| Shanxi Vinegar has a our long history, with its unique production process, become a necessary good for people’s daily life condiment. In addition, Shanxi Vinegar has polyphenols, a variety of functional ingredients vitamins, melanoidins, etc., they are given a unique Vinegar bactericidal anti-inflammatory, promote digestion, antioxidant and anti-aging, assisted lowering blood pressure and lowering blood sugar as well as lipid-lowering effect.These physiological activities have great significance to the development and innovative of vinegar’s functionality study. The reason of Vinegar for having these superior characteristics, mainly due to its unique technology smoked grains. Smoked Grains process is mainly occurred Maillard reaction, which can not only increase the colot and aroma of Vinegar which favorite by the public, but also played a significant role for improving the Vinegar’s health functions study.In this experiment, we chose the smoked grains of Shanxi Vinegar as the research object, through in the vitro bile micelles method to research the lipid-lowering activity of Vinegar smoked mash with different molecular weights. Contents and main conclusions of this paper are as follows:(1) In this study, the first we do is to chose the assay conditions of cholesterol by using high performance liquid chromatography (HPLC).In this part, we compare the affect of 100% methanol and 100% acetonitrile as two kinds of mobile phase,205nm and 208nm as two kinds of detection wavelengths to HPLC determination cholesterol levels peak shape, and finally the optimum chromatographic conditions for cholesterol determination by HPLC were determined:Column Diamonsil C18 (4.6×150, mm,5μm); mobile phase of 100% methanol; flow rate of 1.0mL/min; column greenhouse temperature; detection wavelength 205nm; injection volume 10μL. Under these conditions, the detection of resistance parallel, the cholesterol peak was maximum in the chromatogram picture, no impurity peaks, peak ideal, and the Stability of mobile phase is good. Using this method to chromatography each use liquid of cholesterol and draw the standard curve. Using the statistical software package SPSS 16.0 for Windows calculates the standard curve regression equation Y= 5E+06X+448177, correlation coefficient R2-0.9961.(2) Through the ultrafiltration technology, size exclusion chromatography and freeze-drying techniques,smoked unfiltered water extracts were separated to different molecular weight components and we obtain some smoked unfiltered water extract with different molecular weight, and then we measured the Lipid-lowering activity of them,aims to find out the best one. Ultrafiltration technology has been the first choice to separated the smoked unfiltered water extracts of different molecular components:less than 1KDa (W1),1-3 KDa (W2),3-5 KDa (W3),5-10 KDa (W4) and 10-100 KDa (W5) in five parts, freeze-dried, Lipid-lowering activity of each component are compared and screening. Analysis results find out the 3-5kDa components was highest Lipid-lowering activity part. The 3-5kDa components were separated to different molecular components by size exclusion chromatography, then we measured the Lipid-lowering activity of them and finally find we obtain:M1 (greater than lOKDa), M2 (5kDa-10 kDa), M3 (3kDa-5 kDa) and M4 (less than 3kDa) four parts, each part of the cholesterol-lowering activity of comparison and selection, its lipid-lowering activity were M3> M4> M2> M1, so the Lipid-lowering activity of 3-5kDa mainly because the contribution of M3.Determination the polyphenol content of the various parts from M1-M4 using Folin phenol method: descending sorting followed by M4, Ml, M2 and M3. I.e., the highest activity of polyphenol minimum hypolipidemic component content of the M3, only 17mg/mL. UV spectrophotometer spectrophotometric determination the Browning index of the W3 (3kDa-5 kDa) components,its browning index was 0.2085, the reaction to the content of melanoidins 780mg/g, explained M3 components was rich in melanoidins substances, made a great contribution for lipid-lowering activity.(3) By using the method of high strength sodium chloride dissociation melanoidins, the melanoidin 3-5kDa part further dissociated into two parts:The melanoidin skeleton (pure melanoidins PM) and small molecule connected to the melanoidin skeleton by non covalent bonds form quantity compound (BMC). Separately on the three part of PM, BMC and melanoidins 3-5kDa, analyzing the lipid-lowering activity and polyphenol content of them. The results showed, under the same concentration, the three parts’s lipid-lowering activity from high to low was that:PM>M33-skDa>BMC. However, the PM component of the polyphenol content was the lowest, only 22mg/g, and the polyphenol content of BMC was the highest, up to 50.7 mg/g. Description:The lowering blood lipid activity of melanoidin complexes (3KDa-5KDa) was mainly caused by the melanoidin core part of PM, and not related to the content of polyphenol and the small molecular BMC connected to the melanoidin skeleton.(4) By using in vitro bile micelles method to determination 100℃ steam smoked grains and traditional charcoal smoked grains 24h 1-5 days,two different techniques were compared with lipid-lowering activity.The results show that:After smoked grains inhibit cholesterol were higher than not smoked grains, indicating that smoked grains help to improve cholesterol-lowering activity; 4-5 days between new technology smoked grains cholesterol inhibition rate ranged from traditional crafts 24h; description of the new grains 24h craft smoked compared with the traditional process, to achieve the same effect in lowering the premise to shorten the time; The result suggest that based on the traditional steam smoked grains temperature(80-90℃),rasing the temperature in a reasonable range can not only greatly shorten the time of steam smoked grains processes,but also increase the Lipid-lowering activity of Shanxi Aged vinegar. |
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