addition of the sample, the same steps employed in parallel operation.

8 and the result of the calculation expression

Residues thiamphenicol the specimens according to formula (1). X = c × V (1) Where. --- for X-try Residues feed thiamphenicol micrograms per kilogram (μg/kg); --- C concentration in the sample solution thiamphenicol, in units of nanograms per milliliter (ng/mL); V --- volume of the mobile phase the residue was dissolved in milliliters (mL); m --- try supply feed mass in grams (g). Note. The blank value should be subtracted from the results, expressed as the arithmetic mean of the measurement result measured parallel to three significant figures. 9 detection sensitivity, accuracy and precision 9.1 Sensitivity The detection limit of the method was 10μg/kg, the limit of quantitation of 10μg/kg. 9.2 Accuracy This method of adding 10μg/kg ~ 100μg/kg on recovery levels of 70% to 110%. 9.3 Precision The relative standard deviation of the method ≤15%, inter-assay relative standard deviation ≤15%.

Appendix A

Chromatogram Figure A.1 thiamphenicol chromatogram of the standard solution (50μg/L) Figure A.2 milk blank sample chromatogram Figure A.3 milk sample blank add thiamphenicol chromatogram (10μg/kg)

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