Extraction And Analysis Of Allantion In Department Of Wheat Crops

The allantoin in the crop of wheat lines are studied in the first time, exploring the extraction and purification conditions to determine the method of determination of allantoin and studying the capability of the hydroxyl radical scavenging, and the chiral enantiomers and the separation of isomers.Reflux and ultrasonic method are used in the single factor experiments and the orthogonal test, and the allantoin is extracted from flour and wheat bran flour of 7 crops and 2 commercial products to determine the optimum extracted condition is the reflux. Extraction reagents is 60% ethanol, solid to liquid ratio is 1:15, ultrasound is at 40℃, extraction time is 20 min, and extracted twice. The content is determined by HPLC, and on the hydroxyl radical scavenging and the enantiomers of chiral separation, concentration and clearance rate of hydroxyl radical as the stationary phase Kromasil C18 column, mobile phase is methanol: water (5:95, V:V); Flow rate is 0.5 mL / min; Column temperature is 35℃; Chiral to ovomucoid chiral stationary phase, mobile phase is methanol - potassium dihydrogen phosphate (0.01 mol / L) = 15:85 (V:V ), PH is 4.00; flow rate is 0.5 mL / min; column temperature is 25℃. The allantoin extract is purified by self-made column chromatography device, the eluent is chloroform: methanol (V:V) = 1:3, the flow rate is 2.0 mL / min, and the injection time is 30 seconds. Allantoin isomers is converted by the UV spectrophotometry.The coefficient of variation of allantoin content is determined less than 1.1% by HPLC; the recovery is 95.80% ~ 105.4%; the minimum detection limit is 0.5μg / mL; the concentration range of allantoin is 77.95 ~ 395.05μg / g, with descending order is Liaoning wheat bran, Liaoning wheat, barley bran, barley, Shandong wheat bran, Shandong wheat, barley bran, barley, oat bran, oats, buckwheat bran, buckwheat, Sichuan bran, Sichuan wheat, Hornsey flour and Loop flour. The capacity of hydroxyl free scavenging is proportional to the content of the base, the purity of purificationed allantoin is more than 80%, the allantoin enantiomer is fully separated by handling of splitting and separation is 2.750. When pH is 1.92 ~ 9.0, the allantoin all convert into ketone structure; When pH is 10.00 ~ 12.87, the allantoin all convert into enol by the UV detection.