Monascus yellow pigment compound as well as preparation method and application thereof
A technology for monascus yellow pigment and compound, which is applied in the field of monascus yellow pigment compound and its preparation, and can solve the problems of difficult quality control, blank design and processing, unclear composition and chemical composition, etc.
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Embodiment 1
[0038] Example 1: Separation and preparation of yellow pigment enriched fraction (MRG-MRI) in red yeast rice (monascus fungus fermentation product)
[0039] The strains of the Monascus fungus were inoculated into a liquid medium for activation to prepare a seed solution, and then the seed solution was inoculated into 100 bottles of 500mL Erlenmeyer flasks (70g of rice per bottle) for fermentation, first at 30°C Ferment for 3 days, then reduce the temperature to 25°C and ferment for 11 days to obtain the red yeast extract, then soak and extract with ethyl acetate three times to obtain the extract, and then concentrate under reduced pressure to obtain 202.3 g of ethyl acetate crude extract. Equal volume cyclohexane-methanol liquid phase extraction was performed on the crude ethyl acetate extract to obtain a cyclohexane layer (c, 56.2g) and a methanol layer (w, 100.0g). The total sample, cycloethane layer and methanol layer were analyzed by HPLC-DAD-ELSD, and it was found that th...
Embodiment 2
[0040] Example 2: Isolation and identification of monomeric compounds 1, 2, 3, 4, 5 in the yellow pigment-enriched fraction of red yeast rice (Monascus genus fungal fermentation product)
[0041] Using a preparative high-performance liquid chromatograph, the fraction MR H(3.5g) separation [chromatographic conditions: methanol-water-formic acid, 60:40:0.1, v / v / v, 8mL / min, Phenomenex Biphenyl column (21.2 × 250mm, 5μm)] to obtain 4 sub-fractions (MR H1 -MR H4 ), where MR H3 That is compound 3 (817.7mg, t R =32.0min); neutron fraction MR H2 (1.1g) was separated and obtained 9 sub-fractions (MR H21 -MR H29 ), the neutron fraction MR H23 Compound 2 (MR H231 ,3.0mg,t R = 47min); sub-fraction MR H22 (66.1 mg) was separated by HPLC semi-preparative [chromatographic conditions: acetonitrile-water-formic acid, 40:60:0.1, v / v / v, 3mL / min, Phenomenex Gemini C18 column (10×250mm, 5μm)] to obtain compound 1 (MR H221 ,5.0mg,t R = 45 min).
[0042] Using preparative high performan...
Embodiment 3
[0065] Embodiment 3: Stability evaluation of compounds 1-5
[0066] 3.1 Preparation of sample solution
[0067] Dissolve 1.0 mg of sample compounds 1, 2, 3, 4, and 5 in 20 μL DMSO to prepare a mother solution with a concentration of 50 mg / mL, then add water to dilute to 1 mg / mL to obtain a dilution, and put it in the refrigerator for later use.
[0068] Carry out the calculations in Table 3 below with the minimum sample volume:
[0069] table 3
[0070] sample Concentration (mg / mL) Volume (μL) solvent mother liquor 50 80 DMSO Diluent 1 4000 water+DMSO
[0071] 3.2 Analysis conditions of compounds
[0072] Using high performance liquid chromatography, different analysis conditions are used for liquid phase analysis of different compounds, and the chromatographic peaks are integrated to calculate the peak areas of the compounds at different sampling points at different temperatures or under different lighting conditions. Among them, the c...
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