Some standard content:
toS 62.040
National Standard of the People's Republic of China
GB/T 5009.169--2003
Determination of taurine in foods
Determination of taurine in foods2003-08-11Promulgated
Ministry of Health of the People's Republic of China
Standardization Administration of China
Implementation on 2004-01-01
GB/T5009.169—2003
This standard was proposed by the Ministry of Health of the People's Republic of China. The first drafting unit of this standard is: the Ministry of Health Food Hygiene and Epidemic Prevention Station; Hebei Provincial Epidemic Prevention Station, Shenzhen Baoan District Health and Epidemic Prevention Station. The first drafting unit of this standard is: the Ministry of Health Food Hygiene Inspection Institute; the supporting units are: Beijing Yiwu Small Epidemic Prevention Station, Handan Epidemic Prevention Station.
The second drafting unit of this standard is mainly composed of Zu Ying, Zhang Pingwei. GB/T 5009. 1692003
Taurine is a widely used taurine, with a chemical name of 2-aminoethyl ester. In 1993, my country approved the use of taurine in dairy products, infant food, beauty products and fortified beverages. This standard includes two methods: high-performance chromatography and thin-layer chromatography. 398
1 Scope
Determination of taurine in food
This standard specifies the determination method of taurine in food. This standard is applicable to the determination of taurine in beverages, infant food, cereal products and milk powder. GB/T5009.169—2003
The detection limit of the first method is 2U,0g: detection concentration 80ng/lg(L>, linear range: 0mR~C,Cim, the detection limit of the second method is 0.2m+ detection density 80m/k(L): The first method is high performance liquid chromatography
2 Principle
The sample is derivatized with a derivatizer after being absorbed, and the sample is separated by a C column and detected at its maximum yield at 330nm. The qualitative analysis is carried out according to the retention time and peak area. 3 Reagents
3.160x/1. Bacterial salicylic acid solution: Weigh 6,1 carbonyl salicylate, cation solution to 100mL. 3.2 phthalate [OP)
3.3 ethanol,
3.5 sodium hydroxide,
3.6 ethyl acetate,
3.7 sodium hydride.
3.B taurine: biochemical reagents,
3.9 0.4 mn/1. Turn over sodium borate solution, weigh 2.48g phthalic acid and 1.41g sodium hydroxide, dissolve in water to 100mL. 3.10 taurine, weigh 1.1 OPA and dissolve in 101nL methanol, add 0.1mT.Z alcohol, 0.1/L sodium borate solution to 100 3.11 Taurine standard solution: weigh 0.050 μg of taurine, dissolve it in water and add it to 50 mL of filter. Dilute the sample with water until it contains 1 μl of taurine. 3.12 Taurine standard solution: pipette 1 μl of taurine standard solution (3.11) into a 50 mL volumetric flask and add water to the volume to obtain 0.020 mg/mL taurine standard solution. 4.1 High-efficiency phase chromatography (with UV detector). 4.2 HPLC, 4.3 Wavelength cleaning device. 4.4 Volumetric flask: 5, 1 ml.,
4.5 micron port membrane filtration.
GE/5809.169—2003
5. Analysis steps
5.1 Sample treatment
5.1.1 Material: Accurately pipette 1, cm3 sample H water grid 101mL, wait for 5.1.2: Take 1.0g of pure. Make up to 25.0mL, pipette 3.0mL of the surface into 9 high centrifuges, add 3.0m60/. decomposing base water to correct acid. Centrifuge for 13min, pipette 2.0ml. Supernatant roll in 5mL discard plate bottle, add 1nol/t. sodium hydroxide to adjust the pH to neutral, and adjust to 5. (1 m1.. etc. 5.1.3 Preserved food: weigh 1.0# sample, add water to make up to 25.0ml, stir the sample thoroughly, let it stand for 5min, take 5.5ml of the above solution in a high temperature centrifuge, and adjust the following steps according to 1.2% water/30ml acid.. The operation is the same as that of milk products.
5.2.1 Derivatization, take 2mL of the above solution in a centrifuge, then accurately add 2.5% derivatization agent (3.10) and react for 2min. After 4min of milk filtration, perform PC analysis and determine the peak value. The sample standard should be injected at the same time and the injection should be kept consistent. The adjustment should be within 5min. The definite amount of taurine in the solution can be obtained from the standard curve. 5.2.21 FLC conditions
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Dynamic trace: Methyl alcohol + Art dark + system (t9+10+0) Wavelength: 330nm.1
Flow rate: m1.in
5.2.3 Standard curve: Absorb and mark respectively using 0, 0.5, 1.c, 2.5, 2.0..5tL. Then add water cart 2.5m1 below, 2.1\test accurately add 2.5mL of reagent, ...\start. Operation, then draw the graph, draw the standard curve or regression equation,
Color band medical observation Figure 1 quality,
Figure 1 6 Results Calculation formula: V, KI MU X: The content of foreign energy, in grams per dry gram / - - The selected liquid is calculated by risk, the unit is mg / mL) The total dilution volume, in milliliter a!: 00 V - the volume of the sample or the value of the total value of the difference in mass < Second method Thin chromatography method
The sample contains acid, after the exchange of the pseudo-pure, the full layer of chromatographic determination, quantitative. 8 Reagents
812% isocyanate: Accurately take 10.UmL of L saline, add 1 water to dilute to C; mL, 8.240/L sodium oxide filter, weigh 4 platinum hydride water curtain solution (12) m.8.32 stop: Analyze the purity.
3.4.1 Normal alcohol + ethyl acetate 5.22.2 + 0.8S.4.2 Normal alcohol + water + ethyl acetate <4 + 1 + 1) FB/35008.169-2003
8.% alkaline concentrated olefin exchanger: 717 barrier, use ethyl acetate to drain overnight, then rinse with water to remove color, strong irritant ethyl acetate cation exchange resin, 732 type, use single condensation to collect, then rinse with water until the water is colorless. 3u/] medium base fiber solution: weigh 0.3g whole methyl cellulose solution, add 11l of ethyl acetate, heat and dissolve, leave overnight, then reduce, take the filtrate for use.
8.8 Silica gel (2C month, for thin layer chromatography,
8.9 Color: weigh 0, 10g of the first: dissolve in 55m ethanol. 8.10 Nitrogen standard solution; take 0.0200g of the nitrophosphorus standard: dissolve in water and transfer into 1 30m volumetric flask, open and add water to the scale to release oxygen, and obtain 0.2mg/ml of nitroglycerin standard solution, 9 collection
9, 1 Analysis column: 1. 1 sm--30 em-
3.2 Heat I.
9.3 #5~20l
Excitation sampler: 10 pL.
9. 5. Lipid preparation
9,6. Water bath.bzxZ.net
9,7. Glass fog.
9.8. Hair dryer.
10.1. Preparation of ion exchange resin
Resistant to ion exchange: The most water-removed carbon anion is 717, and the 1.5cm~10cm layer must be exchanged. Do not mix with the gas. First, 5m. water (neutral effluent) is washed with water. 10.g: the effluent is neutral and the sodium hydroxide is passed through the layer. The sodium hydroxide is injected into the layer to be used as a standby.
10.2. Test treatment
10 1. The sample was evenly separated from the exchange resin by air flow, and the flow rate was adjusted to 30/m3. When the liquid level dropped to 431°C at the resin end, 25 mL of water was added for elution, and then 25 mL of 2% tantalum was added. The above two elutions were removed, and finally 2.2% tantalum was used for deionization. The sample was eluted with a rotary evaporator (temperature 59°C). 1.5 mL of water was added accurately to decompose the residue and transfer it to a test tube. This solution was prepared for thin separation. 10.2.2 Cereals: Take 2.0 were collected and extracted twice with 25, 10, and 7 ml of water respectively. Each extraction was allowed to stand for 15 min. The supernatant was transferred to the binary column with a pipette and the flow rate was adjusted to 50% disinfectant/mi. The column was rinsed with 5 ml of water. All the working liquid and water were exchanged. Note: The following exchange was carried out according to the "3% disinfectant/mi" in 1.2.1. Finally, add 1 ml of water accurately. 10.2.3 For powder, take 2.0g of uniform sample in a beaker, add 25ml of water, heat for 2min (do not let it boil), cool it down and transfer it to a vacuum hood, wash the beaker with 10ml of water, transfer the washing liquid to the vacuum hood, adjust the flow rate to 1/min, wait for the flow to reach the end, add 50tnL of water, press the selected temperature, and transfer it to a vacuum hood. Cation exchange. The following operations are in accordance with the "test flow rate 30ml/mi..." in 0.2.1. Therefore, accurately add 2.0mL of water barrier solution to make, filter the layer and perform the analysis. The used document is as follows: 10.3 Specifications 10.3.1 Preparation of thin layer plate Weigh 1: E gel, add 47ml. R/T. methyl cellulose group solvent (if too much, add appropriate amount), grind into 0.% m 10.3.2. Sample spotting
Place 3 points under the full layer plate, use a syringe to spot 2L of sample rate: at the same time spot 1.U.2.C, 3.0mL of iodic acid standard solution at one point, the distance between each point is 1m
10.3.3 Development and color development
Place the spotted layer plate in a sensitive environment and develop it to 3 points, 1,2, 2, 1, ... 11 Calculation of results
Calculate according to formula (2):
Wherein:
A×V×1000%100G
Y: Xm1000×1000
Amount of taurine, unit is per gram (kg): A
Amount of taurine in the sample, unit is microgram (g): Volume of water after evaporation in water bath, unit is ml (ml-sample volume, unit is microliter) 5
—Amount of sample, total unit is gram (ml) 1402
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