A kind of calcium-containing compound separated from vinegar egg liquid and application thereof
A compound and composition technology, applied in the fields of medicine and food, can solve the problems of unclear functional components, unclear efficacy mechanism, no separation of calcium acetate compounds, etc., and achieve the effect of lowering blood pressure
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Embodiment 1
[0021] The extraction of embodiment 1 sugar calcium compound
[0022] (1) Preparation of Vinegar Egg Liquid: add 10kg eggs to 15kg 9° vinegar, soak for 24 hours, break the egg skin, soak for another 24 hours, filter to get the filtrate, and obtain Vinegar Egg Liquid.
[0023] (2) Concentrate 10L vinegar egg liquid to 2L volume, add 8L volume of 95% ethanol while stirring, let stand for more than 4h, filter to collect precipitate.
[0024] (3) Add 0.5 liters of water to dissolve the precipitate, filter to remove impurities, and separate the supernatant with an MCI column (5×150cm), first elute with 6 liters of deionized water, and then elute with 9 liters of 10% ethanol , the 10% ethanol eluate was collected and concentrated to a volume of 1.8 L.
[0025] (4) The 10% ethanol eluted concentrated solution is repeatedly separated by the ODS-AQ column, first eluted with deionized water, then eluted with 6% ethanol, and repeatedly separated by the ODS-AQ column for 2 to 3 times. T...
Embodiment 2
[0026] The identification of embodiment 2 sugar calcium compound
[0027] The compounds obtained in Example 1 were identified by HPLC\HPLC-MS and nuclear magnetic resonance methods, respectively.
[0028] (1)HPLC
[0029] Using ODSC18 chromatographic column, methanol: water = 6: 94 mobile phase, flow rate 0.3mL / min, detection wavelength 228nm chromatogram as shown figure 1 Shown; Retention time is that the chromatographic peak at 1.56min place is the compound that step 1 obtains.
[0030] (2)HPLC-MS
[0031] Using ESI+ mode, the voltage is 1800v, the mass spectrum is as follows figure 2 shown.
[0032] (3) NMR, with D 2 O is solvent, tetramethylsilane is internal standard, Bruck 500M Z NMR measurements were carried out.
[0033] Table 1 NMR results
[0034]
[0035] From 1 H-NMR shows δ5.11 (1H, d, J=3.6), 4.51 (1H, d, J=7.6), these two Hs should be the end group signal of the sugar, 3.0--4.1 is the H signal on the sugar, 1.79 (3H, s), should be a methyl signal...
Embodiment 3
[0037] The application of embodiment 3 sugar calcium compound in lowering blood pressure
[0038] In the present invention, the inhibitory effect on angiotensin-converting enzyme (ACE) is used to test the blood pressure-lowering effect of sugar and calcium. Prepare 100 μL of 50, 100, and 200 μg / mL sugar-calcium solutions, add 40 μL of 25 mU ACE enzyme solution respectively, incubate at 37 ° C for 30 minutes, add 200 μL of 1 mol / L hydrochloric acid, and measure the absorbance at a wavelength of 228 nm. Taking deionized water as the blank, the difference ratio between different concentrations of sugar calcium and the blank is the inhibition rate, and the results are shown in Table 2.
[0039] Table 2. Inhibitory effect of calcium sugar on ACE
[0040]
[0041] ACE is an angiotensin-converting enzyme, which acts through the renin-angiotensin system and the kallikrein-kinin system to increase blood pressure. Inhibiting the activity of ACE can lower blood pressure. It can be s...
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