Study on the bioactive ingredients of lamb abomasum medicine / 中华消化杂志

ABSTRACT

Objective:To study the material basis of the drug effect of the raw material of lamb′s tripe and vitamin B12 capsule.Methods:Rennet and pepsin were extracted with 0.9% sodium chloride and were purified by saturated ammonium sulfate precipitation, diethylaminoethyl-cellulose 52 and high pressure chromatography (HPLC) chromatography. Relative molecular weight was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) electrophoresis and liquid chromatography-triple quadrupole mass spectrometer. The amino acid composition and antioxidant activity of raw materials were analyzed with HPLC. The raw materials were completely extracted with water, phosphate buffer and sodium bicarbonate in turn, and in vitro 1, 1-diphenyl-2-picrylhydrazyl radil 2, 2-diphenyl-1-(2, 4, 6-trinitrophenyl)hydrazyl (DPPH) and 2, 2′-azinobis-(3-ethylbenzthiazoline-6-sulphonate) (ABTS) radical activity were measured. Glycoprotein from raw material was extracted with hot water and determined its growth-promoting activity gaginst Bifidobacterium adolescentis , Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis was measured. The composition of amino acid and monosaccharide were analyzed by HPLC and gas chromatography-mass spectrometry, respectively. Results:The enzyme activity of two purified rennet F6-2 and F7-2 which had different ionic strengths and pepsin F7-1 were 27 557.10, 17 532.60 and 17 728.15 U/g, respectively. The results of SDS-PAGE indicated that the molecular weights of three enzymes were similar, ranging from 35 000 to 40 000. The raw material contained 16 kinds of amino acids, of which hydrophobic amino acids accounted for 33.03% of the total amino acid content. When the sample concentration was 5 mg/mL, ABTS free radical scavenging activity of the three extracts was (37.80±0.45)%, (23.20±0.78)% and (62.80±0.74)%; DPPH free radical scavenging activity was (57.87±0.55)%, (5.03±0.25)% and (26.67±3.10)%, respectively. Glycoprotein extracts had promoted the growth of Bifidobacterium adolescentis, Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis, and there was statistically significant difference in the promotion of Lactobacillus delbrueckii subsp. Bulgaricus and Enterococcus faecalis ( P<0.05). The protein chain of glycoprotein was composed of 15 amino acids and the polysaccharide chain was composed of two monosaccharides, glucose and lactose. Conclusions:Two rennet and one pepsin are isolated and analyzed from raw material of lamb abomasum by various chromatographic methods. The raw material is rich in antioxidant active ingredients. The glycoprotein components of lamb abomasum has the activity of promoting the growth of probiotics.