First, the principle: Peroxidase is widely distributed in various tissues and organs of plants. In the presence of hydrogen peroxide, peroxidase can oxidize guaiacol to produce a brownish-brown material. This product has a maximum light absorption at 470 nm, so peroxide can be measured by measuring the change in absorbance at 470 nm. Enzyme activity.

Second, experimental materials: plant leaves

Third, equipment and reagents: electronic analytical balance, spectrophotometer, low-speed frozen multi-tube centrifuge, mortar, volumetric flask, measuring cylinder, test tube, straw.

(1) 100 mmol/L phosphate buffer (pH=6.0): [0.2 M Na2HPO 412.3 mL and NaH2PO 487.7 mL mixed and diluted 2 times]

(2) Reaction mixture: 50 ml of 100 mmol/L phosphate buffer (pH 6.0) in a beaker, 28 μl of guaiacol was added, and the mixture was heated and stirred on a magnetic stirrer until the guaiacol was dissolved, and after the solution was cooled, Add 19 μl of 30% hydrogen peroxide, mix well, and store in the refrigerator.

Fourth, the operation steps

1. Weigh 1g of plant material, put it into a mortar, grind it into powder with appropriate amount of liquid nitrogen, grind it into homogenate in the mortar with appropriate amount of phosphate buffer, centrifuge at 4000r/min for 15 minutes, transfer the supernatant to 100ml. In the volumetric flask, the residue was extracted once with 5 mL of phosphate buffer, and the supernatant was combined into a volumetric flask, and the volume was adjusted to the mark, and stored in a cold place for use.

2. Take 2 cuvettes with a light path of 1 cm, add 3 mL of the reaction mixture and 1 mL of phosphate buffer (or the enzyme solution boiled for 5 min) to one, and use the reaction mixture as the calibration control. 3ml, 1ml of the above enzyme solution (if the enzyme activity is too high to be diluted), immediately turn on the stopwatch recording time, measure the absorbance value at a wavelength of 470nm on a spectrophotometer, read once every 0.5min (30S), and read 3 times in succession. .

V. Calculation of results: A change of A470 of 0.01 per minute for 1 peroxidase activity unit (U)

Chinese name: guaiacol CP, >99.0% (GC)

English name: Guaiacol CP, >99.0% (GC)

Brand: Bo Fei Meike

Item No.: C16-G10224-100g

CAS: 90-05-1

Molecular formula: C7H8O2

Molecular weight: 124.14

Chemical properties: white or yellowish crystal or colorless to pale yellow transparent oily liquid. Has a special aromatic odor. Slightly soluble in water and benzene. Soluble in glycerin. Miscible with ethanol, ether, chloroform, oil, glacial acetic acid.

Uses: Used in pharmaceutical, dye and perfume intermediates. It is used to formulate flavors such as coffee, vanilla, smoked tobacco and tobacco. It is used in the synthesis of dyes and also as an analytical reagent. It is used in medicine to manufacture guaiacol sulfonate, and is used in the perfume industry to manufacture vanillin and artificial musk. Test copper, hydrocyanic acid and nitrite.

CANSPEC quality provides guaiacol, Guaiacol, CP, >99.0% (GC)|90-05-1 with excellent performance, and its excellent quality meets the diverse needs of your different experiments.

This product is provided by Shanghai Chuangsai quality, welcome to inquire!

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