Some standard content:
1Ls67.040
National Standard of the People's Republic of China
GB/T5009.82—2003
B/1258—
Determination of vitamin A and vitamin E in foodsDetermination of residual vitamin E in foodsPromulgated on August 11, 2003
Ministry of Health of the People's Republic of China
Administrative Committee of Standardization of the People's Republic of China
Implementation on January 1, 2004
GR/T 5009.82—2003
This standard shall apply to the determination of vitamin A in infant formula—High-purity liquid strip method (CA), 1994 edition.
The first method in the standard is the colorimetric method for the determination of vitamin A in foods (1994 edition) of AOAC.71.2.
This standard is consistent with AOAC, S2.C6 and AOAC.574.2IV. The degree of consistency between this standard and AOAC is non-equivalent. This standard replaces 1/13F8-10 Method 3 for the determination of vitamin A and vitamin E in foods. Compared with GB/12388-990, the main adjustments of this standard are as follows: The Chinese name of the standard is revised. The Chinese name of the standard is revised. The structure of the original standard is revised. The standard is revised. GB/T20001.4-3001 Standard Compilation Rules Part 4: Chemical Analysis Methods. The standard is proposed and merged by the Ministry of Health of the People's Republic of China. Technical originator: Nutrition and Quality Research Institute of Chinese Academy of Defense Technology. The main originators of this standard are Shanghai Industry, Li Pin, and Tu Guodong. The standard is in! This standard is a small method for the determination of vitamin A and vitamin E in food. This standard is applicable to the determination of vitamin A and vitamin E in food. Standard limit: V: 0.8 g-E; -E 35.&206. Method 1: High performance liquid chromatography 2: Principle GR/T 5009.82—2003 3: After the final electrochemical extraction of the vitamin A and vitamin E in the sample, separate them into an organic solvent: High performance liquid chromatography, separate the vitamin A and vitamin E, and perform UV detection. Quantitative determination is performed using the internal standard method. 3: 3.1: Ether: Does not contain peroxides. 3.1.1: Peroxide inspection method: Use 5mL of 10% ether to drip, shake for 1min, if there is peroxide, free reaction will be released, the water layer will be colored or add + drops, add starch to the solution, the water layer will be colored. The acetaldehyde needs to be treated before use. 3.1.2 Method for removing peroxides: When re-distilling acetaldehyde, put pure 10% of the initial liquid in the bottle and discard 13% of the liquid:
3.2 Anhydrous acetaldehyde; it must not contain aldehydes. 3.2.1 Method, take 3mL of oxygen-limited solution and dissolve it in a test tube, add a little acetaldehyde or acetaldehyde, and evenly add oxygen to the solution. If there is a silver reaction after heating or cooling, it means that there is aldehyde in the acetaldehyde. 3.2.2 Decomposition method, take 2 nitric acid and silver and float them in a small amount of water. Add 4% sodium hydroxide to warm ethanol. Put the two old and old in ethanol, replace them, and place them in a dark place for a long time to stimulate the reaction (to promote the reaction), filter them, and store them in a hot storage bottle. Remove the 5mL of the distilled liquid, add appropriate amount of silver iodate, 3.3 anhydrous sodium sulfate:
3.4 ethanol, and use them after reheating.
3.5 Heavy water: add a little potassium hydroxide to the water, prepare before use: 3.6 Ascorbic acid solution (100g/L), prepare before use: 3.7 Potassium hydroxide solution: 1+T),
3.8 Nitrate solution (100g/).
3.9 Nitrate solution (50A/1)
3.10 Ammonia solution: add hydrogen water to nitric acid solution (3.5), until the generated precipitate is dissolved again, then add hydrogen solution 1.0 times full, if precipitation occurs, add water until it is 100% to the solution. 3.11 Vitamin A standard solution: safflower (6%) or ethyl acetate (pure water 0%>> after chemical treatment: use vitamin A standard product with dealdehyde ethanol, its degree depends on the concentration. About 1L is equivalent to 1 mol of retinol: Before use, use the spectrophotometer to calibrate its accuracy.
3.12 standard micro-biota: G-toxin purity S5%)-toxin (95%). Yogurt (95%). Deacetylation rate is 2.75%, 1 mol of hyaluronic acid (95%). Use UV spectrophotometer to calibrate the accurate concentration of these three vitamins and five melting points. 593
CH/T 5009.82—2003
3.13 Internal standard bag: weigh the purity of the product and prepare an internal standard with a ratio of 1. 14 εH_~14 test paper:
4 Fluoropolymer and equipment
4. Laboratory operation only.
4.2 High efficiency photometer with UV detector. 4.3 Heat radiator:
4.4 High speed centrifuge
4.4. 1 Small centrifuge tube; its plastic * 1. 5mL--3. 0 mL whole grid centrifuge (matching with high-speed centrifuge) 4.5 high-purity river gas,
4.6 city water bath
4.7 external spectrophotometer II.
5 analysis steps
5.1 sample treatment
5.1.1 chemistry: weigh 1~10) samples (containing vitamin A and vitamin F isomers for 4), ml, anhydrous ethanol, stir until the particles are uniform: m, 10 ascorbic acid, 2.0% phenyl ether, 10% acetic acid, 10% chlorinated benzene ... 5.1.3 Wash: Wash the acetaldehyde in the separatory funnel with about 5:1mL water, and press the PF test paper until the water layer does not decrease significantly (so wash lightly with water, the strength can be increased)
5.1.4 The concentrated acetaldehyde solution can be passed through a 25um sodium sulfate (about 5g) heat exchanger equipped with a 50ml flask. 0mmL ball evaporator, rinse with fermentation and anhydrous aldehyde for about 10 times, transfer the mixture to the evaporator, and then recover the 2 ethyl acetate in the bottle under pressure in a water bath. Remove the bottle and remove the mixture with a little air. Add 2 ml of ethanol immediately, mix thoroughly, and extract 51.5 ml of ethanol into a small plastic centrifuge tube (<4.4.1 ℃ ≤50001/min). 1 clear solution is then used for chromatographic analysis. If the content of vitamins in the sample is too low, it can be blown twice and then re-determined with a filter. 5.2 Preparation of standard curve 5.2. Standard concentration of vitamin A and vitamin E: Take 1 μl of the standard solution of each vitamin A and vitamin E and mix it in the following volume ratio. According to the formula, the absorbance of the vitamin at a given wavelength is deduced to be the absorbance coefficient of the vitamin. The determination conditions are shown in Table 1. Table 1
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Edodian formula (1)
But the standard curve
Comparative light absorption
Where:
The unit is
The average UV absorbance of the vitamin core drug is
A unit of the added value is micro; E
Specification 1% [Absorbance coefficient:
The model is diluted by times,
5.2.2 Preparation of standard curve
GB/T 5009.82—2003
This standard adopts the internal standard method. Mix a certain amount of vitamin A, tocopherol, tocopherol and internal standard liquid, and select whether to adjust the concentration so that the peak height of each injected substance is 70% of the full amount, which is the high concentration adjustment point, and the 1/2 is the low concentration point. The concentration of the internal standard benzodiazepine is not changed. The colorimetric analysis of the two concentrations is carried out. The color chart (Figure 1) is drawn. The ratio of the peak area of the vitamin A to the peak area of the internal standard is used as the effective standard. The vitamin A concentration is the standard. If there is a microprocessor device, it is determined by the one-point judgment method according to the instrument instructions. 4. 14
5.2 Yuan-D internal standard
S.AF cattle breeding
12.11α vitamin
Figure·vitamin A and vitamin F chromatogram
internal pile cannot separate HE and YF and change YK spines containing 3F samples.5.3 HPLC analysis
5.3.1 Chromatographic conditions (reference conditions)
5.3.1.1 Sample: ulrshere 10 m.4mm4, cm.5.3.1.2 New, ultrzsphere (>Ds4.6ux5.5.3.1.3 Mobile and: 1 ton of water 98-2, Mixing: Degas before use, 5.3.1.4 Ultraviolet detection wave K, 30am: or range 0.025.3.1.5 Sample: 2 1..
5.3..6t.tl. ml./mi..
5.4 After sample analysis
, take 20 ml of the modified vitamin solution and draw a color graph. After the color graph is changed, the remaining 5.4.! Empty: Use the cup to select the core to ensure the time of qualitative analysis 5.4.2 The root length of the sample is calculated to find out the ratio of the sample and the internal standard sample, and find out the value of the color standard online. Calculate the content of the vitamin
CE/T 5009.82—2003
5 The result is calculated in the formula,
The content of vitamins or, the unit is 100 grams nRo The unit is calculated by the standard curve and the unit is grams per rate (): the sample concentration is determined by volume, the unit is liter),
The sample is pressed on the plate, the unit is grams (g).
Let the calculation result be expressed to three significant figures
7 Precision
Repeatability The absolute difference between the two independent determinations shall not exceed 1% of the arithmetic mean. The second method is colorimetric method
8 Principle
Vitamin A interacts with a chemical matrix in chloroform to produce a blue substance which is proportional to the amount of vitamin A in the solution. The blue substance is not stable, but its absorbance at 62 nm can be measured within a certain period of time. 9 Reagents
Unless otherwise specified, only analytically pure reagents and water or appropriate pure reagents were used in the analysis. 9.1 Anhydrous sodium sulfate
9.2 Acetic anhydride
9.3 2-Hydroxyethyl alcohol
9.4 Anhydrous ethanol
3.5 Trichloromethane should not contain decomposition drugs: it will destroy the generated ions. 5.1 Hazardous changes: Trichloromethane is unstable and is easily affected by the air after being placed to generate hydrogen ions and phosgene: when testing, take a little water from the monochloromethane test tube and dissolve the hydrogen ions into the water layer. Add silver chromate solution and a white precipitate will be observed, indicating that there are decomposition products in the monochloromethane system.
9.5.2 Treatment method: The reducing agent should be free of decomposition products. If there are any, wash it several times with water, dehydrate it with anhydrous sodium chloride, and then evaporate it. 9.6 Dichloroethane solution (52R/1.>, prepare sodium chloride solution with chloroform, store in brown bottle (do not absorb decomposition).
9.7 Oxidizing agent (1-1)
98 Vitamin A or retinol acetate standard, prepare with 3.11. 5.2.1.9.9 Indicator (10R/T.). Prepare with 955% Z. 10. ||10. Experimental instruments.
10.2 Spectrophotometer:
10.3 System condensation tester,
11: The analytical step
Visor A is very easy to be destroyed by light, and the operation should be carried out under a microscope or with a glass collector. 11. Sample treatment
Depending on the test conditions, the bottom method or the grinding method can be used. 11.1.1 Chemical method
GD/T 5009.82—2003
Applicable to samples with different vitamin A content, reduce the time consumption of the whole test process, and easily cause vitamin A loss.
11.1.1.1 Chemical analysis: accurately weigh 0.5×10 samples or 1㎡ potassium hydroxide (111) and 20m-40m7 of saponification in a triangular flask, and saponify on a hot plate for 30min. 11.1.1.2 No matter how high the saponification rate is, separate the liquid from the mixture and wash it with 50m3 of water. The liquid can be filtered out and placed in a degreasing bucket. Wash twice with 5L of acetaldehyde, and then put it into a sliding bucket. According to the correct gas, set aside the layers, put the water layer into the first separatory funnel, and then rinse with ether twice. After washing, put the other end of the second separatory funnel, separate the layers quietly, add ether layer into the first separatory funnel, repeat until the water level is 5 times higher than that of the first separatory funnel.
11.1.1.3: Add 8L of water to the first separatory funnel, shake gently, clean the stomach, put away the water layer, add 15L-2℃.51el/hydroxylated butyl hydroxide to the separatory funnel, and after gently rinsing, remove the lower alkali solution to remove the acid-soluble soap. Continue to use 30mL of water each time. The main solution and the indicator are colorless (days) and set aside. m, be careful to remove the water,
11.1.1.4: put the aldehyde layer into a triangular bottle after anhydrous sodium sulfate, and then wash the material twice with about 25% acetaldehyde, and then put it into the bottle. The gastric water should flow up and absorb the acetaldehyde. When the remaining medicine in the bottle is 5.5, take it out and use vacuum to make the vitamin A content in the sample reach the appropriate range. 11.1.2 Grinding method www.bzxz.net
is suitable for the determination of the vitamin A content of samples greater than 5~10g, such as the analysis of the open. The steps are simple, the waste is simple, and the results are good.
11.1.2.1 Summary: The sample with a confirmed weight of 2R~55 is placed in anhydrous acid milling mill with a weight reduction of 5-5 times the sample weight, and the water in the sample is completely absorbed and homogenized. 11.1.2.2 Extraction: Carefully transfer all the sample to a sample bottle, add 50 mL to 109 mL acetaldehyde, and mix for 2 minutes to make the sample clear (11 to 2 minutes is required for large samples), or centrifuge to clear (because acetaldehyde is volatile, it should be operated in a cold water bath when the temperature is high, and the sample bottle containing acetaldehyde should also be immersed in cold water in advance) 11.1.2.3 Concentrate, take 2 mL to 5 mL of the acetaldehyde, put it into a colorimetric solution, and add 1 mL of chloroform to dissolve the residue immediately. 12. Preparation of standard lines Accurately take a certain amount of vitamin A standard Pour the solution into 45 volumetric bottles. Prepare the standard series with one drop of A, and then take the same amount of the standard series. Add 1 drop of B to each tube to make a standard colorimetric series: at n wavelength, adjust the absorbance to the horizontal point with one month, move the standard colorimetric series to the front of the light path in order, quickly add I sodium trioxide-trihydrogen alkane solution, determine the absorbance within 5 minutes, with the total absorbance as the ordinate and the vitamin content as the horizontal axis. Continue to prepare the standard curve,
12.2 Sample determination|| tt||Add 1 drop of 10m2 in one bottle and 1m3 in another bottle. Add 1m3 in the other bottles. Add 1m3 in the rest of the bottles. The sample liquid and 1m3 of acetic anhydride are the same as those in the standard line. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, international units are 1 unit per hundred) mg/100)
The content of vitamin A in the sample is checked from the standard curve in micrograms. The actual work is: 1. The test mass is in grams (g) in the first digit:
2. After adding clopine to the sieve, the volume is measured in a bottle, and the unit is (m/1n-1). The calculation result is guaranteed to be valid.
14 Precision
The difference between the results of two independent determinations obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.asa2. Calculate the ratio of the root length and internal standard sample, and find the value of the standard line. Calculate the content of CE/T 5009.82—2003. In the formula, the unit of vitamin C content is g/g: the sample concentration is expressed in liters, and the unit of vitamin C content is g/g: the sample concentration is expressed in liters, and the unit of vitamin C content is g/g.
Let the calculation result be expressed to three significant figures
7 Precision
Repeatability The absolute difference between the two independent determinations shall not exceed 1% of the arithmetic mean. The second method is colorimetric method
8 Principle
Vitamin A interacts with a chemical matrix in chloroform to produce a blue substance which is proportional to the amount of vitamin A in the solution. The blue substance is not stable, but its absorbance at 62 nm can be measured within a certain period of time. 9 Reagents
Unless otherwise specified, only analytically pure reagents and water or appropriate pure reagents were used in the analysis. 9.1 Anhydrous sodium sulfate
9.2 Acetic anhydride
9.3 2-Hydroxyethyl alcohol
9.4 Anhydrous ethanol
3.5 Trichloromethane should not contain decomposition drugs: it will destroy the generated ions. 5.1 Hazardous changes: Trichloromethane is unstable and is easily affected by the air after being placed to generate hydrogen ions and phosgene: when testing, take a little water from the monochloromethane test tube and dissolve the hydrogen ions into the water layer. Add silver chromate solution and a white precipitate will be observed, indicating that there are decomposition products in the monochloromethane system.
9.5.2 Treatment method: The reducing agent should be free of decomposition products. If there are any, wash it several times with water, dehydrate it with anhydrous sodium chloride, and then evaporate it. 9.6 Dichloroethane solution (52R/1.>, prepare sodium chloride solution with chloroform, store in brown bottle (do not absorb decomposition).
9.7 Oxidizing agent (1-1)
98 Vitamin A or retinol acetate standard, prepare with 3.11. 5.2.1.9.9 Indicator (10R/T.). Prepare with 955% Z. 10. ||10. Experimental instruments.
10.2 Spectrophotometer:
10.3 System condensation tester,
11: The analytical step
Visor A is very easy to be destroyed by light, and the operation should be carried out under a microscope or with a glass collector. 11. Sample treatment
Depending on the test conditions, the bottom method or the grinding method can be used. 11.1.1 Chemical method
GD/T 5009.82—2003
Applicable to samples with different vitamin A content, reduce the time consumption of the whole test process, and easily cause vitamin A loss.
11.1.1.1 Chemical analysis: accurately weigh 0.5×10 samples or 1㎡ potassium hydroxide (111) and 20m-40m7 of saponification in a triangular flask, and saponify on a hot plate for 30min. 11.1.1.2 No matter how high the saponification rate is, separate the liquid from the mixture and wash it with 50m3 of water. The liquid can be filtered out and placed in a degreasing bucket. Wash twice with 5L of acetaldehyde, and then put it into a sliding bucket. According to the correct gas, set aside the layers, put the water layer into the first separatory funnel, and then rinse with ether twice. After washing, put the other end of the second separatory funnel, separate the layers quietly, add ether layer into the first separatory funnel, repeat until the water level is 5 times higher than that of the first separatory funnel.
11.1.1.3: Add 8L of water to the first separatory funnel, shake gently, clean the stomach, put away the water layer, add 15L-2℃.51el/hydroxylated butyl hydroxide to the separatory funnel, and after gently rinsing, remove the lower alkali solution to remove the acid-soluble soap. Continue to use 30mL of water each time. The main solution and the indicator are colorless (days) and set aside. m, be careful to remove the water,
11.1.1.4: put the aldehyde layer into a triangular bottle after anhydrous sodium sulfate, and then wash the material twice with about 25% acetaldehyde, and then put it into the bottle. The gastric water should flow up and absorb the acetaldehyde. When the remaining medicine in the bottle is 5.5, take it out and use vacuum to make the vitamin A content in the sample reach the appropriate range. 11.1.2 Grinding method
is suitable for the determination of the vitamin A content of samples greater than 5~10g, such as the analysis of the open. The steps are simple, the waste is simple, and the results are good.
11.1.2.1 Summary: The sample with a confirmed weight of 2R~55 is placed in anhydrous acid milling mill with a weight reduction of 5-5 times the sample weight, and the water in the sample is completely absorbed and homogenized. 11.1.2.2 Extraction: Carefully transfer all the sample to a sample bottle, add 50 mL to 109 mL acetaldehyde, and mix for 2 minutes to make the sample clear (11 to 2 minutes is required for large samples), or centrifuge to clear (because acetaldehyde is volatile, it should be operated in a cold water bath when the temperature is high, and the sample bottle containing acetaldehyde should also be immersed in cold water in advance) 11.1.2.3 Concentrate, take 2 mL to 5 mL of the acetaldehyde, put it into a colorimetric solution, and add 1 mL of chloroform to dissolve the residue immediately. 12. Preparation of standard lines Accurately take a certain amount of vitamin A standard Pour the solution into 45 volumetric bottles. Prepare the standard series with one drop of A, and then take the same amount of the standard series. Add 1 drop of B to each tube to make a standard colorimetric series: at n wavelength, adjust the absorbance to the horizontal point with one month, move the standard colorimetric series to the front of the light path in order, quickly add I sodium trioxide-trihydrogen alkane solution, determine the absorbance within 5 minutes, with the total absorbance as the ordinate and the vitamin content as the horizontal axis. Continue to prepare the standard curve,
12.2 Sample determination|| tt||Add 1 drop of 10m2 in one bottle and 1m3 in another bottle. Add 1m3 in the other bottles. Add 1m3 in the rest of the bottles. The sample liquid and 1m3 of acetic anhydride are the same as those in the standard line. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, international units are 1 unit per hundred) mg/100)
The content of vitamin A in the sample is checked from the standard curve in micrograms. The actual work is: 1. The test mass is in grams (g) in the first digit:
2. After adding clopine to the sieve, the volume is measured in a bottle, and the unit is (m/1n-1). The calculation result is guaranteed to be valid.
14 Precision
The difference between the results of two independent determinations obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.asa2. Calculate the ratio of the root length and internal standard sample, and find the value of the standard line. Calculate the content of CE/T 5009.82—2003. In the formula, the unit of vitamin C content is g/g: the sample concentration is expressed in liters, and the unit of vitamin C content is g/g: the sample concentration is expressed in liters, and the unit of vitamin C content is g/g.
Let the calculation result be expressed to three significant figures
7 Precision
Repeatability The absolute difference between the two independent determinations shall not exceed 1% of the arithmetic mean. The second method is colorimetric method
8 Principle
Vitamin A interacts with a chemical matrix in chloroform to produce a blue substance which is proportional to the amount of vitamin A in the solution. The blue substance is not stable, but its absorbance at 62 nm can be measured within a certain period of time. 9 Reagents
Unless otherwise specified, only analytically pure reagents and water or appropriate pure reagents were used in the analysis. 9.1 Anhydrous sodium sulfate
9.2 Acetic anhydride
9.3 2-Hydroxyethyl alcohol
9.4 Anhydrous ethanol
3.5 Trichloromethane should not contain decomposition drugs: it will destroy the generated ions. 5.1 Hazardous changes: Trichloromethane is unstable and is easily affected by the air after being placed to generate hydrogen ions and phosgene: when testing, take a little water from the monochloromethane test tube and dissolve the hydrogen ions into the water layer. Add silver chromate solution and a white precipitate will be observed, indicating that there are decomposition products in the monochloromethane system.
9.5.2 Treatment method: The reducing agent should be free of decomposition products. If there are any, wash it several times with water, dehydrate it with anhydrous sodium chloride, and then evaporate it. 9.6 Dichloroethane solution (52R/1.>, prepare sodium chloride solution with chloroform, store in brown bottle (do not absorb decomposition).
9.7 Oxidizing agent (1-1)
98 Vitamin A or retinol acetate standard, prepare with 3.11. 5.2.1.9.9 Indicator (10R/T.). Prepare with 955% Z. 10. ||10. Experimental instruments.
10.2 Spectrophotometer:
10.3 System condensation tester,
11: The analytical step
Visor A is very easy to be destroyed by light, and the operation should be carried out under a microscope or with a glass collector. 11. Sample treatment
Depending on the test conditions, the bottom method or the grinding method can be used. 11.1.1 Chemical method
GD/T 5009.82—2003
Applicable to samples with different vitamin A content, reduce the time consumption of the whole test process, and easily cause vitamin A loss.
11.1.1.1 Chemical analysis: accurately weigh 0.5×10 samples or 1㎡ potassium hydroxide (111) and 20m-40m7 of saponification in a triangular flask, and saponify on a hot plate for 30min. 11.1.1.2 No matter how high the saponification rate is, separate the liquid from the mixture and wash it with 50m3 of water. The liquid can be filtered out and placed in a degreasing bucket. Wash twice with 5L of acetaldehyde, and then put it into a sliding bucket. According to the correct gas, set aside the layers, put the water layer into the first separatory funnel, and then rinse with ether twice. After washing, put the other end of the second separatory funnel, separate the layers quietly, add ether layer into the first separatory funnel, repeat until the water level is 5 times higher than that of the first separatory funnel.
11.1.1.3: Add 8L of water to the first separatory funnel, shake gently, clean the stomach, put away the water layer, add 15L-2℃.51el/hydroxylated butyl hydroxide to the separatory funnel, and after gently rinsing, remove the lower alkali solution to remove the acid-soluble soap. Continue to use 30mL of water each time. The main solution and the indicator are colorless (days) and set aside. m, be careful to remove the water,
11.1.1.4: put the aldehyde layer into a triangular bottle after anhydrous sodium sulfate, and then wash the material twice with about 25% acetaldehyde, and then put it into the bottle. The gastric water should flow up and absorb the acetaldehyde. When the remaining medicine in the bottle is 5.5, take it out and use vacuum to make the vitamin A content in the sample reach the appropriate range. 11.1.2 Grinding method
is suitable for the determination of the vitamin A content of samples greater than 5~10g, such as the analysis of the open. The steps are simple, the waste is simple, and the results are good.
11.1.2.1 Summary: The sample with a confirmed weight of 2R~55 is placed in anhydrous acid milling mill with a weight reduction of 5-5 times the sample weight, and the water in the sample is completely absorbed and homogenized. 11.1.2.2 Extraction: Carefully transfer all the sample to a sample bottle, add 50 mL to 109 mL acetaldehyde, and mix for 2 minutes to make the sample clear (11 to 2 minutes is required for large samples), or centrifuge to clear (because acetaldehyde is volatile, it should be operated in a cold water bath when the temperature is high, and the sample bottle containing acetaldehyde should also be immersed in cold water in advance) 11.1.2.3 Concentrate, take 2 mL to 5 mL of the acetaldehyde, put it into a colorimetric solution, and add 1 mL of chloroform to dissolve the residue immediately. 12. Preparation of standard lines Accurately take a certain amount of vitamin A standard Pour the solution into 45 volumetric bottles. Prepare the standard series with one drop of A, and then take the same amount of the standard series. Add 1 drop of B to each tube to make a standard colorimetric series: at n wavelength, adjust the absorbance to the horizontal point with one month, move the standard colorimetric series to the front of the light path in order, quickly add I sodium trioxide-trihydrogen alkane solution, determine the absorbance within 5 minutes, with the total absorbance as the ordinate and the vitamin content as the horizontal axis. Continue to prepare the standard curve,
12.2 Sample determination|| tt||Add 1 drop of 10m2 in one bottle and 1m3 in another bottle. Add 1m3 in the other bottles. Add 1m3 in the rest of the bottles. The sample liquid and 1m3 of acetic anhydride are the same as those in the standard line. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, international units are 1 unit per hundred) mg/100)
The content of vitamin A in the sample is checked from the standard curve in micrograms. The actual work is: 1. The test mass is in grams (g) in the first digit:
2. After adding clopine to the sieve, the volume is measured in a bottle, and the unit is (m/1n-1). The calculation result is guaranteed to be valid.
14 Precision
The difference between the results of two independent determinations obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.asa>, use chloroform to make sodium chloride solution, store in brown bottle (do not absorb the solution)
9.7 Oxidation solution (1-1)
98 Vitamin A or retinol acetate enzyme standard, 3.11. Co-selection method 5.2.1.9.9 Resistance indicator (10R/T.). Prepare with 955% Z. 10 Apparatus
10. Experimental routine Instruments. 10.2 Spectrophotometer: 10.3 System condensation tester. 11: Analytical steps: The sample is easily damaged by light, so the operation should be carried out under microscope or with a glass collector. 11. Sample treatment: Depending on the test conditions, the calorimeter method or grinding method can be used. 11.1.1 Chemical method: GD/T 5009.82—2003
Applicable to samples with different vitamin A content, reduce the time consumption of the whole test process, and easily cause vitamin A loss.
11.1.1.1 Chemical analysis: accurately weigh 0.5×10 samples or 1㎡ potassium hydroxide (111) and 20m-40m7 of saponification in a triangular flask, and saponify on a hot plate for 30min. 11.1.1.2 No matter how high the saponification rate is, separate the liquid from the mixture and wash it with 50m3 of water. The liquid can be filtered out and placed in a degreasing bucket. Wash twice with 5L of acetaldehyde, and then put it into a sliding bucket. According to the correct gas, set aside the layers, put the water layer into the first separatory funnel, and then rinse with ether twice. After washing, put the other end of the second separatory funnel, separate the layers quietly, add ether layer into the first separatory funnel, repeat until the water level is 5 times higher than that of the first separatory funnel.
11.1.1.3: Add 8L of water to the first separatory funnel, shake gently, clean the stomach, put away the water layer, add 15L-2℃.51el/hydroxylated butyl hydroxide to the separatory funnel, and after gently rinsing, remove the lower alkali solution to remove the acid-soluble soap. Continue to use 30mL of water each time. The main solution and the indicator are colorless (days) and set aside. m, be careful to remove the water,
11.1.1.4: put the aldehyde layer into a triangular bottle after anhydrous sodium sulfate, and then wash the material twice with about 25% acetaldehyde, and then put it into the bottle. The gastric water should flow up and absorb the acetaldehyde. When the remaining medicine in the bottle is 5.5, take it out and use vacuum to make the vitamin A content in the sample reach the appropriate range. 11.1.2 Grinding method
is suitable for the determination of the vitamin A content of samples greater than 5~10g, such as the analysis of the open. The steps are simple, the waste is simple, and the results are good.
11.1.2.1 Summary: The sample with a confirmed weight of 2R~55 is placed in anhydrous acid milling mill with a weight reduction of 5-5 times the sample weight, and the water in the sample is completely absorbed and homogenized. 11.1.2.2 Extraction: Carefully transfer all the sample to a sample bottle, add 50 mL to 109 mL acetaldehyde, and mix for 2 minutes to make the sample clear (11 to 2 minutes is required for large samples), or centrifuge to clear (because acetaldehyde is volatile, it should be operated in a cold water bath when the temperature is high, and the sample bottle containing acetaldehyde should also be immersed in cold water in advance) 11.1.2.3 Concentrate, take 2 mL to 5 mL of the acetaldehyde, put it into a colorimetric solution, and add 1 mL of chloroform to dissolve the residue immediately. 12. Preparation of standard lines Accurately take a certain amount of vitamin A standard Pour the solution into 45 volumetric bottles. Prepare the standard series with one drop of A, and then take the same amount of the standard series. Add 1 drop of B to each tube to make a standard colorimetric series: at n wavelength, adjust the absorbance to the horizontal point with one month, move the standard colorimetric series to the front of the light path in order, quickly add I sodium trioxide-trihydrogen alkane solution, determine the absorbance within 5 minutes, with the total absorbance as the ordinate and the vitamin content as the horizontal axis. Continue to prepare the standard curve,
12.2 Sample determination|| tt||Add 1 drop of 10m2 in one bottle and 1m3 in another bottle. Add 1m3 in the other bottles. Add 1m3 in the rest of the bottles. The sample liquid and 1m3 of acetic anhydride are the same as those in the standard line. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, international units are 1 unit per hundred) mg/100)
The content of vitamin A in the sample is checked from the standard curve in micrograms. The actual work is: 1. The test mass is in grams (g) in the first digit:
2. After adding clopine to the sieve, the volume is measured in a bottle, and the unit is (m/1n-1). The calculation result is guaranteed to be valid.
14 Precision
The difference between the results of two independent determinations obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.asa>, use chloroform to make sodium chloride solution, store in brown bottle (do not absorb the solution)
9.7 Oxidation solution (1-1)
98 Vitamin A or retinol acetate enzyme standard, 3.11. Co-selection method 5.2.1.9.9 Resistance indicator (10R/T.). Prepare with 955% Z. 10 Apparatus
10. Experimental routine Instruments. 10.2 Spectrophotometer: 10.3 System condensation tester. 11: Analytical steps: The sample is easily damaged by light, so the operation should be carried out under microscope or with a glass collector. 11. Sample treatment: Depending on the test conditions, the calorimeter method or grinding method can be used. 11.1.1 Chemical method: GD/T 5009.82—2003
Applicable to samples with different vitamin A content, reduce the time consumption of the whole test process, and easily cause vitamin A loss.
11.1.1.1 Chemical analysis: accurately weigh 0.5×10 samples or 1㎡ potassium hydroxide (111) and 20m-40m7 of saponification in a triangular flask, and saponify on a hot plate for 30min. 11.1.1.2 No matter how high the saponification rate is, separate the liquid from the mixture and wash it with 50m3 of water. The liquid can be filtered out and placed in a degreasing bucket. Wash twice with 5L of acetaldehyde, and then put it into a sliding bucket. According to the correct gas, set aside the layers, put the water layer into the first separatory funnel, and then rinse with ether twice. After washing, put the other end of the second separatory funnel, separate the layers quietly, add ether layer into the first separatory funnel, repeat until the water level is 5 times higher than that of the first separatory funnel.
11.1.1.3: Add 8L of water to the first separatory funnel, shake gently, clean the stomach, put away the water layer, add 15L-2℃.51el/hydroxylated butyl hydroxide to the separatory funnel, and after gently rinsing, remove the lower alkali solution to remove the acid-soluble soap. Continue to use 30mL of water each time. The main solution and the indicator are colorless (days) and set aside. m, be careful to remove the water,
11.1.1.4: put the aldehyde layer into a triangular bottle after anhydrous sodium sulfate, and then wash the material twice with about 25% acetaldehyde, and then put it into the bottle. The gastric water should flow up and absorb the acetaldehyde. When the remaining medicine in the bottle is 5.5, take it out and use vacuum to make the vitamin A content in the sample reach the appropriate range. 11.1.2 Grinding method
is suitable for the determination of the vitamin A content of samples greater than 5~10g, such as the analysis of the open. The steps are simple, the waste is simple, and the results are good.
11.1.2.1 Summary: The sample with a confirmed weight of 2R~55 is placed in anhydrous acid milling mill with a weight reduction of 5-5 times the sample weight, and the water in the sample is completely absorbed and homogenized. 11.1.2.2 Extraction: Carefully transfer all the sample to a sample bottle, add 50 mL to 109 mL acetaldehyde, and mix for 2 minutes to make the sample clear (11 to 2 minutes is required for large samples), or centrifuge to clear (because acetaldehyde is volatile, it should be operated in a cold water bath when the temperature is high, and the sample bottle containing acetaldehyde should also be immersed in cold water in advance) 11.1.2.3 Concentrate, take 2 mL to 5 mL of the acetaldehyde, put it into a colorimetric solution, and add 1 mL of chloroform to dissolve the residue immediately. 12. Preparation of standard lines Accurately take a certain amount of vitamin A standard Pour the solution into 45 volumetric bottles. Prepare the standard series with one drop of A, and then take the same amount of the standard series. Add 1 drop of B to each tube to make a standard colorimetric series: at n wavelength, adjust the absorbance to the horizontal point with one month, move the standard colorimetric series to the front of the light path in order, quickly add I sodium trioxide-trihydrogen alkane solution, determine the absorbance within 5 minutes, with the total absorbance as the ordinate and the vitamin content as the horizontal axis. Continue to prepare the standard curve,
12.2 Sample determination|| tt||Add 1 drop of 10m2 in one bottle and 1m3 in another bottle. Add 1m3 in the other bottles. Add 1m3 in the rest of the bottles. The sample liquid and 1m3 of acetic anhydride are the same as those in the standard line. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, international units are 1 unit per hundred) mg/100)
The content of vitamin A in the sample is checked from the standard curve in micrograms. The actual work is: 1. The test mass is in grams (g) in the first digit:
2. After adding clopine to the sieve, the volume is measured in a bottle, and the unit is (m/1n-1). The calculation result is guaranteed to be valid.
14 Precision
The difference between the results of two independent determinations obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.asaPreparation of standard colorimetric lines
Accurately take a certain amount of vitamin A and put it into 45 volumetric bottles. Prepare the standard series with one bottle of A, and then take the same amount of vitamin A. Add 1 drop of the standard series into each bottle of B to make a standard colorimetric series: at the wavelength of n, adjust the absorbance by 1 point, move the standard colorimetric series to the front of the light path in order, quickly add sodium trioxide-tris(III) solution, determine the absorbance at 5 °C, and use the total absorbance as the ordinate and the vitamin A content as the horizontal axis to prepare a standard curve.
12.2 Sample determination
Add 1 drop of 10 ml of vitamin A into one bottle and 1 ml of vitamin B into the other colorimetric bottle. Add 1 ml of the remaining colorimetric bottles respectively. The sample drop and 1 ml of acetic anhydride are the same as the standard. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, 1 international unit 1 total element! The unit is set grams per hundred (mg/1oo)
Check the content of vitamin A in the sample from the standard curve, the unit is microgram: the test mass, the first digit is grams (g):
After collecting, add clopine sieve to the volume of the bottle, the single digit is a pen (m2 1n-change each gram design.
Calculation results guarantee a single valid value.
14 Precision
The difference between two independent determination results obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.Preparation of standard colorimetric lines
Accurately take a certain amount of vitamin A and put it into 45 volumetric bottles. Prepare the standard series with one bottle of A, and then take the same amount of vitamin A. Add 1 drop of the standard series into each bottle of B to make a standard colorimetric series: at the wavelength of n, adjust the absorbance by 1 point, move the standard colorimetric series to the front of the light path in order, quickly add sodium trioxide-tris(III) solution, determine the absorbance at 5 °C, and use the total absorbance as the ordinate and the vitamin A content as the horizontal axis to prepare a standard curve.
12.2 Sample determination
Add 1 drop of 10 ml of vitamin A into one bottle and 1 ml of vitamin B into the other colorimetric bottle. Add 1 ml of the remaining colorimetric bottles respectively. The sample drop and 1 ml of acetic anhydride are the same as the standard. GB/T5009.82-2D33
13 Calculation of results
X=VTGC
Test result (such as international units, 1 international unit 1 total element! The unit is set grams per hundred (mg/1oo)
Check the content of vitamin A in the sample from the standard curve, the unit is microgram: the test mass, the first digit is grams (g):
After collecting, add clopine sieve to the volume of the bottle, the single digit is a pen (m2 1n-change each gram design.
Calculation results guarantee a single valid value.
14 Precision
The difference between two independent determination results obtained under the conditions of accuracy shall not exceed % of the arithmetic half value.
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