COMPOSITION CONTAINING QUERCETAGETIN

Abstract

A composition contains quercetagetin and 6-hydroxykaempferol, and, optionally further includes patuletin. The composition has an excellent antioxidant effect, and can be used as an active additive in food, drugs, health care products, cosmetics, and feed. In addition, the composition can prompt animal growth, reduce ratio of feed to gain, and improve meat quality in the feed.

Claims

1. A composition containing quercetagetin, wherein the composition comprises quercetagetin and 6-hydroxykaempferol, and wherein a content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-43).

2. The composition according to claim 1, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-35).

3. The composition according to claim 1, wherein the composition further comprises patuletin, and a content ratio of quercetagetin:patuletin is 100:(1-40).

4. The composition according to claim 3, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-35); the content ratio is preferably selected from 100:(1-30) and 100:(1-25), and more preferably selected from 100:(1-20) and 100:(1-15); and the content ratio of quercetagetin:patuletin is selected from 100:(1-35).

5. A feed comprising the composition of claim 1.

6. A food comprising the composition of claim 1.

7. A grease composition comprising the composition of claim 1 and a grease.

8. A pigment composition comprising the composition of claim 1 and a pigment.

9. A vitamin composition comprising the composition of claim 1 and a vitamin.

10. A protein composition comprising the composition of claim 1 and a protein.

11. A carbohydrate composition comprising the composition of claim 1 and a carbohydrate.

12. (canceled)

13. The composition according to claim 1, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-30).

14. The composition according to claim 1, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-25).

15. The composition according to claim 1, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-20).

16. The composition according to claim 1, wherein the content ratio of quercetagetin:6-hydroxykaempferol is 100:(1-15).

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0032] FIG. 1 of the Drawings shows the HPLC chromatogram of the composition 9 prepared in Example 9;

[0033] FIG. 2 of the Drawings shows the HPLC chromatogram of the composition 10 prepared in Example 10;

[0034] FIG. 3 of the Drawings shows the HPLC chromatogram of the composition 11 prepared in Example 11;

[0035] FIG. 4 of the Drawings shows the HPLC chromatogram of the composition 12 prepared in Example 12;

[0036] FIG. 5 of the Drawings shows the HPLC chromatogram of the composition 13 prepared in Example 13;

[0037] FIG. 6 of the Drawings shows the color protection effect of composition 9 to composition 11 on lycopene, in which (a) is a blank control group, (b) is a control group after high temperature treatment, (c) is a sample containing composition 9 after high temperature treatment, (d) is a sample containing composition 10 after high temperature treatment, and (e) is a sample containing composition 11 after high temperature treatment.

SPECIFIC MODES FOR CARRYING OUT THE EMBODIMENTS

[0038] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be further explained in detail with examples below. It should be understood that the specific examples described here are only for explaining the present invention, but not intended for limiting the present invention.

I. Preparation of the Composition Containing Quercetagetin

[0039] The composition products of Examples 1-8 were prepared according to the raw materials and parts by weight in the following table.

TABLE-US-00002 Quercetagetin 6-Hydroxykaempferol Patuletin (parts by weight) (parts by weight) (parts by weight) Example 1 100 10 1 Example 2 100 11 3 Example 3 100 11 4 Example 4 100 12 4 Example 5 100 10 3 Example 6 100 7 3 Example 7 100 5 1 Example 8 100 5 0

II. The First Batch of Experiments

[0040] (1) Antioxidant Effect in Lipids

[0041] 5 parts of chicken oil and butter were taken respectively, each of which is 100 grams. Each 3 groups of chicken oil experimental groups and butter experimental groups were set up, and the composition of Example 6 was added according to the addition amount of 4, 6 and 8 ppm respectively; each 1 group of chicken oil control group and butter control group were set up, and ethoxyquin was added in an addition amount of 150 ppm; each 1 group of chicken oil blank group and butter blank group were set up, without adding any additives. The additives in each group were mixed evenly with the lipid, placed in a constant temperature incubator at 40° C., and samples were taken at 0, 3, 7, 14, 21 and 28 days to determine their peroxide value. The results show that the composition of Example 6 can significantly reduce the peroxide value of lipids and delay the process of lipid oxidation loss. The antioxidant effect is comparable to that of ethoxyquin added in an amount of 150 ppm.

TABLE-US-00003 TABLE 1 The influence of Example 6 on the peroxide value of chicken oil (mmol/g) Example 6 Ethoxyquin 0 ppm 4 ppm 6 ppm 8 ppm 150 ppm Day 0 1.144 ± 0.025  1.050 ± 0.076  1.039 ± 0.102  1.139 ± 0.075  1.083 ± 0.029  Day 3 1.272 ± 0.171a  1.111 ± 0.035ab  1.122 ± 0.098ab  1.100 ± 0.044ab 1.033 ± 0.067b Day 7 1.839 ± 0.084a 1.367 ± 0.087b 1.361 ± 0.050b 1.272 ± 0.039b 1.117 ± 0.088c Day 14 2.528 ± 0.096a 2.417 ± 0.000b 2.028 ± 0.192c 1.483 ± 0.067d 1.577 ± 0.092d Day 21 5.222 ± 0.542a 3.667 ± 0.726b 2.872 ± 0.042c 2.717 ± 0.044c 2.817 ± 0.224c Day 28 5.917 ± 1.013a 5.556 ± 0.268a 3.222 ± 0.141b 2.989 ± 0.251b 2.922 ± 0.325b

TABLE-US-00004 TABLE 2 The influence of Example 6 on the peroxide value of butter (mmol/g) Example 6 Ethoxyquin 0 ppm 4 ppm 6 ppm 8 ppm 150 ppm Day 0 1.572 ± 0.123  1.550 ± 0.088  1.606 ± 0.121  1.400 ± 0.169  1.411 ± 0.150  Day 3 1.978 ± 0.092a  1.522 ± 0.092bc 1.678 ± 0.190b  1.328 ± 0.092bc  1.444 ± 0.142bc Day 7 2.178 ± 0.125a 2.017 ± 0.132a 1.767 ± 0.029b 1.572 ± 0.269b 1.617 ± 0.104b Day 14 2.750 ± 0.083a 2.133 ± 0.088b 1.822 ± 0.211c 1.683 ± 0.196c 1.656 ± 0.075c Day 21 3.333 ± 0.083a 2.933 ± 0.161b 2.111 ± 0.127c 1.906 ± 0.092d 1.817 ± 0.100d Day 28 4.494 ± 0.243a 2.889 ± 0.337b 2.056 ± 0.173c 1.956 ± 0.075c 2.022 ± 0.197c

[0042] (2) Feeding of Broilers

[0043] Experimental groups I-VI, a total of six groups, were set up, the composition of Example 6 was added to the feed at 1.25, 2.5, 3.75, 5, 10, 20 ppm respectively, and one control group was set up, the feed used in the control group was the same as that of the experimental group except that the composition of Example 6 was not added. A total of 147 1-day-old healthy Ross broilers (male:female ratio=1:1, average weight 46.7±2.34 g) were randomly divided into 7 groups, with 21 chickens in each group, and were fed according to the above addition amount. The average daily feed intake, average daily weight gain and ratio of feed to gain of the broilers in the above 7 groups throughout the whole period (1 to 42 days of age) were studied. The results show that when the composition of Example 6 is added to the feed, the ratio of feed to gain can be reduced. At the same time, when the additive dosage is 3.75 ppm, the ratio of feed to gain is the lowest.

TABLE-US-00005 TABLE 3 Effect of the feeding of broilers Amount of the additives The ratio of Group Additives (ppm) feed to gain Normal — — 1.54 ± 0.10.sup.a control I the composition 1.25 1.51 ± 0.12.sup.a II of Example 6 2.5 1.52 ± 0.11.sup.a III 3.75 1.50 ± 0.12.sup.a IV 5 1.51 ± 0.15.sup.a V 10 1.53 ± 0.14.sup.a VI 20 1.54 ± 0.23.sup.a

III. The Second Batch of Experiments

Preparation Examples 9-17

[0044] Example 9: The flowers of marigold were extracted by acetone, then water and n-hexane phase were added to carry out liquid-liquid separation, the acetone aqueous phase was concentrated and filtered, and the filter cake was extracted with a mixed solvent of n-hexane and ethyl acetate at 1:0.5 to remove the impurities to obtain a composition containing quercetagetin, wherein the ratio of quercetagetin to 6-hydroxykaempferol is 100:3, which is composition 9.

[0045] Example 10: The French marigold were extracted by acetone, then water and n-hexane phase were added to carry out liquid-liquid separation, the acetone aqueous phase was concentrated and filtered, and the filter cake was extracted with a mixed solvent of n-hexane and ethyl acetate at 1:1 to remove the impurities to obtain a composition containing quercetagetin, wherein the ratio of quercetagetin to 6-hydroxykaempferol is 100:9, which is composition 10.

[0046] Example 11: The French marigold were extracted with n-hexane for degreasing, the obtained French marigold residues were extracted with methanol, concentrated and filtered, and the filter cake was extracted with ethyl acetate, the extract was concentrated and dried, and then repeatedly washed with pure water for 3 times to obtain a composition containing quercetagetin, wherein the ratio of quercetagetin to 6-hydroxykaempferol is 100:17, which is composition 11.

[0047] Example 12: The French marigold were extracted with n-hexane for degreasing, the obtained French marigold residues were extracted with ethyl acetate, the extract was concentrated and filtered, impurities were removed from the filter cake with n-hexane, and the solid phase was dried to obtain a composition containing quercetagetin, wherein the ratio of quercetagetin to 6-hydroxykaempferol is 100:22, and the ratio of quercetagetin to patuletin is 100:5, which is composition 12.

[0048] Example 13: The marigold were extracted with n-hexane for degreasing, the obtained marigold flower meal was extracted with 67% methanol, the extract was concentrated and filtered, the filter cake was extracted and purified with ethyl acetate, the extract was concentrated and dried, and then n-hexane was used to remove the impurities, and the solid phase was dried to obtain a composition containing quercetagetin, wherein the ratio of quercetagetin to 6-hydroxykaempferol is 100:31, and the ratio of quercetagetin to patuletin is 100:10, which is composition 13.

[0049] Example 14: The high-purity reference substances of quercetagetin and 6-hydroxykaempferol were mixed in proportion to obtain a composition with the ratio of quercetagetin to 6-hydroxykaempferol of 100:32, which is composition 14.

[0050] Example 15: The high-purity reference substances of quercetagetin and 6-hydroxykaempferol were mixed in proportion to obtain a composition with the ratio of quercetagetin to 6-hydroxykaempferol of 100:42, which is composition 15.

[0051] Example 16: The high-purity reference substances of quercetagetin, 6-hydroxykaempferol and patuletin were mixed in proportion to obtain a composition with the ratio of quercetagetin to 6-hydroxykaempferol of 100:10 and the ratio of quercetagetin to patuletin of 100:5, which is composition 16.

[0052] Example 17: The high-purity reference substances of quercetagetin, 6-hydroxykaempferol and patuletin were mixed in proportion to obtain a composition with the ratio of quercetagetin to 6-hydroxykaempferol of 100:10 and the ratio of quercetagetin to patuletin of 100:28, which is composition 17.

[0053] 1. Determination of Antioxidant Capacity In Vitro

[0054] Each of the composition of Examples 9-17, as well as single-component quercetagetin, 6-hydroxykaempferol, patuletin was taken, and the detection of DPPH and hydroxyl radical scavenging ability was performed. The specific test methods are as follows.

[0055] (1) Determination of DPPH Free Radical Scavenging Ability

[0056] Quercetin, 6-hydroxykaempferol, patuletin, and compositions 9-17 were used to prepare sample solutions with concentrations of 1, 2, 3, 4, 5, and 6 μg/ml in absolute ethanol. Then, DPPH was prepared into a 2×10.sup.−4 mol/L solution with absolute ethanol, and 2 mL of blank and sample solution were accurately absorbed and mixed with 2 mL of DPPH solution, respectively, the resultants were placed in a water bath at 30° C. in the dark for 30 min, and the absorbance A (blank) and A.sub.sample (sample) were measured at the wavelength of 517 nm.

[0057] Calculation Method for the Clearance:

[00001]$\mathrm{clearance}\%=\frac{A-A_{\mathrm{sample}}}{A}\times 100\%$

[0058] (2) Determination of Hydroxyl Radical Scavenging Ability (by Salicylic Acid Method)

[0059] Quercetin, 6-hydroxykaempferol, patuletin, and compositions 9-17 were used to prepare sample solutions with concentrations of 100, 150, 200, 250, 300 μg/ml in absolute ethanol. 9 mmol/L salicylic acid-ethanol solution, 9 mmol/L FeSO.sub.4 solution and 8.8 mmol/L H.sub.2O.sub.2 solution were prepared. Then, different samples were added according to the order in the following table, and placed in a water bath at 37° C. for 15 min, and the absorbance at 510 nm was measured. Calculation method:

[00002]$\mathrm{clearance}\%=\frac{A0-\left(\mathrm{Asample}-\mathrm{Abackground}\right)}{A0}\times 100\%$

TABLE-US-00006 Category A0 A.sub.sample A.sub.background Salicylic acid-ethanol solution 1 ml 1 ml 1 ml FeSO.sub.4 solution 1 ml 1 ml 1 ml Sample solution 1 ml 1 ml water 12 ml 11 ml 12 ml H.sub.2O.sub.2 solution 1 ml 1 ml

[0060] The results of the two tests are shown in Table 4 below:

TABLE-US-00007 TABLE 4 Half clearance for DPPH and hydroxyl radical Hydroxyl radical scavenging ability DPPH (IC50 μg/ml) (IC50 μg/ml) Quercetagetin 14.82 412 6-hydroxykaempferol 23.62 500 Patuletin 31.98 595 Composition 9 6.02 303 Composition 10 5.49 294 Composition 11 6.06 301 Composition 12 6.59 307 Composition 13 10.64 358 Composition 14 9.80 343 Composition 15 14.03 391 Composition 16 6.24 303 Composition 17 6.11 301

[0061] The results in the above table show that under the condition of achieving the same antioxidant effect, the amount of the composition in the present application used is lower than that of the single-component product.

[0062] 2. Antioxidant Effect in Fish Oil Powder

[0063] Fish oil powder containing quercetagetin, 6-hydroxykaempferol, patuletin or compositions 9-17 and blank fish oil powder were prepared according to the following ingredients and contents (based on active ingredients). After the antioxidant and fish oil powder were evenly mixed, the resultants were placed in an incubator at 40° C., and samples were taken in 28 days to measure the peroxide value. The results are shown in Table 5 below.

TABLE-US-00008 TABLE 5 Determination results of peroxide value in fish oil powder Peroxide value of Peroxide value of fish oil powder fish oil powder (mmol/g) with an (mmol/g) with an addition amount addition amount of 10 ppm of 100 ppm Blank 230 Quercetagetin 76.54 57.32 6-Hydroxykaempferol 142.14 121.24 Patuletin 120.46 108.57 Composition 9 7.64 7.12 Composition 10 6.17 5.54 Composition 11 10.45 11.23 Composition 12 14.32 13.78 Composition 13 37.56 40.34 Composition 14 38.26 41.54 Composition 15 70.38 53.54 Composition 16 12.13 11.14 Composition 17 11.88 12.46

[0064] The results in the above table show that, as an additive for lipid-based components, the amount of the composition of the present application used is lower than that of the single-component product under the condition of achieving the same antioxidant effect.

[0065] 3. Broiler Experiment

[0066] Composition 9 and composition 14 were added to the feed at 2, 6, 10 and 100 ppm (based on active ingredients) respectively as the experimental group, and a blank control group was set up. A total of 189 1-day-old healthy Ross broilers (half male and female, average weight 46.7±2.34 g) were randomly divided into 9 groups, with 21 chickens in each group, fed the same amount of the above-mentioned feeds, respectively, and the ratio of feed to gain of the above-mentioned 9 groups throughout the whole period (1-42 days of age) were studied. The results are shown in Table 6 below.

TABLE-US-00009 TABLE 6 Results of ratio of feed to gain of broilers Ratio of feed to gain Addition amount Blank 2.18 ± 0.08 Composition 9 2.11 ± 0.06 2 ppm Composition 14 2.21 ± 0.04 Composition 9 2.01 ± 0.07 6 ppm Composition 14 2.00 ± 0.08 Composition 9 2.02 ± 0.06 10 ppm Composition 14 2.04 ± 0.05 Composition 9 2.08 ± 0.09 100 ppm Composition 14 2.05 ± 0.05

[0067] The results in the above table show that the ratio of feed to gain can be reduced when composition 9 and composition 14 were added to the feed. At the same time, when the addition dosage is more than 6 ppm, the ratio of feed to gain is significantly lower than that of the blank control group.

[0068] 4. Color Protection Effect

[0069] The lycopene oleoresin and edible salt were thoroughly mixed and divided into four groups. Then, 200 ppm of compositions 9, 10 and 11 were added to three groups among them, and one group was used as a blank control group. The above four groups of samples were placed in an incubator at a constant temperature of 105° C. for 2 hours and then taken out, and their color changes were observed. The results are shown in FIG. 1 of the description.

[0070] The results show that compositions 9, 10 and 11 could all delay the loss of pigment at high temperature.

INDUSTRIAL APPLICABILITY

[0071] The present invention provides a composition containing quercetagetin, containing quercetagetin and 6-hydroxykaempferol, and further containing patuletin. The composition has an excellent antioxidant effect, and can be used as an active additive ingredient in food, drugs, health care products, cosmetics and feed. In addition, the composition can promote animal growth, reduce ratio of feed to gain, and improve meat quality in the feed.