Reduction type coenzyme Q10 powder, composition thereof, and preparation method thereof

Abstract

The present invention describes a reduction type coenzyme Q10 powder, a composition thereof, and a preparation method thereof. The reduction type coenzyme Q10 powder is obtained by reacting an oxidation type coenzyme Q10 with the presence of a reducing agent, removing an organic solvent and other purities from a reaction solution after the reaction is finished to obtain an oil-soluble reduction type coenzyme Q10 liquid, and then directly performing prill formation with cold wind on an obtained reduction type coenzyme Q10 greasy substance. The obtained reduction type coenzyme Q10 powder has a lower crystallinity, and in a Cu-K[alpha] X-ray diffraction spectrum, has a strong peak at a diffraction angle 2[theta] being 18.9 DEG, and has a very strong absorption peak at a diffraction angle 2[theta] being 22.8 DEG. The reduction type coenzyme Q10 powder obtained in the present invention is in an incompletely crystallized state, has desirable stability and desirable oral bioavailability, and is suitable for use in applications such as dietary supplements, cosmetics or pharmaceuticals.

Claims

1. A method for preparing a reduced coenzyme Q10 powder, wherein the method comprises: (1) adding an oxidized coenzyme Q10 into n-hexane to form a reaction solution, heating the reaction solution to 50 C.60 C. temperature, adding a reductant for reacting with the heated reaction solution while, continuing to heat between 50 C.60 C., after the reaction is complete, creating a layer of n-hexane, and washing the n-hexane layer with 25 w/w % salt water and recycling the n-hexane at 35 C. in vacuum to obtain a molten and oily reduced coenzyme Q10 liquid, wherein the reductant is selected from thiosulfuric acid or sodium thiosulfate; and (2) heating the molten and oily reduced coenzyme Q10 liquid to 50 C., and then spraying the molten and oily reduced coenzyme Q10 liquid in a centrifugal sprayer or pressure sprayer, to form particles in cold air of not higher than 30 C. after spraying, to obtain a white reduced coenzyme Q10 powder.

2. The method according to claim 1, wherein the cold air is cold air with oxygen, cold air without oxygen, or cold nitrogen.

3. The method according to claim 1, further comprising adding an antioxidant, stabilizer, absorption enhancer and/or excipient as adjuvants to the reduced coenzyme Q10 liquid before spraying, to obtain the reduced coenzyme Q10 powder with adjuvants; or re-melting the reduced coenzyme Q10 powder, and then mixing the re-melted reduced coenzyme Q10 powder with an antioxidant, stabilizer, absorption enhancer and/or excipient as adjuvants, to obtain the reduced coenzyme Q10 powder by a spraying drying method.

4. The method according to claim 3, wherein the antioxidant is selected from one or more of vitamin E, butyl hydroxy toluene, ascorbic acid and vitamin A; the stabilizer is selected from one or more of gelatin, casein, sodium caseinate, soybean protein, cyclodextrin and gum Arabic; the absorption enhancer is selected from one or more of polyglycerol fatty acid ester, glycerin, stearic acid, oleic acid and linoleic acid; and the excipient is sugar and/or dextrin.

5. The method according to claim 1, wherein a dosage form of the reduced coenzyme Q10 powder is tablets or capsules, for dietary supplement.

Description

BRIEF DESCRIPTION OF FIGURES

(1) FIG. 1 is an X-ray diffraction pattern of the reduced coenzyme Q10 powder obtained by spraying oily reduced coenzyme Q10 in blowing cold air;

(2) FIG. 2 is an X-ray diffraction pattern of the reduced coenzyme Q10 powder obtained by crystallization in ethanol solvent of conventional methods;

(3) FIG. 3 is a DSC pattern of the reduced coenzyme Q10 powder obtained by spraying oily reduced coenzyme Q10 in blowing cold air;

(4) FIG. 4 is a DSC pattern of the reduced coenzyme Q10 powder obtained by crystallization in ethanol solvent of conventional methods.

DETAILED DESCRIPTION AND PREFERRED EMBODIMENTS THEREOF

(5) Hereafter, the present invention will be described specifically with reference to the examples. The examples are given only for illustration of the technical solution of the present invention and should not be construed to limit the present invention.

Example 1

(6) 100 g oxidized coenzyme Q10 is added to 1000 mL n-hexane, heated to 50 C. of temperature after mixing. 1000 mL 20% (w/w) sodium thiosulfate solution is added to the reaction solution and stirred at 50 C. for 1 hour, and then layered to a water layer as a lower layer and a n-hexane layer after finishing the reaction, the n-hexane layer is washed by 25% (w/w) salt water, and then recycle n-hexane at 35 C. in vacuum, to form a paste and then heat the paste to 50 C. of temperature, spray by centrifugal sprayer, and form pellet at 10 C. in cold air after spraying, to obtain white reduced coenzyme Q10 powder 96.8 g (Sample 1) with the purity is 99.5% and the average powder particle size is 154 mm.

(7) Experiment for Determining a Proportion of the Reduced Coenzyme Q10 in the Reduced Coenzyme Q10 Powder or Crystals by the HPLC Method

(8) Determining the proportion of reduced coenzyme Q10 in reduced coenzyme Q10 powder or crystals by the HPLC method. Determination conditions are as follows:

(9) Instruments: Agilient 1210

(10) Column: C18 column

(11) Mobile phase: ethanol/methanol=4/3(V/V)

(12) Detection wavelength: 210 nm

(13) Flow velocity: 1 ml/min

(14) The weight ratio of reduced coenzyme Q10 to oxidized coenzyme Q10 in Sample 1 by determination of the HPLC method is 99.2/0.8.

(15) It can be seen from it that the reduced coenzyme Q10 has a higher purity than 98% in the reduced coenzyme Q10 powder of the present invention.

(16) Experiment for Determining Diffraction Peaks of the Reduced Coenzyme Q10 Powder or Crystal by X-Ray Diffraction

(17) Determine the diffraction peak of the reduced coenzyme Q10 powder or crystal by X-ray diffraction method. It can determine from the X-ray diffraction pattern of diffraction peak position and intensity that the crystal shape changes. Determination of the present invention is made by a Cu-K ray X diffraction device. Determination conditions are as follows:

(18) Instrument models: X-ray powder diffraction Bruker D8 Advance

(19) Ray intensity: 40 kV 100 mA

(20) Angle range: 2=260

(21) Scanning speed: 2/min

(22) Scanning step: 0.05

(23) Divergence slit: 1

(24) Accept the slit: 0.60

(25) Scattering slit: 1

(26) X-ray diffraction pattern of Samples 1 determined by the Cu-K ray X diffraction method is shown in FIG. 1. The diffraction angle 2 in the diffraction pattern has a strong peak at 18.9, and has a very strong absorption peak at 22.8, and another diffraction angle peak intensity is decreased obviously, especially, a strength of peak 27.4 and 30.3 relative to peak 18.9 is less than 0.1. A peak strength at 22.8 is as 100. A peak strength at 20.0 is less than 40.0. A peak strength at 18.9 is less than 90.0.

(27) Experiment for Determining the Melting Point of the Reduced Coenzyme Q10 Powder or Crystal by Thermogravimetric Analyzer

(28) Determine the melting point of the reduced coenzyme Q10 powder or crystal by thermogravimetric analyzer. Degree of amorphous and crystallization of reduced coenzyme Q10 can be determined by the melting point. Determination conditions of the present invention are as follows:

(29) Instrument: Universal V4.7A TA Instruments

(30) Temperature scope: 30-100 C.

(31) Heating rate: 2.5 C./min

(32) Sample amount: 15 mg

(33) Samples 1 is scanned for melting point by differential scanning thermal analyzer, its differential scanning map is shown in FIG. 3. It can be seen from it that the melting point is 46.9 C.

(34) It can be seen from it that the crystal melting point of the present invention is lower than that of conventional solvent crystallization method, because the melting point of stable crystallization is higher, and the melting point of metastable crystallization is lower. The result illustrates that the crystallization degree of reduced coenzyme Q10 declines, its amorphous degree increases.

Example 2 (Comparative Example: Crystallization)

(35) 100 g oxidized coenzyme Q10 is added into 1000 mL n-hexane, heated to a 50 C. of temperature after mixing. 1000 mL 20% (w/w) sodium hydrosulfite solution is added to the reaction solution, and stirred at 50 C. for 1 hour and then. layered to a water layer as a lower layer and a n-hexane layer after finishing the reaction, the n-hexane layer is washed by 25% (w/w) salt water, and then recycle n-hexane at 35 C. in vacuum, to form a paste and then add 1100 ml of anhydrous ethanol and heated to 50 C. to be dissolved, then add 350 ml water to the solution under stirring, and cool to 5 C. to crystallization, to obtain 95.2 g white reduced coenzyme Q10 powder (sample Sample 2) after filtering and drying in vacuum, the purity is 99.4%, the average powder particle size is 148 mm.

(36) According to the method of Example 1, the proportion of reduced coenzyme Q10 of Sample 2 is determined by HPLC method, the weight ratio of reduced coenzyme Q10 to oxidized coenzyme Q10 in Sample 2 is 99.0/1.0.

(37) According to the method of Example 1, X-ray diffraction pattern of Samples 2 by the Cu-K ray X diffraction method is shown in FIG. 2. In the diffraction pattern, the diffraction Angle 2 has strong peak at 3.0, 4.6, 20.1, 22.8, 27.4, 30.3, the diffraction Angle 2 has a very strong absorption peak at 18.7, 18.9, and another diffraction Angle peak intensity is decreased obviously, the peak strength at peak 27.4 and 30.3 is less than 0.1 relative to peak 18.9. A peak strength at 22.8 is as 100. A peak strength at 20.0 is less than 40.0. A peak strength at 18.9 is less than 90.0.

(38) According to the method of Example 1, the melting point of Sample 2 is determined by thermogravimetric analyzer, Samples 2 by differential scanning thermal analyzer is scanned for melting point. The differential scanning map is shown in FIG. 4, its melting point is 49.55 C.

Example 3 (Comparative Example: Crystallization)

(39) 30 g Sample 2 of Example 2 is dissolved in 20 times of sunflower oil, to dissolve under stirring at 60 C. of temperature, and then cool to 5 C. to crystallization, remove vegetable oil on the crystal surface with n-hexane after filtering, and dry under reduced pressure to obtain 25.6 g reduced coenzyme Q10 (Sample 3) with the purity of 98.9%.

(40) According to the method of Example 1, the proportion of reduced coenzyme Q10 in Samples 3 is determined by HPLC method, the weight ratio of reduced coenzyme Q10 to oxidized coenzyme Q10 in Sample 2 is 99.1/0.9.

Example 4: Experiment Result of Stability

(41) Sample 1 and Sample 2 and Sample 3 obtained by Example 1, Example 2, Example 3 are respectively stored under shading or nitrogen for two months at 18 C. The retention rate of reduced coenzyme Q10 is respectively determined within 1 month and 2 months. The results are in Table 1.

(42) TABLE-US-00001 TABLE 1 The retention rate of reduced coenzyme The retention rate of reduced Sample Q10 within 1 month coenzyme Q10 within 2 months Sample 1 98.4% 95.5% Sample 2 75.4% 66.3% Sample 3 89.3% 85.4%

(43) It can be seen from the result of Table 1, the stability of the reduced coenzyme Q10 powder obtained by the method is the best one. It still reaches 95.5% after 2 months. It is higher than the reduced coenzyme Q10 obtained by conventional method, with levels of 66.3% of Samples 2, 85.4% of Sample 3.

Example 5: Experiment for Calculating Pharmacokinetic Parameters of Compounds and Relative Bioavailability Thereof

(44) Sample 1 and Sample 2 and Sample 3 obtained by Example 1, Example 2, Example 3 are used for beagle feeding experiment respectively, and the reduced coenzyme Q10 concentration in blood of feed beagle is determined, a basic pharmacokinetic parameters and relative bioavailability is calculated.

(45) Experimental scheme is as follows:

(46) 4 beagles, half male and half female, 9-12 kg weight, provided by Shanghai Institute of Medicine Center for experimental animals, laboratory animals use license: SYXK (Shanghai), 2010-0049. A cross double cycle test is made. Gastric gavage is respectively for Sample 1, 2, 3, to fill stomach in a dose of 20 mg/kg, a delivery volume of 2 ml/kg. Samples with a suspension of soybean oil mixture for medicine (filling water 20 ml after feeding), normally feeding and drinking before testing, feeding feed containing meat on morning of the same day of feeding the medicine, to fill the stomach after 30 min. Two cycle tests are interval for a week.

(47) Calculate the Pharmacokinetic Parameters of Compounds by WinNonlin 5.3 Software.

(48) Peak-reaching time Tmax and peak-reaching concentration Cmax use measured values. Drug concentration area under time curve AUC.sub.0-t value is calculated by the trapezoidal method, AUC.sub.0- is calculated by the formula AUC.sub.0-=AUC.sub.0-t+Ct/ke. Ct is a concentration measured by the last time point, ke is an eliminate rate constant, with half logarithmic mapping method, calculation by eliminating the concentration of the phase; Plasma elimination half-life t.sub.1/2=0.693/ke. Relative bioavailability F=(AUC.sub.0-t, Sample 2 or 3/AUC.sub.0-t, Sample 1)100%. The main pharmacokinetic parameters after Beagle lavage for 20 mg/kg sample. Refer to Table 2:

(49) TABLE-US-00002 TABLE 2 T.sub.max C.sub.max AUC.sub.0-t AUC.sub.0- MRT t.sub.1/2 Sample (h) (ng/ml) (ng .Math. h/ml) (ng .Math. h/ml) (h) (h) F (%) Sample 1 Average 7.5 303 3640 2865 10.9 6.11 / Standard 3.0 103 1833 2150 2.5 2.67 / deviation Sample 2 Average 4.5 261 2359 2405 7.77 3.94 61.9 Standard 1.0 78 1181 1200 0.36 0.60 52.1 deviation Sample 3 Average 6.3 287 2765 2658 8.35 4.76 74.7 Standard 1.9 83 1237 1890 1.81 1.76 49.5 deviation

(50) It can be seen from Table 2 that the bioavailability of reduced coenzyme Q10 powder (samples 1) obtained by the method in beagle is significantly higher than that of the other two ways.

(51) It can be seen from Examples 1 to 5 that in comparison with the crystallization method of the present invention and the Comparative Examples, the reduced coenzyme Q10 liquid of the present invention is sprayed in a sprayer, and particles after spraying in cold air has a lower crystallinity, and higher stability and bioavailability.

Example 6

(52) 120 mL acetone and 80 mL deionized water are added to 60 g oxidized coenzyme Q10, heated under stirring to 50 C., stirring for 1.0 hr after adding 60 g ascorbic acid sodium. 150 mL n-hexane is added and placed after finishing the reaction, remove water of the lower layer, n-hexane of the upper layer is washed with 25% brine, and then to obtain a paste after recovering n-hexane, heat the paste up to 50 C. 1 g of natural vitamin E and 0.5 g ascorbic acid as an antioxidant is added to a sprayer, and spray through the pressure sprayer, fog droplets is at 0 C. nitrogen flow and shape immediately, finally obtain white reduced coenzyme Q10 powder 59.5 g. The purity is 96.3%, the average powder particle size is 210 mm.

(53) According to the method of Example 1, the proportion of reduced coenzyme Q10 in the reduced coenzyme Q10 powder or crystals is determined by HPLC method, the proportion of reduced coenzyme Q10/oxidized coenzyme Q10 is 99.4/0.6.

(54) The reduced coenzyme Q10 powder is mixed with safflower seed oil, beeswax, and lecithin, to form a soft capsule, wherein the content of the reduced coenzyme Q10 is 50 mg per soft capsule for a dietary supplement.

Example 7

(55) 1 g vitamin A as antioxidant is added to 30 g reduced coenzyme Q10 powder obtained by Example 6, and then mixed with 100 g gelatin, 100 g sugar, 35 g dextrin, 25 g glycerin solution to a pressure sprayer for spraying, droplets after spraying is in hot air of 190 C. to dry and shape immediately, to obtain 284.5 g a white reduced coenzyme Q10 powder. The content of reduced coenzyme Q10 is 10.6%.

(56) According to the method of Example 1, the proportion of reduced coenzyme Q10 in the reduced coenzyme Q10 powder or crystals are determined by HPLC method, the proportion of reduced coenzyme Q10/oxidized coenzyme Q10 is 96.5/3.5.

Example 8

(57) 150 mL deionized water is added to 72 g oxidized coenzyme Q10, heated under stirring to 50 C., to add 35 g phosphorylation biological reductase for 4.0 hr and placed, to add 120 mL n-hexane to remove water of the lower layer, and wash out of n-hexane of the upper layer with 25% brine, to obtain paste after recovering n-hexane, the paste is heated up to 50 C., and then mixed with 0.5 g BHT, 0.5 g stearic acid, 3.0 g oleic acid, and 1.0 g linoleic acid in a pressure sprayer for spraying, droplets after spraying is in air without oxygen of 10 C. to dry and shape immediately, to obtain 60.5 g white reduced coenzyme Q10 powder, with a purity 45.9%, and an average powder particle size 192 mm.

Example 9

(58) 150 mL ethanol are added to 55 g oxidized coenzyme Q10, heated under stirring to 50 C., add 50 g carboxylic acid sodium disulfide under stirring for reaction 1.0 hr and placed after finishing the reaction, and add 120 mL n-hexane and 100 mL deionized water, remove water of the lower layer, and wash out of n-hexane of the upper layer with 25% brine, to obtain paste after recovering n-hexane, the paste is heated up to 50 C., and then mixed with 0.8 g antioxidant butylated hydroxy toluene (BHT) and 0.8 g vitamin A, 3.0 g polyglycerol fatty acid ester in a pressure sprayer for spraying, droplets after spraying is in nitrogen air of 30 C. to dry and shape immediately, to obtain white reduced coenzyme Q10 powder 53.5 g, with a purity of 90.9%, an average powder particle size of 203 mm.

Example 10

(59) 40 g reduced coenzyme Q10 powder of Example 9 is mixed with 25 g sodium caseinate, 18 g modified starch, 37 g cyclodextrin, 16 g gum Arabic, 10 g soybean protein solution in a centrifugal sprayer for spraying, droplets after spraying is in hot air of 180 C. to dry and shape immediately, to obtain white reduced coenzyme Q10 powder 142.5 g. The content of reduced coenzyme Q10 is 20.8%.

(60) The present invention is illustrated by the above examples, however, should understand that the present invention is not limited to special instance and implementation scheme described here. These special examples and implementation plans is aimed at helping the person skilled in the art to practice the present invention. The persons skilled in the art is easily from the spirit and scope of the present invention to further improve and perfect, so the present invention only restricts by the content and scope of the claims of the present invention, and its intention to cover all in the alternative solutions and equivalent solutions which included in appendix claim limit within the scope of the invention spirit.