Some standard content:
IC557.040
National Standard of the People's Republic of China
GR/T 5009.28—2003
Replacement/5089
Determination of sacchar in sodium in foods
Delerminatior of sacchar in sodium in foods2003-08-11Promulgated
Ministry of Health of the People's Republic of China
Standardization Administration of the People's Republic of China
Implemented on 2004-01-01
GB/T5009.2B—2003
This standard replaces GB/T 9.281996°Determination of saccharin in foods 3. Compared with G/T 500S.2A—149, this standard has the following major modifications: The Chinese name of the standard is changed to "Determination of saccharin in food" in accordance with GB/T20014-2C01% standard Part 4: Chemical analysis methods. The original standard has been modified. This standard is proposed by the Ministry of Health of the People's Republic of China. The first method of this standard was drafted by the Food Safety Supervision and Inspection Institute of Dajin City, the Food Hygiene Supervision and Inspection Institute of Beining City, Wuhan City Health and Epidemic Prevention Station, Suzhou City Health and Epidemic Prevention Station, and Xichuan Province Health and Epidemic Prevention Station. The second method of this standard was drafted by the Health Bureau Food Safety Supervision and Inspection Institute. The third method of this standard was drafted by Shanghai Municipal Food Industry and Health Supervision and Inspection Institute, Taizhou City Health and Epidemic Prevention Station, Shanghai Nanshi District Health and Epidemic Prevention Station, Taiyuan City Health and Epidemic Prevention Station. This standard was first issued in 1565, revised in 1996, and revised for the second time. 224
1 Scope
Determination of saccharin sodium in food
This standard specifies the determination method of saccharin sodium in food. This standard is used to measure saccharin sodium in food. CB/T5009.28-2003
The detection limit of this method is 10% for high-efficiency liquid chromatography, 1% for sample volume, and 0.5g for detection volume: Method 1 High-performance liquid chromatography
2 Principle
Remove monohydrogen ether and ethanol from the sample, adjust the pH to near neutral, filter and conduct high-performance liquid chromatography, separate by reverse phase chromatography, and perform qualitative and quantitative analysis based on the peaks with retention time and area, 3 Reagents
3.1 Formaldehyde, filtered through a 0.5um filter.
3.2 Ammonia water and hydrogen water are mixed with an equal volume of water. 3.3 Acetic acid is dissolved (0.02 mul/1.), 1:4 k acetic acid is weighed, water is added to 1000 ml, and the solution is filtered through a 0.45 μm filter membrane. 3.4 Standard reserve solution of saccharin sodium, accurately take 0.551 g of sodium saccharin (CH3CONNaSO2I1,()) after drying at 120℃ for 14 hours: add water to 100 mL, the hot sodium content is 1.0 mg/mL, as the reserve concentration. 3.5 Standard reserve solution of saccharin sodium, take 10 ml of standard reserve solution of saccharin sodium, put it into a 100 ml volumetric flask, add water to the scale, filter through a u.45 nm filter membrane, set the filter band to be equivalent to 10 mg of saccharin 4.
High-performance liquid chromatography chromatograph, external detector. 5 Analysis steps
5.1 Sample treatment
5.1.1 Turn off the water; weigh 5.g~10.0 G g. and put it into a small cup, stir it at a low temperature to remove carbon dioxide, adjust the pH to about 7 with hydrogenated water (111), add water to make up to an appropriate volume, filter through a 0.4m sensitive membrane. 5.1.2 Fruit juice, weigh 5.0g~13g, adjust the pH to about 10 with hydrogenated water (1+1), add water to make up to an appropriate volume, precipitate, reduce the above and filter through a 0.45m sensitive membrane.
5.1.3 Prepare two kinds: Take 13.03 area - heat the water in a small calciner to remove the 7.0 sequence, adjust the pH with hydrogenated water 111? . Add water to make up to 20mL and filter through 0.45m filter membrane.
5.2 HPLC parameters
5.2.1: YWc C13 2, mm×210 mm: 0 um stainless steel. 5.2.2 Mobile rod alcohol acetic acid (0.02mol/L) 51SE). 5.2.3 Drop connection: 1rcl./m:n
5.2.4 Detection requirements: external detector, 23)mml wavelength.0).3A11FS. 5.3 Determination
Take the treated sensitive and standard samples and use a high performance commercial chromatograph (or equivalent) for separation. Use its standard drop of 225%
GB/T5009.28-2003
for qualitative analysis. Use its selected area to calculate the content of the measured substance in the sample: 5.4 Calculation of results
Calculate the potential content in the test sample according to the value. X
Where:
-saccharin content in the sample, in grams per kilogram [u)! The mass of the sample volume should be in grams (:
) The unit is m.
The total amount of the sample dilution is measured in milliliters (ml.) The result is calculated in grams (g).
5.5 The density
obtained under the condition of electrophilicity shall not exceed the absolute compensation of the arithmetic mean of 1 heart activity. 5.6 In addition, the high-efficiency phase separation device of 5.2 can be used to simultaneously measure the case of acid. Case acid and sodium azomethine, and its high-performance chromatography See,. Center
Mountain Road
Sugar Test
Figure 1 Commercial Color Range
Method 2 Thin Layer Chromatography
6 Principle
Under acid production conditions, sodium acetaldehyde is used to extract the essence in food, and the thin layer is used to extract the color. The color is then removed with a standard catalytic agent. For vacuum and semi-definite determination
7 Reagents
7.1 Ethyl acetate: no rapid oxidation.
7.2 Hydrothiol
7.3 Anhydrous alcohol: ethanol (55):
7. 4 Polyamide particles: n day
7.5 Hydrochloric acid (1-=): Take _00 m salt film, water to 260 ral..926
7.6 The developing agent is as follows;
7.6.1 Butanol nitrogen water + anhydrous ethanol (7+1+2). 7.6.2 Isoalcohol-ammonia water + ethanol water (7+1-2). GB/T 5009.28—2003
7.7 Color developer, read P with purple solution (0.4R/1.) Take 0.04R to replace neutral acid violet, dissolve with alcohol (50%, hydrogen peroxide fast solution (4g/L) 1.1mL to make pH 8. Make up to 1UmL: 7.8 Copper plaquenol solution (1002/[.]): Weigh 10× copper sulfate (CuS.-iH,0) and dissolve with water and dilute to 10nm: 1. 9 Chlorinated buffer (20g/L)
7.10 Saccharin sodium solution , accurately take (10X5g) sodium saccharin which has been dried for 4 hours, add ethanol to dissolve, put it into a 10ml volumetric flask, add ethanol (3.11ml to dilute to the mark, this solution is equivalent to 1mg sodium saccharin (CHCONVSO·2HO) per ml 8.1 Confirm paper: animal and post-transparent paper or non-concave paper 8.2 Please spray and other instruments.
8.3 Micro-injection syringe.
8.4 UV lamp: 253.7
8.5 Thin layer electrode r10 cm×20 cm or 20cm×20cm8.6 Development tank.
9 Analysis steps
9.1 Sample extraction
91.1 Beverage, ice, soda, take 11, =1. Homogenized sample (if the sample contains carbon dioxide, remove it first by adding viscous solution, if the sample contains argon, remove the oxidizing agent by heating in a boiling water bath) and put it in a 100-minute funnel, 2M acetic acid (111). Use 3C.20.25mL7.5% charcoal to extract once, combine 7.5% charcoal, wash once with 5mL of acidified water, and discard the water layer. After the acetaldehyde layer is dehydrated by anhydrous sodium sulfate, ethyl acetate is separated, and 2.0 μl of acetaldehyde is added to dissolve the residue, and the residue is stored in a sealed container. 1.2 Oil, fruit juice, etc.: weigh 20,000 ml or absorb 10 ml of the sample, put it in a 10 (L container, add water to about 63 mL, 20 ml of sulfuric acid concentrate (10UK/L), mix, and then add 1.1 ml of oxidizing agent (10 g/L), add water to the scale, mix, and stir. Stomach: 3min, pass 30ml. secretion when the 1250ml funnel, the following according to 9.1.1 self-salt <1F1) root method to break
9.1.3 solid fruit juice powder, etc.: weigh 20,000 ground uniform sample. Place in a ml. volumetric flask. Add 1ml. water, heat to dissolve, let cool, and follow 9.1.2 * add 20ml. grinding acid to make a concentrated solution (10℃g:1.) --\ and operate. 9.1.4 cakes, biscuits, etc. egg mouth.For starchy foods, place 25.0 g of the sample in a glassware with a suitable size, add 5% concentrated ortho-hydroxide solution (U.8 g/L). Tie the glassware tightly and place in a beaker with 20°C dialysis temperature (0.8 °C), add blood on the surface, dialyze overnight, collect 25 mL of the dialysis solution (equivalent to 12.5 g of the test solution), add about 0.4 mL of (1+) to neutralize, add 20 mL of copper phosphate solution (10 ug/L), add 4.4 mL of oxidizing solution (40/[.), mix, let stand for 30 minutes, and take 120 mL (equivalent to 10 g of the sample 1.) and place in a 2L separatory funnel. The following is obtained: 9.1.1 * 2 mL of hydrochloric acid (1-1)...\ 9.2 Preparation of thin layer plate
1. Amine powder, 0.4g powder, 7.0mL water, grind for 3min-5min, immediately apply to 3.25mm~3mr thick 10cm×23cm full layer, dry at room temperature, and store in a desiccator.
9.3 Spotting
2m below the full plate, use a syringe to spot 10 and 20L of sample solution at two points: 3.3, 5.0, ?.0, 1u.UrL sodium standard solution, the interval between the points is 1.5cm2
GB/T 50D9.282003
9.4 Development and Screening
Put the thin layer plate with the sample into a developing medium containing the developing agent (7.6.1 to 7.6.2). The developing agent should be about 0.5 cm thick and should have reached saturation in advance. Expand to 10 μm, take a small thin layer plate, spray it with the color developer, and the sample point should be yellow. Perform qualitative analysis based on the ratio of the sample point to the standard point, and perform semi-fixed determination based on the depth of the spot color. 9.5 Calculation
Calculate the weight of saccharin in the sample according to formula (2): X
Where:
×1000
The total amount of saccharin in the sample, in grams per dry gram or point per liter (g/kg or g/E), X
—the mass of saccharin in the sample for determination, in grams per dry gram or point per liter (g/kg or g/E), g>m
The amount or volume of the sample, in grams or milliliters (or L); the volume of the sample extract residue added, in milliliters (11.) V: the volume obtained at one point, in milliliters (mL). The third method is the determination method of sodium selective electrode
10 Principle
The sugar selective electrode is an electrode with FVC film made by quaternary plating as the sensing membrane. It is used in conjunction with the mercury electrode and the heptamercury electrode as the cathode to determine the content of saccharin in food. When the measuring temperature is low, the total ion intensity is consistent with the critical potential of the solvent, the measured potential follows the Nernst equation, and the potential difference changes with the activity (or concentration) of saccharin ions in the solvent. The content of saccharin sodium in the measured solvent is within the range of 0.02mE/ml.~)mg/mL. The electrode value is directly related to the negative logarithm of the concentration of the acetaldehyde:
11 Reagents
11.1 Acetaldehyde: saturate with hydrochloric acid (6 mpl/L) before use. 11.2 Anhydrous sodium hygrophosphate
11.3 Hydrochloric acid (6 TuV/.) Take 1 mm salt and dilute to 200 mL with water to make the aldehyde before use. 11.4 Sodium hydroxide solution (3.06 mol/L) is oxidized by 2.4R and the sample is diluted to 10001 ml. 11.5 System sensitivity reduction (1U3/L>): weigh 10% sulfur (CuS0,ITO) and add it to 190 mL water. 11.6 Caustic soda (15 g/1).
11.7 Hydrogenated paint solution (0.52 mol/1.) Dilute 1-.4. 11.B Sodium dihydrogen phosphate (NuH2PO4·2H2O>1mol/L) solution: Take 8g NaHPO2H2O, dissolve and transfer to 500ml empty volumetric flask, add water to dilute to scale, and adjust. 11.9 Disodium dihydrogen phosphate [e(NnHP-2H2O) 1mml/T solution, take 9.5g NaH1O·12HO in 250ml volumetric flask to dissolve, add water to dilute to scale, and adjust. 11.1C total high ion intensity dripping, %7. 1m sodium disodium phosphate solution (1mn1/1) 12.3 Disodium nitrate solution (1m/1) and mix to obtain.
11.11 Saccharin steel standard solution, accurately weigh 3.0351g, dry at 120℃ for 4b, transfer the saccharin solution to 1n3ml. Add olefin sieve, mix well and set aside. Each liter of this solution is equivalent to 1.r saccharin solution (1I, C(NNaS0-2HO) 12 Instruments
12.1 Acid meter or ion meter or other precision grade potentiometer, accurate to ±1mV228
12.2 Fast precision selective electrode.
GI/T 5009.28——2003
2.3217 type galvanostatic electrode: a double salt bridge galvanostatic electrode, the lower salt bridge contains 1% chloride solution (3um)/1.) 12.4 Agitator should be used,
12.5 Use effective glass paper for separation.
12. Semi-logarithmic paper.
3 Analysis stepsbzxz.net
13.1 Swab sample extraction
13.1.1 Liquid samples: concentrated wax fruit juice, juice, soda, soda, prepared slip, etc. 25.ml of sample (soda, sparkling wine, etc.) should be removed by carbon dioxide and then placed in a 250L bottle. Add 2L (6L/2) acid and add 5mL of saline solution once. Discard the water and transfer all the acetaldehyde to a 59mL container or bottle. Use a small amount of 7ml of freshly washed separating funnel to combine the original amount and use it to determine the acetaldehyde concentration. If necessary, add anhydrous sulfuric acid to remove the dehydrated food. 13.1.2 Foods containing high fat content: cakes and biscuits, vegetables, fried foods, etc. Weigh 23.W sample: place in a new bottle with paper towels, and adjust the efficiency of the bath ((2): 290 1. Put 1 mL of sodium hydroxide sieve (0.02 mL/L) in a beaker, add blood on the surface, dialyze 2111 mL, and occasionally stir the dialysate. Take 123 mL of dialysate, add about 0.4 mL of acid (0.5 mL/L) to make it uniform, add 20 mL of sulfuric acid to make it uniform, then add 14.1 mL of sodium hydroxide (40 g/L), and mix well. Let it stand for 30 minutes and filter. This 100mL filtrate is dried in a 250L full bucket, and the following or 13.1.1 "2mL with aldehyde (emal/I.) + \ liquid method detection. 13.1.3 dense hungry, weigh = 0.UV& accurate uniform sentence test sheet. Re-send to the city to increase the paper, add 5JmL sodium hydroxide drop liquid (J.ml/L): after mixing, the paper will be processed: sensitive person whole) certificate hydroxide dynamic wave (0.6mol/L> beaker, select analysis, sedimentation according to 3.1.3 frequency,
13.1.4 polished rice food: take 2 5. GCG is divided into small pieces of rice-like uniform sample, and the operation is carried out according to 1%. 1.2. 13.2 Determination 13.2.1 Preparation of standard solution: Accurately take 0, 0.5, 1.0.2.5, 5.0, 10.0mL sodium ions (corresponding to 0.0.5-0.0.2.5, 5.0, 10.0mg sodium ions), divide into 30mL volumetric bottles, add 5mL of total sodium ions to each, adjust the concentration, add water to the scale, and then: Put the sodium ions selection electrode and calomel electrode into the measuring instrument respectively. Connect the negative and positive ends of the electrode, insert the electrode into a beaker filled with water, adjust it to the operating state according to the user manual of the positioner, and add water to the starting position of the absorption under stirring (for example, the starting potential of a common electrode reaches 320mV). Use filter paper to collect the H electrode. Plot the above standard series of broken liquids one by one from low to high rates. Obtain the equilibrium potential (mV) during stirring.
On the semi-logarithmic oxygen, use density (mV>) as the frequency coordinate: the potential value (mV) is the coordinate. 13.2.2 Determination of the sample: Accurately draw 2mL 13.1 The soluble extract under item 13.1 needs to be placed in a 50mL filter cup and stirred until dry: add 5mL of total ionic strength adjustment solution by charcoal method, rotate carefully, shake and stir the cup to dissolve the residue, quantitatively transfer all the contents of the beaker into a 5L container, rinse the original beaker with a small amount of water several times, fill it to capacity, add water to the scale and mix evenly, determine its potential value (--mV) according to the law, and check the standard curve to obtain the number of grams of saccharin in the test solution. 13.3 Calculation of results: The content of saccharin is calculated by the formula <3, X=
A x1 co
Where:
X is the content of tantalum at the end of the sample, in grams per liter or grams per liter/liter or rt
GD/T5009.28—2003
4 is the amount of sodium extracted from the determination, in grams (mg) of acetyl alcohol extract.Unit: liter (L); V: the mass or volume of the sample after extraction with aldehydes, in units of mmol (m); m: the mass or volume of the sample, in units of grams (-nl). Result presentation: Same as 1, 1,
13.4 Interference
This method has no interference when the concentration of benzoic acid is between 200mg/kg and 100umg/kg: The concentration of sorbic acid is between 5)m/kg--0cmg: The content of saccharin sodium is within the range of 100g/kg~15mg/kg, and there is a positive error of about 3%~10%: Acid and hydroxyl radical have serious interference on the determination of this method. 2.u
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