Fermented product of coelogyne nitida and extract thereof

Abstract

The present application provides a fermenting product of white lily, which is obtained by fermenting a poria cocos fungus liquid. After the fermenting product of white lily provided by the present application is fermented by the poria cocos fungus, the ester groups in various lycoris tril group in the total lycoris tril are hydrolyzed, so that 4-hydroxy cinnamic acid and other carboxylic acids containing phenolic hydroxyl groups are obtained, which greatly improves the antioxidant property of the fermenting product of white lily. Meanwhile, due to the hydrolysis of lycoris tril, the bitterness of the fermenting product of white lily is reduced, and the taste of the fermenting product of white lily is improved. The present application also provides an extract of the fermenting product of white lily, which is obtained by crushing, sieving and then extracting the fermenting product of white lily with an organic solution.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of traditional Chinese medicine, in particular to Lilium lancifolium Thunb. fermentation product and extract thereof. BACKGROUND

[0002] Lilium is a traditional Chinese medicinal material, and is one of the first batch of medicinal and edible Chinese medicinal materials approved by the Ministry of Health. It is widely used in clinical practice of traditional Chinese medicine, and also has great research and development value as a raw material of food and health food. The Lilium medicinal material recorded in Chinese Pharmacopoeia (2020 edition) has three origins, which are the dry fleshy scales of Lilium lancifolium Thund., Lilium brownii F. E. Brown var. viriulum Baker or Lilium pumilum DC. The herbal textual research found that the taste and efficacy of Lilium recorded in different periods were quite different. For example, before the Ming Dynasty, the herbal record showed that Lilium had a sweet taste and was flat, and its function was to nourish lung qi and moisten lung dryness. However, since the Qing Dynasty, it has been considered that Lilium has a sweet and slightly bitter taste and is slightly cold, and its efficacy is gradually to clear lung heat. Lilium has a sweet taste and is good at nourishing lung qi and tonifying lung. However, Lilium lancifolium Thund. has a sweet and slightly bitter taste and is slightly cold, and its effect of clearing lung heat is more outstanding. The root cause is that cultivated Lilium has appeared since the Qing Dynasty. Compared with Lilium and Lilium pumilum DC., Lilium lancifolium Thund. has gradually become the mainstream variety of medicinal Lilium due to its wide adaptability.

[0003] Lilium lancifolium Thund. has a bitter taste, and the literature records that its quality is slightly inferior to that of Lilium due to the bitter taste. For example, the literature records show that “the one with bitterness is inferior” in the Ming Dynasty. The literature research found that Lilium lancifolium Thund. mainly contains saponins, polysaccharides, sterols, phenolic acid glycosides, flavonoids, phenylpropanoids, alkaloids and trace elements. Through literature research combined with transcriptome, metabolome, electronic tongue and molecular docking technology, it has been initially determined that phenolic acid glycosides (including Wangbailvy A, Wangbailvy B, Wangbailvy C, Wangbailvy D, Wangbailvy E, Wangbailvy F / Wangbailvy E, etc.) are the bitter taste components of Lilium, and the oxidation capacity of these components is weakened after being substituted. With the increasing market demand, the bitter taste of Lilium lancifolium Thund. not only affects the taste of food, but also affects the quality. Therefore, it is urgent to degrade these bitter taste components by related technologies or methods to improve the taste and antioxidant capacity of Lilium lancifolium Thund. SUMMARY

[0004] In order to solve the above technical problems, the present application provides a Lilium lancifolium Thund. fermentation product with low bitter taste components and high antioxidant capacity. Meanwhile, the present application also provides an extract of the Lilium lancifolium Thund. fermentation product.

[0005] To achieve the above-mentioned purposes, the present application adopts the following technical solutions:

[0006] A fermentation product of white lily, wherein the fermentation product of white lily is fermented by Poria cocos.

[0007] The fermentation comprises the steps of sterilizing and inoculating Poria cocos and culturing after soaking white lily in water.

[0008] The sterilization temperature is 120-130℃.

[0009] The sterilization time is 25-35min.

[0010] The inoculation amount of the Poria cocos liquid is 16-17g / L.

[0011] The absorbance value of the Poria cocos liquid is 0.8-1.0.

[0012] The culture temperature is 20-30℃.

[0013] The culture time is 28-35 days.

[0014] An extract of the fermentation product of white lily, wherein the extract is obtained by crushing, sieving and extracting the fermentation product of white lily with an organic solution.

[0015] The organic solution comprises methanol.

[0016] The content of total lycoriside in the extract of the fermentation product of white lily is less than 3.6mg / g. The content of total lycoriside is the sum of the contents of lycoriside A, lycoriside B, lycoriside C, lycoriside D, lycoriside E and lycoriside F.

[0017] The content of 4-hydroxycinnamic acid in the extract of the fermentation product of white lily is greater than 2.6mg / g.

[0018] The present application has the following beneficial effects relative to the prior art:

[0019] After the total lycoriside in the fermentation product of white lily is fermented by Poria cocos, the ester groups in various lycorisides in the total lycoriside are hydrolyzed, so that phenolic hydroxyl-containing carboxylic acids such as 4-hydroxycinnamic acid are obtained, which greatly improves the antioxidant property of the fermentation product of white lily. At the same time, due to the hydrolysis of lycoriside, the bitterness of the fermentation product of white lily is also reduced, and the taste of the fermentation product of white lily is improved. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 The relative content of bitter components in the fermentation product of white lily fermented by Poria cocos and Eurotium cristatum (golden flower fungus) for different fermentation times;

[0021] Figure 2 The extraction ion flow chart of the content change of lycoriside A-F in the fermentation process of white lily and Poria cocos.

[0022] Figure 3 Extraction ionogram of content change of liliin A-F in fermentation process of lycoris radiata and Eurotium cristatum;

[0023] Figure 4 Extraction ionogram of content change of 4-hydroxy cinnamic acid in fermentation process of lycoris radiata and Poria cocos;

[0024] Figure 5 Extraction ionogram of content change of 4-hydroxy cinnamic acid in fermentation process of lycoris radiata and Eurotium cristatum;

[0025] Figure 6 Antioxidant capacity chart of lycoris radiata fermented by Poria cocos and Eurotium cristatum with different fermentation time. DETAILED DESCRIPTION

[0026] In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present application. However, it will be apparent to one skilled in the art that the present application can be practiced without the specific details set forth in this description. In other instances, well-known methods have not been described in detail in order to avoid obscuring the present application.

[0027] Since lycoris radiata is a traditional Chinese medicine with bitter taste, its taste is poor, and the patient has poor compliance when taking it. The reason why lycoris radiata tastes bitter is that there is a high content of liliin in lycoris radiata. Specifically, there are at least six kinds of liliin in lycoris radiata, and the specific structures are as follows:

[0028] Liliin A

[0029] ;

[0030] Liliin B

[0031] ;

[0032] Liliin C

[0033] ;

[0034] Liliin D

[0035] ;

[0036] Liliin E

[0037] ;

[0038] Liliin F

[0039] .

[0040] The existence of the lycoruside makes the Cymbidium have strong bitter taste.

[0041] Based on this, the application provides a Cymbidium fermentation product, which is obtained by fermentation of Poria cocos.

[0042] The hydrolysis product (4-hydroxycinnamic acid) of lycoruside A, lycoruside B and lycoruside D:

[0043]

[0044] The hydrolysis product (caffeic acid) of lycoruside C and lycoruside E:

[0045]

[0046] The hydrolysis product (ferulic acid) of lycoruside F:

[0047]

[0048] The above hydrolysis product is the aglycone of lycoruside.

[0049] After fermentation, the bitter taste of Cymbidium is greatly reduced due to the hydrolysis of lycoruside in Cymbidium. At the same time, the aglycone is produced due to the hydrolysis, and the single electron pairing ability of the aglycone is stronger than that in 1,1-diphenyl-2-trinitrobenzene hydrazine (DPPH). Therefore, the production of aglycone improves the antioxidant property of Cymbidium fermentation product.

[0050] Specifically, the fermentation process needs to sterilize Cymbidium after soaking in water, then inoculate Poria cocos and culture Poria cocos. Specifically, the sterilization temperature is 120-130℃. Temperatures near this temperature can also be selected. The sterilization time is 25-35min. Then culture Cymbidium at 20-30℃ for 28-35 days to complete fermentation. The absorbance value (OD) of Poria cocos liquid used for inoculation is 0.8-1.0, and the amount of inoculated bacterial liquid is 500μL.

[0051] After crushing and sieving, the Cymbidium fermentation product can be extracted by an organic solvent to obtain the extract of Cymbidium fermentation product. Specifically, the organic solvent can be methanol. The total lycoruside content in the extract of Cymbidium fermentation product is less than 3.6mg / g, and the 4-hydroxycinnamic acid content in the extract of Cymbidium fermentation product is greater than 2.6mg / g. Therefore, the bitter taste of the Cymbidium fermentation product is reduced, and the antioxidant property is also improved.

[0052] The application is further described below in combination with specific examples.

[0053] ​​​Examples

[0054] 500 g of C. japonica were soaked in water, filtered and weighed. 18 g of the soaked C. japonica was placed in each plate and placed in a sterilized pot, 121 °C, 30 min, and then placed in a 25 °C incubator after cooling. The C. japonica was inoculated with E. turcicum liquid (1-20), P. igniarius liquid (21-40), and blank control (41-60) and grown for 30 days. Samples were taken every 5 days. The absorbance (OD) of the E. turcicum liquid and P. igniarius liquid used for inoculation was 0.8-1.0, and the amount of inoculated liquid was 500 μL.

[0055] The C. japonica-P. igniarius and C. japonica-E. turcicum fermentation samples were ground and passed through a No. 3 sieve. 0.5 g of each sample was precisely weighed into a 10 mL centrifuge tube, 6 mL of 75% methanol was precisely added, the weight was re-weighed, and ultrasonic extraction was performed for 40 min. The sample was cooled, the weight was re-weighed, and the lost weight was made up with 75% methanol. The sample was shaken and filtered through a 0.22 μm microporous filter. The filtrate was collected to obtain the sample solution. The blank solution was prepared by the same method. The sample solution and the blank were analyzed by HPLC-MS.

[0056] In some embodiments, the detection conditions of the high performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry technology are as follows:

[0057] Chromatographic conditions: C18 reverse phase chromatographic column; gradient elution; flow rate 1.0 mL / min; column temperature 30 °C; injection volume 10 μL; detection wavelength 210, 245, 306 nm; mobile phase: A, water containing 0.1% formic acid; B, methanol.

[0058] Mass spectrometry conditions: electrospray ionization (ESI); dry gas (N2) volume flow rate: 8.0 L / min; dry gas temperature: 320 °C; sheath gas temperature: 320 °C; nebulize 35 psig; sheath gas flow rate: 11 L / min; capillary voltage: 3.5 kV; nozzle voltage: 1.0 kV; collision voltage: 130 V; skimmer voltage 65 V; octopole RF 750 V; detection mode: negative ion detection; scan range: m / z 100-1500 Da.

[0059] By inputting the mass-to-charge ratio theoretical values of the bitter components of the bulb of fritillariae, namely, Wangbailvy A-F and their hydrolysis products in the mass spectrometry software, selecting "All" as the scanning mode, and setting the width range as ±0.02, the ion chromatogram can be extracted to extract the compounds. The compounds are integrated by automatic integration to obtain the peak area, and the relative content is obtained by dividing the peak area by 10000. The relative contents of the bitter components of the bulb of fritillariae, namely, Wangbailvy A-F and their hydrolysis products in the bulb of fritillariae of different fermentation dates are compared to determine the decrease of the bitter components.

[0060] Table 1. Contents of Wangbailvy A-F in the bulb of fritillariae samples (mg / g) (n=3)

[0061]

[0062] In the above table, FL represents Poria cocos, and JH represents Eurotium cristatum.

[0063] From Figure 1 , Figure 2 , Fig. 3 and Table 1, it can be seen that, in general, the content of the bitter components of the bulb of fritillariae is reduced after fermentation with Poria cocos and Eurotium cristatum, and especially, the contents of the bitter components Wangbailvy A-F are almost proportionally reduced after fermentation with Poria cocos. From Figure 1 , Figure 4 , Figure 5 and Table 2, it can be seen that the content of 4-hydroxy cinnamic acid is proportionally increased.

[0064] Table 2. Contents of 4-hydroxy cinnamic acid in the bulb of fritillariae during fermentation (mg / g) (n=3)

[0065]

[0066] The antioxidant capacity of the bulb of fritillariae fermented for different times was evaluated by the DPPH method. Specifically, the extract (1.2 mL) of the bulb of fritillariae fermented for different times was mixed with a DPPH solution (3.0 mL, 0.06 mg / mL). The mixture was vortexed for a few seconds and incubated in the dark at 37°C for 30 minutes. Then, the absorbance of the mixture was measured at 517 nm by a UV spectrophotometer. The blank control was prepared by adding 1.2 mL of the extract to 3.0 mL of the DPPH stock solution (0.06 mg / mL). The antioxidant activity was expressed as the percentage of DPPH radical elimination and calculated according to the following formula:

[0067] Scavenging activity (%) = [(Ablank-A sample) / Ablank] x 100%

[0068] Ablank and Asample are the absorbances of the blank control and the sample after adding DPPH, respectively. All tests were performed in triplicate.

[0069] FromFigure 6 It can be seen that with the continuous extension of fermentation time, the average DPPH scavenging activity of the C. morii fermentation product, i.e. the overall antioxidant capacity of the C. morii fermentation product, is improved. For example, after fermentation with T. fimbriatum for 30 days, the DPPH scavenging activity of C. morii is improved from 44.6% initially to 74.2%; after fermentation with A. penicillioides for 30 days, the DPPH scavenging activity of C. morii is improved from 34.0% initially to 52.1%.

Claims

1.A fermented product of Fritillaria pallidiflora, characterized in that: the fermented product of Fritillaria pallidiflora is obtained by fermentation of Fritillaria pallidiflora with Poria cocos liquid culture; the fermentation comprises the steps of sterilizing Fritillaria pallidiflora after soaking in water, inoculating Poria cocos liquid culture and culturing; the inoculation amount of the Poria cocos liquid culture is 16-17 g / L; the absorbance value of the Poria cocos liquid culture is 0.8-1.0; the temperature of the culturing is 20-30 ℃; the culturing time is 28-35 days. 2.The fermented product of Fritillaria pallidiflora according to claim 1, characterized in that: the sterilization temperature is 120-130 ℃. 3.The fermented product of Fritillaria pallidiflora according to claim 1, characterized in that: the sterilization time is 25-35 min. 4.An extract of the fermented product of Fritillaria pallidiflora, characterized in that: the extract is obtained by crushing, sieving and extracting with an organic solvent from the fermented product of Fritillaria pallidiflora according to claim 1; the organic solvent is methanol. 5.The extract of the fermented product of Fritillaria pallidiflora according to claim 4, characterized in that: the content of total veratrum glycosides in the extract of the fermented product of Fritillaria pallidiflora is less than 3.6 mg / g; the content of total veratrum glycosides is the sum of the contents of veratrum glycoside A, veratrum glycoside B, veratrum glycoside C, veratrum glycoside D, veratrum glycoside E and veratrum glycoside F. 6.The extract of the fermented product of Fritillaria pallidiflora according to claim 4, characterized in that: the content of 4-hydroxy cinnamic acid in the extract of the fermented product of Fritillaria pallidiflora is greater than 2.6 mg / g.

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