Product name: Vitamin E Assay Kit

Catalog number: E-BC-K033

Detection method: Colorimetric method 

Detection instrument: Spectrophotometry (Visible range)

Valid period: 3 months 

Experimental instrument

Glass tube, Micropipettor, Vortex mixer, Centrifuge, Spectrophotometer (533 nm wavelength)

Application

This kit can be used for detection of VE content in samples, such as animal/plant tissue, serum (plasma), drink, drug, oral liquid, food, etc.

Detection significance

VE is a kind of natural lipid-soluble antioxidant, which exist in cellular membrane structure (cell membrane, mitochondrial, microsomal membrane), lipid droplets of adipocytes and lipoproteins of plasma. It is scavenger of singlet oxygen and superoxide, blocking agent of lipid peroxidation and can protect protein sulfhydryl.

Detection principle

Fe3+ can be deoxidized to Fe2+ by VE with ferroin existing.Fe2+ can react with ferroin and form pink compound under certain condition. After colorimetric assay, VE content can be figured out according to the standard curve or calculated through formula.

Operation steps

1.  Detection of VE content in serum (plasma)

(1)  Extraction of n-heptane VE in serum (plasma) (n-heptane is not provided in this kit):

Vortex mixing (extract thoroughly) for 1 min, then centrifuge for 5~10 min at 3000~4000 rpm. Take the n-heptane VE extraction solution (the upper layer solution) for chromogenic reaction.

Note: Liquid in the tube will be divided into three layers, and the upper layer is n-heptane VE extraction solution, the middle layer is water and absolute ethyl alcohol and the lower layer is protein precipitate.

(2)  Chromogenic reaction:

Mix fully and stand for 2 min. Measure the OD values of each tube at 533 nm wavelength with cuvette of 1cm optical path, set to zero with absolute ethyl alcohol.

2.   Detection of VE content in tissue

(1)  Sample pretreatment:

Preparation of 10% homogenate solution: weigh the tissueaccurately. Add Reagent 4 (homogenate medium) in a weight (g): volume (mL) ratio of 1: 9, then use mechanical homogenization in ice water bath. Centrifuge the mixture solution at 2500 rpm for10 min and take the supernatant for measurement.

(2)  Extraction of n-heptane VE in tissue homogenate (n-heptane is not provided in this kit):

Vortex mixing (extract thoroughly) for 1 min, then centrifuge for 5~10 min at 3000~4000 rpm. Take the n-heptane VE extraction solution (the upper layer solution) for chromogenic reaction.

Note: Liquid in the tube will be divided into three layers, and the upper layer is n-heptane VE extraction solution, the middle layer is water and absolute ethyl alcohol and the lower layer is protein precipitate.

(3)  Chromogenic reaction:

       Mix fully and stand for 2 min. Measure the OD values of each tube at 533 nm wavelength with cuvette of 1cm diameter, set to 

       zero with absolute ethyl alcohol.