The Study Of Enzymatic Method And Gas Chromatography For Determination Of Swainsonine

Oxytropis and Astragalus species known as Locoweeds are responsible for Locoism and other problems. Locoweeds are mainly distributed on the vast rangeland, exceeding 11 million hectares, in northwest, north and southwest China. As one of the toxins of locoweed, Swainsonine is important to the research of Locoism. As the deeply study of chemical composition and the control and utility of locoweed, it requires a large amount of swainsonine and Swainsonine quantitative detection of trace. Because of low efficiency, the former extraction methods are to be optimized. This experiment was developed the 40 KHz supersonic method and improve the extraction efficiency. The quantitative detection method of swainsonine became more practical and easier to manipulate after constituting the determination method of Swainsonine by internal standard gas chromatography and optimizingα-Mannosidase inhibition assay.1. The mixture of 1.0 kg Astragalus variabilis Bunge powder and water was disposed with 40KHz supersonic cleaner. After concentration and purification, the water solution was extracted with n-butanol and the bottom water extraction was disposed with n-butanol alkaline solution. Then extract it with dilute acid. Extract the concentrated acid liquid with ammoniated chloroform. Concentrate the ammoniated chloroform to residue. The residue was sublimed in 100℃olefin and 28 mg white crystal was collected. The crystal was preliminary determined as swainsonine. The extraction rate was 28 mg/kg.2. Swainsonine in Astragalus locoweed was detected by gas chromatography with me-Gal as internal standard, when oven temperature was 200℃, injection temperature was 300℃, detector(FID)temperature was 280℃and flow rate of carrier gas was 200 kPa. The analysis results of single-calibration-ratio was 1.067 8 and good linear relationship was obtained in the range of 0.047 to 6 mg/mL. The locoweed samples using ultrasonic assisted extraction and treatment of 732 cation exchange resin, through its analysis of the results found that supersonic can help with the extraction Swainsonine the extraction process less loss.3. Added 100μL citrate buffer solution, 20μL jake bean a-mannosodase, and 20μL sample solution per cell in turn. Then, 37℃incubated for 15 min; added 20μL p-nitrophenyl-a-D-mannopyranoside in each cell; 37℃incubated 90 min. Mensurated absorbance in 405 nm. SW concentration Y to their absorbance X (blank control - its average) regression analysis, a regression equation: Y = 0.0135X +0.1613, r = 0.9964, that SW sample volume at 0.05-1.25 mg / mL good linear relationship.4. After handled with supersonic-assisted extraction method, the swainsonine contents of different growth Period. Astragalus strictus Grah ex Benth and Oxytropis ochrocephala Bunge were determined by gas chromatography. The results showed that the content order was full Flowering Period > Pod-bearing Period > Budding Period> Branching Period.