Testing of natural vitamin E in food additives

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A. 1 General regulations

Unless otherwise specified in this standard, the purity of the reagents used should be above analytical purity. The standard titration solution used, the standard solution for impurity determination, the formulation, and the product should be prepared in accordance with the provisions of GB/T601, GB/T602, and GB/T603. The experimental water should comply with the provisions of Grade III water in GB/T6682-2008. The solution used in the experiment refers to aqueous solution when no specific solvent is specified for preparation

A.2 Identification Test

A.2.1 Solubility

Insoluble in water, easily soluble in ethanol, and miscible in ether

A.2.2 Specific Rotation Test

A.2.2.1 Reagents and Materials

Trichloromethane: Chromatographic purity

A.2.2.2 Instruments and Equipment

Polarimeter

A.2.2.3 Analysis Steps

Weigh 5.0g of the sample, dissolve it in chloroform, and dilute it to 50mL. Place the sample solution in a polarimeter for determination, and follow the specific analysis steps specified in GB/T613.

. The measured specific rotation am (20 ℃, D) should be 0 (o) dm2 · kg-1 ± 0.05 (o) dm2 · kg-1

A.2.3 Nitric acid color test

A.2.3.1 Reagents and materials

A.2.3.1.1

; Anhydrous ethanol

A.2.3.1.2 Nitric acid

A.2.3.2 Analysis Steps

Weigh about 10mg of the sample, dissolve it in 10mL of anhydrous ethanol, add 2 mL of nitric acid, shake well, and heat in a 75 ℃ water bath for 15 minutes. The solution should appear bright red to orange

A.2.4 Maximum absorption wavelength test

A.2.4.1 Instruments and equipment

UV spectrophotometer

A.2.4.2 Analysis Steps

Weigh 0.01g of the sample, dissolve it in anhydrous ethanol and dilute to 200mL, shake well, and place it in a UV spectrophotometer for scanning. There should be a maximum absorption peak at 292nm ± 2nm.

A.3; Determination of vitamin E (dl-a tocopherol) content

A.3.1 Method summary

Gas chromatography is used to separate the sample from impurities under selected working conditions using a non polar quartz capillary column, and hydrogen flame ionization detector is used for detection. Internal standard method is used for quantification to calculate the content of vitamin E (dl-a tocopherol) in the sample

A.3.2 Reagents and Materials; N-hexane: chromatographically pure

A.3.2.2; Hexadecanol hexadecylate

A.3.2.3 Vitamin E (dl-a tocopherol) standard

A.3.2.4; Vitamin E (dl- α- Acetic acid tocopherol standard

A.3.3 Instruments and Equipment

Gas chromatograph, equipped with a hydrogen flame ionization detector.

A.3.4 Reference chromatographic conditions

A.3.4.1 Chromatographic column: column length of 30m, column inner diameter of 0.25mm, fixed phase of polydimethylsiloxane, and membrane thickness of 0.25um

A.3.4.2 Column temperature: 280 ℃

A.3.4.3 Detector: Flame ionization detector, temperature 290 ℃

A.3.4.4 Injection port: temperature 290 ℃; Split injection, with a split ratio of 1:30; Injection volume 1uL

A.3.4.5 Carrier gas: Nitrogen, flow rate 1mL/min

A.3.4.6 A.3.4.1~A.3.4.5 are recommended chromatographic columns and typical chromatographic operating conditions. Please refer to Appendix B for typical gas chromatography diagrams for dl-a tocopherol content determination. Other chromatographic columns and conditions that can achieve equivalent separation effects can be used

A.3.5 Analysis Steps

A.3.5.1 Preparation of Internal Standard Solution

Weigh an appropriate amount of hexadecyl acetate (internal standard substance), dissolve, dilute, and shake well with n-hexane, and use it as the internal standard solution.

. This internal standard solution contains 3mg of hexadecyl acetate per milliliter

A.3.5.2 Preparation of Standard Solution

Weigh approximately 30mg of vitamin E (dl-a tocopherol) standard to the nearest 0.0001g, place it in a 10mL brown volumetric flask, dissolve and dilute the internal standard solution to the mark, shake well, and use it as the standard solution

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