Application

Detection significance

Plant total phenol is a common secondary natural metabolite in plants. There are several kinds of phenolic compounds, such as hydroxybenzoic acid, hydroxy cinnamic acid, flavonoids, chalcone, flavonoids, lignin, coumarin and astragalus. Phenolic compounds are antioxidants that delay or prevent oxidation and oxygen radical reactions.

Detection principle

Under alkaline conditions, tungsten-molybdenum acid can be reduced by phenols and produce blue compounds, which has a characteristic absorption peak at 760 nm. The content of total phenols in sample can be calculated indirectly by measuring the absorbance at 760 nm.

Experimental instrument

Incubator, 40 Mesh sieve, Ultrasonic cell disruptor, Centrifuge, Spectrophotometer (760 nm)

Pretreatment of sample

1.   Take fresh plant tissue (5-10 g), rinse the surface with distilled water and dry with filter paper. Then dry to constant weight in a vacuum drying oven at 40℃ (The difference between the two weights should be less 0. 3 mg). Crush and screen with 40 mesh sieve, sealed at room temperature.

2.    Weigh 0.1 g crushed sample and add 2.5 mL of 60% ethanol. Treat the sample with sonication (power: 300W, 3 seconds/time, interval for 4 seconds, total: 30 min). Centrifuge at 10000 g for 10 min at 25℃. Take the supernatant for detection.

Operation steps

1.    The dilution of standard

Dilute 1 mg/mL o-dihydroxybenzene solution with double distilled water to a serial concentration. The recommended dilution gradient is as follows: 150, 120, 100, 80, 40, 20, 0 μg/mL.

2.    Operation table

Technical parameter

1. The sensitivity of the kit is 0.73 μg/mL.

2. The intra-assay CV is 1.9% and the inter-assay CV is 2.5%.

3. The recovery of the kit is 101%.

4. The detection range of the kit is 0.73-150 μg/mL.

Notes

1. The kit is for scientific research only.

2. Instructions should be followed strictly, changes of operation may result in unreliable results.

3. The validity of kit is 6 months.

4. Do not use components from different batches of kit.

5. O-dihydroxybenzene standard solution should be prepared freshly, as it is easily oxidized.