This standard specifies the determination method of carotene in food - high performance liquid chromatography and paper chromatography. This standard is applicable to the determination of carotene in food. The detection limit of this method: 5.0 mg/kg (L) for high performance liquid chromatography, linear range of 0 mg/L ~ 100 mg/L; 0.11 μg for paper chromatography, linear range of lng ~ 20ng. GB/T 5009.83-2003 Determination of carotene in food GB/T5009.83-2003 Standard download decompression password: www.bzxz.net

1C5 67.040
National Standard of the People's Republic of China
GB/T5009.83—2003
Generation H/T12389-.1S9
Determination of carotene in food
Delerminatinn nf carotcnc in foods2003-08-11 issued
Ministry of Health of the People's Republic of China
China National Standardization Administration
2004-01-01 implementation
This standard GB/1238913302 Standard for determination of hydroxyl radicals in foods: Compared with BT1238V-1990, this standard has modified the following Chinese names of the standard. The Chinese name of the standard has been changed to Standard for determination of hydroxyl radicals in foods: Add high performance liquid phase chromatography as a method; GB/15009.85-2003
According to GB/120001.4-2001 standard: Part 4 The chemical analysis method has been modified to the original standard: The production standard is issued by the Ministry of Health of the People's Republic of China and is the second standard. The first standard is drafted by the Ministry of Health and the third standard is exempted from the third standard. Beijing Municipal Health and Safety Station and the National Institute of Environmental Protection of the Chinese Academy of Medical Sciences are responsible for drafting. The first standard is drafted by Zuying Xiu, Li Anxue, Zhang Shanping, Sun Yang, and Xiaoyuan She. The second standard is maintained by the National Institute of Preventive Medicine of the Chinese Academy of Sciences. The second standard is drafted by Zhao Zhonglin, Xia Guangye, Gan, and Xia Rundong: The original standard was issued in 1910 and is the first revision. ul
CH/T 5009.83—2003
It is an important nutrient for human body. It is the body of vitamin A and is an important component of food. GH is equivalent to 10 vitamins. The country has approved 3 of them to be added to oil and chemical foods, and it is allowed to be added to 14 kinds of foods such as human beverages, butter, ice cream, etc. In 1993, it was approved as a nutritional enhancer for children's food. In 2009, the National Food and Drug Administration (GB/T 12009-1999) formulated the method for determining the nutritional value of carotene in food. This method proposes a rapid and effective method for determining the nutritional value of carotene in food by color analysis, which is the first method. The method for determining the nutritional value of carotene in food (GB/T 12009-1999) is the first method. Scope
Determination of carotene in food
This standard specifies the determination of carotene in food by high performance liquid chromatography and chromatographic method. This standard is applicable to the determination of carotene in food. B/T5009.83—2003
The detection limit of this method is: HPLC step.0/g>, the sensitivity range is m/~/, the chromatographic method is .11, and the linear range is 120ng
The first method is high performance liquid chromatography
2 Principle
2 Carotene in the sample is extracted with sodium hydroxide (8C-2J), spheroidized with zirconium trioxide, and then HPLC method is used for qualitative analysis by retention time and quantitative analysis by peak height or area
3. 1. Aldehyde boiling range ~ic.
3.2 Methanol: color potential pure
3.3 Propanol.
3.4 ​​Alkane.
3.5 Tetrahydrofuran,
3.6 Methane.
3.7 Ethylene, chromatographic line.Www.bzxZ.net
3.8 Trichloroethane, 1) 2 days, 140 ℃ 22, take out and put into the concave beam to dry. 3.9 Containing alkane: take out and dry 1n1, dissolve with isooctane and dilute to 251nL, take out and add 3.10-Carotene, add one less 1-carotene in the beaker, dissolve in the beaker, then wash with petroleum solvent several times, transfer to the maximum volume, fixed rate, fill this with /m, store in 8 mouths. 3.11 The following standard is missing: accurately weigh 3-12.5 ml of alkane in a cup, first, add a small amount of alkane, then extract with oil and add the whole cup. The amount of alkane is not transferred to the 3-10ml cup, and the concentration is 250/mL, which can be stored for 13 days. Stable within two months: according to the concentration limit of the 3-10ml standard, the mobile phase is 1g/mL. 3.12 The following standard is effective: respectively absorb the standard (5, 1, 0.2.C.3.3.4.0.5.0) in a 10ml bottle, add the mobile phase to the scale, and then get the standard 5, 10, 20, 40g/mL of alkane.
3.139-The following standard is effective: you take!, Place the mixture in a 10-well bottle, fill it with 10 ml of isooctane melt containing oxygen, and irradiate 5 ml of isooctane at a distance of 3 cm from the light source. Then remove the solvent in a vacuum. Decompose the isooctane with a small amount of oxygen, and use a constant volume solution to a concentration of 5/13, and store it. 4 Apparatus
4.1 High performance liquid chromatography,
4.2 Centrifuge.
4.3 Rotary evaporator,
GB/I5009.83-200 3
5 Analysis steps
5.1 Extraction
5.1.1 Type of food: 10.Cg total sample + 25mL cut off from a small tube with a stopper or a commercially available small tube (at least more sample can be obtained) with a cotton ball or ether (8V+2C>) mixed with a full-area cotton ball, and then absorbed in a colorless bag and retained in a special evaporator until the liquid is colorless. The extract is evaporated to dryness in a rotary evaporator (water temperature 30℃) 5.1.2 Liquid food: 10.C mL test Y at 25)T. Add 20ml of acetone (80-2U) dissolved in oil. Extract, then statically separate the layers, divide the lower layer into 4 parts by gradual concentration, until the extract is colorless, combine the extracts, evaporate on a rotary evaporator until the water solubility is 40.5g/cm2, add sodium methacrylate (8C+20), take out, and extract the upper layer with a single white color, and obtain the safety data. Evaporate on a rotary evaporator at a full recovery rate:. 5.2 Shear the chemical
,, 1.5.1.2,, 1.3 in the manner of old liquid, use a small amount of sodium sulfate to dissolve the whole, sign and perform folding: the alumina layer is 1.5cm (inner diameter) and 1cm (outer diameter), first use eluent (5 + 95>) to wash the alumina, then add the falling liquid into the competitive group, inject the elution liquid at a constant volume of 1 minute, the flow rate is 2 drops/rim, collect the eluate on a 3.45mm pore filter membrane, and use PLC analysis. 5.3 Determination 5.3.1 HPLC column: Spheriso column: 4.Smr.×15Jnur.r mobile phase, methanol-methanol (S). Mobile phase: 1.7f./i-1m. 5.3.2 Detailed determination: Absorb 20L of the chemical solution in 5.2 according to the standard and obtain the maximum content of the three elements from the standard curve. 5.3.3 Standard curve: Take 29L of the standard solution respectively and carry out HPLC analysis. Use the area below to mark the abundance. Calculate the result (see Figure 6).
VX×*00n×1cC8*1 00
Test-measurement unit: grams per liter) or /
A certain amount of wear, single piece is open (L)
\——The concentration of 8 elements in the sample (obtained on the mark), the unit is microliters per liter (tons/1.) Amount of sample, single test is grams (actual liter) m section nesting South two effective digits.
7 Degree of sensitivity
In the case of repeated testing, the absolute difference of the results obtained in the second independent test cannot exceed the arithmetic mean of 1,ut
8 Original leech
The second method is paper screen analysis method
GB/T 5009.83—2003
After the sample is chemically treated, use a slurry to extract the chlorine in the food and other substances. Use petroleum aldehyde as the developing agent to carry out the paper layer. Cut off the polarity of the sample and move the fastest. Cut off the required zone from the negative electrode. Elute it in 50m9 reagent
9.1 Petroleum aldehyde (boiling point ~60%). 9.2 Hydroxide solution (1+1, 50% potassium hydroxide solution. Water. 9.3 Anhydrous ethyl acetate. It must not contain any other substances. 9.3.1 Method,
9.3.1.1 Silver hydrogenation: Add concentrated water to 10% nitric acid solution, fully emulsify and precipitate, add a few drops of 2.5mu/mol sodium hydroxide. If a precipitate is formed, add concentrated water to reduce it. 9.5.T.2 Silver can react, add 2 root chlorine solution, add a few ethanol, add at least 2.5mol/l of molten metal and heat mechanically. If there is no aldehyde in the ethanol, it means there is no precipitation. If there is no aldehyde in the ethanol, it means there is no precipitation. 9.3.2 The aldehyde removal method is achieved. Take 2x the correct root and add 4 sodium hydroxide with a small amount of water. Pour the two into 7 wells and place them in the stomach for two days (from time to time to promote the reaction). Filter and steam in a can. Discard the left and right sides. When the ethanol contains more aldehydes, the amount of aldehyde can be appropriately increased.
9.4 Anhydrous aldehyde.
9.5 β Preparation of standard solution
9.5.1 Prepare the standard solution accurately and weigh 55.0mg. Prepare the standard solution with 1m.m. Trichloroethane method and accurately measure the total concentration of about 500/ml. The calibration concentration is as follows: Take the standard solution (10). Process with 3.00T of ethane. Measure the absorbance value, the original colorimetric cup is 1 cm, and the blank is n-hexane. The average value is determined by the formula (gate):
Wu Zhong:
X, the standard liquid concentration is gram per liter (I) A
Love light value
The wavelength of the incident light in the F mountain is 4503m, the original colorimetric cup is 1cm, and the solvent range is! m/. The absorption coefficient is 0.263. The conversion coefficient is taken from the determination process. 9.5.2 The following procedures are used: The standard solution for special calibration is accurately diluted 10 times with the test solution, and the increase is equal to 5 stations of light water and stored in ice!
Note: In some cases, the standard cannot be completely dissolved in the organic solvent. If necessary, the standard should be electrolyzed first, then extracted with organic solvent, and then concentrated to neutral with distilled water. Then calibrate. Because the standard is very easy to decompose, the standard should be calibrated before each use. When measuring the sample, the standard should be used synchronously. 15 Instruments
10.1 Experimental equipment.
iB/1 5009.83—2303
10.2 Diffraction analysis
10.3 Spectrophotometer:
10.4 Rotary igniter with 150L speed bottle. 10.5 Water absorption error:
10.6 Adjust the sample,
10.? Sample tip or injection point.
10.2 Sample pretreatment
11. 1. Saponification
Take an appropriate amount of sample, mix it with the original sample: the amount of carotene content is about 1~25 mL, the amount of food oil and high fat sample should not exceed 13%, add 1 mL of dealdehyde alcohol into a standard bottle, then cover with 111mL of potassium oxide, heat in a concave flow for 30 minutes, and then cool it rapidly with water. After electrochemical extraction, the sample is extracted with aldehyde until the amount of aldehyde is reduced to E. The amount of aldehyde used for each extraction is 15 mL-25 mL. Washing
Put the treated sample into the water, add the extracted solution through a small filter with anhydrous sodium sulfate, filter it into a spherical ball, inject the color layer into the centrifuge and cool it several times. 1.1.3 Reduction and volume adjustment
Put the extract in the upper or lower flask on an evaporator with reduced pressure and set the temperature to 0.5 °C. Replace the evaporator with nitrogen for 1 hour, collect the spherical newspaper, blow nitrogen for 10 minutes, and immediately add 2.5% ether to make it stable. 1.2 Paper chromatography
11.2.1 Stop at 4 °C. Place the bottom of the 9m:3m filter paper on the bottom.
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