Coix seed oil containing 16 glycerides, and pharmaceutical preparation and use thereof

Abstract

The present invention relates to Coix seed oil extracted from Semen Coicis, pharmaceutical preparations thereof, and the use thereof in the treatment of tumors. Specifically, the Coix seed oil contains 5 diglyceride and 11 triglyceride ingredients in the following mass percentages: 1,3-diolein 0.40-0.58%, 1-linolein-3-olein 0.91-1.31%, 1,2-diolein 0.24-0.35%, 1-olein-2-linolein 0.66-0.95%, 1,2-dilinolein 0.33-0.47%, trilinolein 4.87-6.99%, 1-olein-2,3-dilinolein 13.00-18.69%, 1-palmitin-2,3-dilinolein 5.25-7.54%, 1,3-diolein-2-linolein 13.23-19.02%, 1-palmitin-2-linolein-3-olein 10.26-14.75%, 1,3-dipalmitin-2-linolein 2.28-3.28%, triolein 14.44-20.76%, 1-palmitin-2,3-diolein 8.06-11.58%, 1-olein-2-linolein-3-stearin 1.37-1.97%, 1,3-dipalmitin-2-olein 1.52-2.19% and 1,2-diolein-3-stearin 1.29-1.86%.

Claims

1. A Coix seed oil, comprising 5 diglyceride and 11 triglyceride ingredients in the following mass percentages: 1,3-diolein 0.40-0.58%, 1-linolein-3-olein 0.91-1.31%, 1,2-diolein 0.24-0.35%, 1-olein-2-linolein 0.66-0.95%, 1,2-dilinolein 0.33-0.47%, trilinolein 4.87-6.99%, 1-olein-2,3-dilinolein 13.00-18.69%, 1-palmitin-2,3-dilinolein 5.25-7.54%, 1,3-diolein-2-linolein 13.23-19.02%, 1-palmitin-2-linolein-3-olein 10.26-14.75%, 1,3-dipalmitin-2-linolein 2.28-3.28%, triolein 14.44-20.76%, 1-palmitin-2,3-diolein 8.06-11.58%, 1-olein-2-linolein-3-stearin 1.37-1.97%, 1,3-dipalmitin-2-olein 1.52-2.19% and 1,2-diolein-3-stearin 1.29-1.86%.

2. The Coix seed oil of claim 1, having the following physicochemical constants based on the fatty oil detection specific gravity at 20 C. 0.916-0.920, refractive index at 20 C. 1.471-1.474, acid value <0.2, iodine value 100-106, saponification value 186-195; wherein said diglyceride and triglyceride ingredients are in the following mass percentages: 1,3-diolein 0.45-0.55%, 1-linolein-3-olein 1.03-1.25%, 1,2-diolein 0.27-0.33%, 1-olein-2-linolein 0.75-0.91%, 1,2-dilinolein 0.37-0.45%, trilinolein 5.47-6.69%, 1-olein-2,3-dilinolein 14.63-17.88%, 1-palmitin-2,3-dilinolein 5.90-7.21%, 1,3-diolein-2-linolein 14.88-18.19%, 1-palmitin-2-linolein-3-olein 11.55-14.11%, 1,3-dipalmitin-2-linolein 2.57-3.14%, triolein 16.25-19.86% 1-palmitin-2,3-diolein 9.07-11.08%, 1-olein-2-linolein-3-stearin 1.54-1.88%, 1,3-dipalmitin-2-olein 1.71-2.09% and 1,2-diolein-3-stearin 1.45-1.78%.

3. The Coix seed oil of claim 2, wherein said diglyceride and triglyceride ingredients are in the following mass percentages: 1,3-diolein 0.49-0.51%, 1-linolein-3-olein 1.12-1.16%, 1,2-diolein 0.29-0.31%, 1-olein-2-linolein 0.81-0.85%, 1,2-dilinolein 0.40-0.42%, trilinolein 5.96-6.20%, 1-olein-2,3-dilinolein 15.93-16.58%, 1-palmitin-2,3-dilinolein 6.43-6.69%, 1,3-diolein-2-linolein 16.20-16.87%, 1-palmitin-2-linolein-3-olein 12.57-13.09%, 1,3-dipalmitin-2-linolein 2.79-2.91%, triolein 17.69-18.42%, 1-palmitin-2,3-diolein 9.87-10.27%, 1-olein-2-linolein-3-stearin 1.68-1.74%, 1,3-dipalmitin-2-olein 1.86-1.94% and 1,2-diolein-3-stearin 1.58-1.65%.

4. The Coix seed oil of claim 1, obtained by a process comprising steps of: (1) Supercritical carbon dioxide extraction: Crushing Coix seeds into 20-100 mesh powder and extracting the powder using a supercritical CO.sub.2 extraction system in which Coix seed powder is put in 600 L2 extractors; the CO.sub.2 preheater, extractor and separation column are heated by jacketed circulating hot water to make the extraction temperature and separation temperature to be 33-45 C. and 30-45 C., respectively; the outlet temperatures of separator I and separator II are kept at 20-50 C. and 15-35 C., respectively; the liquid CO.sub.2 is pressed at a flow rate of 1-3t/h into the CO.sub.2 preheater via a high pressure pump, turning into a fluid in supercritical state; in the extractor, an oil is extracted into the CO.sub.2 fluid at a pressure of 19-23 Mpa; then the CO.sub.2 fluid with this oil enters the separation column in which the pressure is controlled to 7-10 Mpa to separate this oil; the CO.sub.2 gas out from the separation column enters sequentially into separator I and separator II in which the pressure is sustained at 5-7 Mpa and 4-6 Mpa, respectively; impurities such as water separated therefrom are discarded; the CO.sub.2 gas returns to liquid CO.sub.2 for reuse through a condenser; and a continuous extraction for 2-3 h affords a crude Coix seed oil; and (2) Refining process comprises steps of: adding petroleum ether (bp. 60 C.-90 C.) into the crude Coix seed oil obtained by the supercritical CO.sub.2 extraction in an amount of 65% of the oil weight; adding 2% NaOH aqueous solution in an amount ranging from 36% to 56% of the oil weight according to the acid value; after stirring the mixture for 10 min and standing for 18-24 h, removing the lower niger layer; washing the upper layer with purified water and letting stand for 18-24 h; after the removal of the lower waste water, washing the upper layer again; after another standing for 40-50 h, removing the lower waste water; and demulsifying the upper layer with acetone in an amount of 70%-90% of the oil weight; after standing for 2-4 h, removing the lower waste acetone and adding 3% to 8% of activated neutral alumina by weight of crude oil in the upper oil layer; stirring the mixture for 30 min, then filtering off the precipitation; heating the filtrate and adding 2% to 6% of activated mixed adsorbent by weight of crude oil; stirring the mixture for 30 min at 40-50 C. and then filtering off the precipitation; concentrating the filtrate under a reduced pressure to recover the solvent, then washing again with purified water; after standing for 1-2 h, removing the lower waste water and heating the upper oil layer and vacuum concentrating it under nitrogen atmosphere; then sterilizing the oil via dry heat sterilization under vacuum at 160-170 C. for 1-2 h; after cooling, filtering the oil through a 0.2 m microporous membrane; then split charging the obtained Coix seed oil in 500 mL glass infusion bottles, nitrogenizing and sealing the bottles.

5. The Coix seed oil of claim 4, wherein said refining process comprises steps of: adding petroleum ether (bp. 60 C.-90 C.) into the Coix seed oil obtained by the supercritical CO.sub.2 extraction in an amount of 65% of the oil weight; adding 2% NaOH aqueous solution in an amount ranging from 36% to 56% of the oil weight according to the acid value; after stirring the mixture for 10 min and standing for 20 h, removing the lower niger layer; washing the upper layer with purified water and letting standing for 22 h; after the removal of the lower waste water, washing the upper layer again; after standing for another 46 h, removing the lower waste water; demulsifying the upper layer with acetone in an amount of 70%-90% by weight of the crude oil and standing for 3 h; removing the lower waste acetone and adding 5% of activated neutral alumina by weight of crude oil in the upper oil layer; stirring the mixture for 30 min, then filtering off the precipitation; heating the filtrate, and adding 4% of mixed adsorbent of activated kaolin and activated carbon (1:1); stirring the mixture for 30 min at 40-50 C., and then filtering off the precipitation; concentrating the filtrate under a reduced pressure to recover the solvent, then washing again with purified water; after standing for 1 h, removing the lower waste water; heating the upper oil layer and vacuum concentrating it in nitrogen atmosphere; then sterilizing the concentrated oil via dry heat sterilization under vacuum at 160-170 C. for 2 h; after cooling, filtering the oil through a 0.2 m microporous membrane; then split charging the obtained Coix seed oil in 500 mL glass infusion bottles, nitrogenizing and sealing the bottles.

6. The Coix seed oil of claim 1, for use in the preparation of antitumor drugs for treatment of tumors selecting from a group consisting of lung cancer, liver cancer, pancreatic cancer, prostate cancer, ovarian cancer and breast cancer, in early, middle or late stage.

Description

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

Example 1 Preparation of Coix Seed Oil

(1) Supercritical carbon dioxide extraction: Coix seeds were crushed into 70 mesh powder and extracted using 600 L2 supercritical CO.sub.2 extractors. Coix seed powder was put in an extractor. The CO.sub.2 preheater, extractor and separation column were heated by jacketed circulating hot water, so that the extraction temperature and separation temperature reached 40 C. and 45 C., respectively, and the outlet temperatures of separator I and separator II were kept 50 C. and 35 C., respectively. Liquid CO.sub.2 was pressed into the CO.sub.2 preheater via a high pressure pump at a flow rate of 2 t/h, turning into a fluid in supercritical state. In the extractor, an oil was extracted into the CO.sub.2 fluid at a pressure of 20 Mpa. Then the CO.sub.2 fluid with this oil entered a separation column, and the pressure of the separation column was controlled to 7 Mpa to separate the oil. The CO.sub.2 gas out from the separation column entered sequentially into separator I and separator II, in which the pressure was sustained at 7 Mpa and 6 Mpa, respectively. Impurities like water separated therefrom were discarded. The CO.sub.2 gas returned to liquid CO.sub.2 for reuse through a condenser. A continuous extraction for 2.5 h afforded a crude Coix seed oil.

(2) Refining: To the crude Coix seed oil obtained by supercritical CO.sub.2 extraction was added 65% petroleum ether (60 C.) based on the oil weight. 45% NaOH aqueous solution (2%) based on the oil weight was added according to the acid value. After stirring for 10 min, then standing for 20 h, the lower niger layer was removed. The upper layer was washed with purified water and let stand for 22 h. After the removal of the lower waste water, the upper layer went on a second washing. After standing for another 46 h, the lower waste water was removed, and the upper layer was demulsified by adding 80% acetone based on the oil weight. After standing for 3 h, the lower waste acetone was removed. The upper oil layer was added 5% activated neutral alumina by weight of crude oil, stirred for 30 min, and filtered. The filtrate was heated, added with 4% activated mixed adsorbent of kaolin and carbon (1:1) by weight of the crude oil, stirred for 30 min at 45 C., and then filtered. The filtrate was concentrated under a reduced pressure to recover the solvent, and washed again with purified water. After standing for 1 h, the lower waste water was removed. The upper oil was concentrated, in nitrogen atmosphere, by heating under vacuum and then underwent dry heat sterilization by vacuum at 165 C. for 1.5 h. After cooling, the oil was filtered through a 0.2 m microporous membrane and split charged in 500 mL glass infusion bottles in nitrogen atmosphere, and the bottles were sealed. The Coix seed oil was thus obtained in a yield of 4.5%. Physicochemical constants were detected as: specific gravity at 20 C., 0.915; refractive index at 20 C., 1.471; acid value 0.18; iodine value 102; and saponification value 190.

Example 2 Preparation of Coix Seed Oil

(3) Supercritical carbon dioxide extraction: Coix seeds were crushed into 60 mesh powder and extracted using 600 L2 supercritical CO.sub.2 extractors. Coix seed powder was put in an extractor. The CO.sub.2 preheater, extractor and separation column were heated by jacketed circulating hot water, so that the extraction temperature and separation temperature reached 40 C. and 40 C., respectively, and the outlet temperatures of separator I and separator II were kept 20 C. and 15 C., respectively. Liquid CO.sub.2 was pressed into the CO.sub.2 preheater via a high pressure pump at a flow rate of 1 t/h, turning into a fluid in supercritical state. In the extractor, an oil was extracted into the CO.sub.2 fluid at a pressure of 22 Mpa. Then the CO.sub.2 fluid with this oil entered a separation column, and the pressure of the separation column was controlled to 8 Mpa to separate the oil. The CO.sub.2 gas out from the separation column entered sequentially into separator I and separator II, in which the pressure was sustained at 6 Mpa and 5 Mpa, respectively. Impurities like water separated therefrom were discarded. The CO.sub.2 gas returned to liquid CO.sub.2 for reuse through a condenser. A continuous extraction for 2 h afforded a crude Coix seed oil.

(4) Refining: To the crude Coix seed oil obtained by supercritical CO.sub.2 extraction was added 65% petroleum ether (90 C.) based on the oil weight. 56% NaOH aqueous solution (2%) based on the oil weight was added according to the acid value. After stirring for 10 min, then standing for 22 h, the lower niger layer was removed. The upper layer was washed with purified water and let stand for 20 h. After the removal of the lower waste water, the upper layer went on a second washing. After standing for another 48 h, the lower waste water was removed, and the upper layer was demulsified by adding 90% acetone based on the oil weight. After standing for 2 h, the lower waste acetone was removed. The upper oil layer was added 8% of activated neutral alumina by weight of crude oil, stirred for 30 min, and filtered. The filtrate was heated, added with 6% of activated mixed adsorbent by weight of the crude oil, stirred for 30 min at 42 C., and then filtered. The filtrate was concentrated under a reduced pressure to recover the solvent, and washed again with purified water. After standing for 2 h, the lower waste water was removed. The upper oil was concentrated, in nitrogen atmosphere, by heating under vacuum and then underwent dry heat sterilization by vacuum at 170 C. for 1.5 h. After cooling, the oil was filtered through a 0.2 m microporous membrane and split charged in 500 mL glass infusion bottles in nitrogen atmosphere, and the bottles were sealed. The Coix seed oil was thus obtained in a yield of 4.9%. Physicochemical constants were detected as: specific gravity at 20 C., 0.920; refractive index at 20 C., 1.473; acid value 0.19; iodine value 104; and saponification value 186.

Example 3 Preparation of Coix Seed Oil

(5) Supercritical carbon dioxide extraction: Coix seeds were crushed into 100 mesh powder and extracted using 600 L2 supercritical CO.sub.2 extractors. Coix seed powder was put in an extractor. The CO.sub.2 preheater, extractor and separation column were heated by jacketed circulating hot water, so that the extraction temperature and separation temperature reached 33 C. and 39 C., respectively, and the outlet temperatures of separator I and separator II were kept 30 C. and 20 C., respectively. Liquid CO.sub.2 was pressed into the CO.sub.2 preheater via a high pressure pump at a flow rate of 3 t/h, turning into a fluid in supercritical state. In the extractor, an oil was extracted into the CO.sub.2 fluid at a pressure of 19 Mpa. Then the CO.sub.2 fluid with this oil entered a separation column, and the pressure of the separation column was controlled to 9 Mpa to separate the oil. The CO.sub.2 gas out from the separation column entered sequentially into separator I and separator II, in which the pressure was sustained at 5 Mpa and 4 Mpa, respectively. Impurities like water separated therefrom were discarded. The CO.sub.2 gas returned to liquid CO.sub.2 for reuse through a condenser. A continuous extraction for 3 h afforded a crude Coix seed oil.

(6) Refining: To the crude Coix seed oil obtained by supercritical CO.sub.2 extraction was added 65% petroleum ether (80 C.) based on the oil weight. 36% NaOH (2%) aqueous solution based on the oil weight was added according to the acid value. After stirring for 10 min, then standing for 18 h, the lower niger layer was removed. The upper layer was washed with purified water and let stand for 18 h. After the removal of the lower waste water, the upper layer went on a second washing. After standing for another 42 h, the lower waste water was removed, and the upper layer was demulsified by adding 75% acetone based on the oil weight. After standing for 2 h, the lower waste acetone was removed. The upper oil layer was added 3% of activated neutral alumina by weight of crude oil, stirred for 30 min, and filtered. The filtrate was heated, added with 2% of activated mixed adsorbent (ibid) by weight of the crude oil, stirred for 30 min at 47 C., and then filtered. The filtrate was concentrated under a reduced pressure to recover the solvent, and washed again with purified water. After standing for 1 h, the lower waste water was removed. The upper oil layer was concentrated, in nitrogen atmosphere, by heating under vacuum and then underwent dry heat sterilization by vacuum at 160 C. for 2 h. After cooling, the oil was filtered through a 0.2 m microporous membrane and split charged in 500 mL glass infusion bottles in nitrogen atmosphere, and the bottles were sealed. The Coix seed oil was thus obtained in a yield of 4.7%. Physicochemical constants were detected as: specific gravity at 20 C., 0.918; refractive index at 20 C., 1.474; acid value 0.15; iodine value 100; and saponification value 194.

Example 4 Preparation of Coix Seed Oil

(7) Supercritical carbon dioxide extraction: Coix seeds were crushed into 30 mesh powder and extracted using 600 L2 supercritical CO.sub.2 extractors. Coix seed powder was put in an extractor. The CO.sub.2 preheater, extractor and separation column were heated by jacketed circulating hot water, so that the extraction temperature and separation temperature reached 35 C. and 42 C., respectively, and the outlet temperatures of separator I and separator II were kept at 40 C. and 30 C., respectively. Liquid CO.sub.2 was pressed into the CO.sub.2 preheater via a high pressure pump at a flow rate of 1.5 t/h, turning into a fluid in supercritical state. In the extractor, an oil was extracted into the CO.sub.2 fluid at a pressure of 21 Mpa. Then the CO.sub.2 fluid with this oil entered a separation column, and the pressure of the separation column was controlled to 10 Mpa to separate the oil. The CO.sub.2 gas out from the separation column entered sequentially into separator I and separator II, in which the pressure was sustained at 7 Mpa and 5 Mpa, respectively. Impurities like water separated therefrom were discarded. The CO.sub.2 gas returned to liquid CO.sub.2 for reuse through a condenser. A continuous extraction for 2 h afforded a crude Coix seed oil.

(8) Refining: To the crude Coix seed oil obtained by supercritical CO.sub.2 extraction was added 65% petroleum ether (70 C.) based on the oil weight. 50% NaOH (2%) aqueous solution based on the oil weight was added according to the acid value. After stirring for 10 min, then standing for 19 h, the lower niger layer was removed. The upper layer was washed with purified water and let stand for 21 h. After the removal of the lower waste water, the upper layer went on a second washing. After standing for another 50 h, the lower waste water was removed, and the upper layer was demulsified by adding 85% acetone based on the oil weight. After standing for 4 h, the lower waste acetone was removed. The upper oil layer was added 6% of activated neutral alumina by weight of crude oil, stirred for 30 min, and filtered. The filtrate was heated, added with 5% activated mixed adsorbent (ibid) by weight of the crude oil, stirred for 30 min at 50 C., and then filtered. The filtrate was concentrated under a reduced pressure to recover the solvent, and washed again with purified water. After standing for 1 h, the lower waste water was removed. The upper oil layer was concentrated, in nitrogen atmosphere, by heating under vacuum and then underwent dry heat sterilization by vacuum at 162 C. for 2 h. After cooling, the oil was filtered through a 0.2 m microporous membrane and split charged in 500 mL glass infusion bottles in nitrogen atmosphere, and the bottles were sealed. The Coix seed oil was thus obtained in a yield of 4.0%. Physicochemical constants were detected as: specific gravity at 20 C., 0.920; refractive index at 20 C., 1.471; acid value 0.16; iodine value 105; and saponification value 192.

Example 5 Preparation of Coix Seed Oil

(9) Supercritical carbon dioxide extraction: Coix seeds were crushed into 40 mesh powder and extracted using 600 L2 supercritical CO.sub.2 extractors. Coix seed powder was put in an extractor. The CO.sub.2 preheater, extractor and separation column were heated by jacketed circulating hot water, so that the extraction temperature and separation temperature reached 42 C. and 45 C., respectively, and the outlet temperatures of separator I and separator II were kept 35 C. and 25 C., respectively. Liquid CO.sub.2 was pressed into the CO.sub.2 preheater via a high pressure pump at a flow rate of 2.5 t/h, turning into a fluid in supercritical state. In the extractor, an oil was extracted into the CO.sub.2 fluid at a pressure of 23 Mpa. Then the CO.sub.2 fluid with this oil entered a separation column, and the pressure of the separation column was controlled to 8 Mpa to separate the oil. The CO.sub.2 gas out from the separation column entered sequentially into separator I and separator II, in which the pressure was sustained at 6 Mpa and 4 Mpa, respectively. Impurities like water separated therefrom were discarded. The CO.sub.2 gas returned to liquid CO.sub.2 for reuse through a condenser. A continuous extraction for 2.5 h afforded a crude Coix seed oil.

(10) Refining: To the crude Coix seed oil obtained by supercritical CO.sub.2 extraction was added 65% petroleum ether (80 C.) based on the oil weight. 40% NaOH (2%) aqueous solution based on the oil weight was added according to the acid value. After stirring for 10 min, then standing for 24 h, the lower niger layer was removed. The upper layer was washed with purified water and let stand for 24 h. After the removal of the lower waste water, the upper layer went on a second washing. After standing for another 44 h, the lower waste water was removed, and the upper layer was demulsified by adding 70% acetone based on the oil weight. After standing for 3 h, the lower waste acetone was removed. The upper oil layer was added 4% activated neutral alumina by weight of crude oil, stirred for 30 min, and filtered. The filtrate was heated, added with 3% activated mixed adsorbent (ibid) by weight of the crude oil, stirred for 30 min at 40 C., and then filtered. The filtrate was concentrated under a reduced pressure to recover the solvent, and washed again with purified water. After standing for 1.5 h, the lower waste water was removed. The upper oil layer was concentrated, in nitrogen atmosphere, by heating under vacuum and then underwent dry heat sterilization by vacuum at 165 C. for 1.5 h. After cooling, the oil was filtered through a 0.2 m microporous membrane and split charged in 500 mL glass infusion bottles in nitrogen atmosphere, and the bottles were sealed. The Coix seed oil was thus obtained in a yield of 4.3%. Physicochemical constants were detected as: specific gravity at 20 C., 0.916; refractive index at 20 C., 1.473; acid value 0.14; iodine value 101; and saponification value 192.

Example 6

(11) 8000 mg of Coix seed oil was dissolved in 10 ml n-hexane by using ultrasonic dissolving method, and prepared to be a Coix seed oil solution in acetone (50 mg/mL). This solution was separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP silica, 20*250 mm, 10 m; Mobile phase: n-hexane/acetone=94:6 (v/v); Injection volume 15 ml; Flow rate: 18 mL/min; ELSD Detector: temperature of drift tube 45 C., flow rate of carrier gas 2.0 L/min). Peak fraction at retention time of 15.8 min was collected and concentrated under vacuum at 30 C. The concentrated fraction was transferred into a 10 ml sample vial and blow-dried with nitrogen at ambient temperature to obtain a colourless oil, 1,3-diolein.

(12) Q-TOF/MS: quasi-molecular ion peaks [M+Na].sup.+=m/z 643.5277 (Calcd.=643.5272, C.sub.39H.sub.72O.sub.5Na), =4.

(13) .sup.1H-NMR data and .sup.13C-NMR data are shown in Table 3.

(14) TABLE-US-00003 TABLE 3 .sup.1H NMR and .sup.13C NMR data (CDCl.sub.3) Position .sup.1H NMR .sup.13C NMR C-1, 1 174.0 C-2, 2 2.33 (4H, t, J = 5.0 Hz) 34.3 C-3, 3 25.0 C-4, 4 29.3 C-5, 5 29.3 C-6, 6 29.3 C-7, 7 29.8 C-8, 8 27.3 C-9, 9 5.34 (2H, m) 129.9 C-10, 10 5.34 (2H, m) 130.2 C-11, 11 27.3 C-12, 12 29.9 C-13, 13 29.5 C-14, 14 29.7 C-15, 15 29.5 C-16, 16 32.1 C-17, 17 22.8 C-18, 18 0.87 (6H, t, J = 5 Hz) 14.3 C-1 4.19 (2H, dd, J = 11.6, 4.8 Hz) 65.2 4.13 (2H, dd, J = 11.6, 5.7 Hz) C-2 4.08 (1H, m) 68.6 C-3 4.19 (2H, dd, J = 11.6, 4.8 Hz) 65.2 4.13 (2H, dd, J = 11.6, 5.7 Hz)

Example 7

(15) 8000 mg of Coix seed oil was dissolved in 10 ml n-hexane by using ultrasonic dissolving method, and prepared to be a Coix seed oil solution in acetone (50 mg/mL). This solution was separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP silica, 20*250 mm, 10 m; Mobile phase: n-hexane/acetone=94:6 (v/v); Injection volume 15 ml; Flow rate: 18 mL/min; ELSD Detector: temperature of drift tube 45 C., flow rate of carrier gas 2.0 L/min). Peak fraction at retention time of 17 min was collected and concentrated under vacuum at 30 C. The concentrated fraction was transferred into a 10 ml sample vial and blow-dried with nitrogen at ambient temperature to obtain a colourless oil, 1-linolein-3-olein.

(16) Q-TOF/MS: quasi-molecular ion peaks [M+Na].sup.+=m/z 641.5121 (Calcd.=641.5115, C.sub.39H.sub.70O.sub.5Na), =5.

(17) .sup.1H-NMR data and .sup.13C-NMR data are shown in Table 4.

(18) TABLE-US-00004 TABLE 4 .sup.1H NMR and .sup.13C NMR data (CDCl.sub.3) Posi- .sup.13C Posi- .sup.13C tion .sup.1H NMR NMR tion .sup.1H NMR NMR C-1 174.8 C-1 174.8 C-2 2.35 (4H, t, J = 35.1 C-2 2.35 (4H, t, J = 35.1 7.6 Hz) 7.6 Hz) C-3 25.9 C-3 25.9 C-4 30.1 C-4 30.1 C-5 30.1 C-5 30.1 C-6 30.1 C-6 30.1 C-7 30.7 C-7 30.7 C-8 28.2 C-8 28.2 C-9 5.39 (1H, m) 131.0 C-9 5.39 (1H, m) 130.7 C-10 5.39 (1H, m) 129.1 C-10 5.39 (1H, m) 131.0 C-11 2.80 (2H, t, J = 26.6 C-11 28.2 6.6 Hz) C-12 5.39 (1H, m) 128.9 C-12 30.8 C-13 5.39 (1H, m) 131.2 C-13 30.3 C-14 28.2 C-14 30.6 C-15 30.5 C-15 30.3 C-16 32.5 C-16 32.9 C-17 23.6 C-17 23.7 C-18 0.91 (3H, t, J = 15.0 C-18 0.92 (3H, t, J = 15.1 5.0 Hz) 5.0 Hz) C-1 4.21 (2H, dd, J = 66.0 11.5, 4.3 Hz) 4.16 (2H, dd, J = 11.5, 5.7 Hz) C-2 4.11 (1H, m) 69.4 C-3 4.21 (2H, dd, J = 66.0 11.5, 4.3 Hz) 4.16 (2H, dd, J = 11.5, 5.7 Hz)

Example 8

(19) 8000 mg of Coix seed oil was dissolved in 10 ml n-hexane by using ultrasonic dissolving method, and prepared to be a Coix seed oil solution in acetone (50 mg/mL). This solution was separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP silica, 20*250 mm, 10 m; Mobile phase: n-hexane/acetone=94:6 (v/v); Injection volume 15 ml; Flow rate: 18 mL/min; ELSD Detector: temperature of drift tube 45 C., flow rate of carrier gas 2.0 L/min). Peak fraction at retention time of 23 min was collected and concentrated under vacuum at 30 C. The concentrated fraction was transferred into a 10 ml sample vial and blow-dried with nitrogen at ambient temperature to obtain a colourless oil, 1,2-diolein.

(20) Q-TOF/MS: quasi-molecular ion peaks [M+Na].sup.+=m/z 643.5277 (Calcd.=643.5272, C.sub.39H.sub.72O.sub.5Na), =4.

(21) .sup.1H-NMR data and .sup.13C-NMR data are shown in Table 5.

(22) TABLE-US-00005 TABLE 5 .sup.1H NMR and .sup.13C NMR data (CDCl.sub.3) Position .sup.1H NMR .sup.13C NMR C-1 173.9 C-1 173.5 C-2 2.33 (4H, t, J = 5.0 Hz) 34.2 C-2 34.4 C-3 25.0 C-3 25.1 C-4, 4 29.3 C-5, 5 29.3 C-6, 6 29.3 C-7, 7 29.8 C-8, 8 27.3 C-9, 9 5.35 (2H, m) 129.8 C-10, 10 5.35 (2H, m) 130.2 C-11, 11 27.3 C-12, 12 29.9 C-13, 13 29.5 C-14, 14 29.7 C-15, 15 29.5 C-16, 16 32.1 C-17, 17 22.7 C-18, 18 0.88 (6H, t, J = 5 Hz) 14.3 C-1 4.32 (2H, dd, J = 12.0, 4.6 Hz) 62.1 4.24 (2H, dd, J = 12.0, 5.6 Hz) C-2 5.08 (1H, m) 72.3 C-3 3.73 (2H, d, J = 3.2 Hz) 61.8

Example 9

(23) 8000 mg of Coix seed oil was dissolved in 10 ml n-hexane by using ultrasonic dissolving method, and prepared to be a Coix seed oil solution in acetone (50 mg/mL). This solution was separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP silica, 20*250 mm, 10 m; Mobile phase: n-hexane/acetone=94:6 (v/v); Injection volume 15 ml; Flow rate: 18 mL/min; ELSD Detector: temperature of drift tube 45 C., flow rate of carrier gas 2.0 L/min). Peak fraction at retention time of 24.5 min was collected and concentrated under vacuum at 30 C. The concentrated fraction was transferred into a 10 ml sample vial and blow-dried with nitrogen at ambient temperature to obtain a colourless oil, 1-olein-2-linolein.

(24) Q-TOF/MS: quasi-molecular ion peaks [M+Na].sup.+=m/z 641.5121 (Calcd.=641.5115, C.sub.39H.sub.70O.sub.5Na), =5.

(25) .sup.1H-NMR data and .sup.13C-NMR data are shown in Table 6.

(26) TABLE-US-00006 TABLE 6 .sup.1H NMR and .sup.13C NMR data (CDCl.sub.3) Posi- .sup.13C Posi- .sup.13C tion .sup.1H NMR NMR tion .sup.1H NMR NMR C-1 173.9 C-1 173.5 C-2 2.33 (2H, t, J = 34.2 C-2 2.33 (2H, t, J = 34.4 5.0 Hz) 5.0 Hz) C-3 25.0 C-3 25.1 C-4 29.3 C-4 29.3 C-5 29.3 C-5 29.5 C-6 29.3 C-6 29.3 C-7 29.8 C-7 29.9 C-8 27.3 C-8 27.4 C-9 5.37 (1H, m) 129.8 C-9 5.37 (1H, m) 130.2 C-10 5.37 (1H, m) 130.2 C-10 5.37 (1H, m) 128.2 C-11 25.8 C-11 2.77 (2H, t, J = 25.8 6.5 Hz) C-12 29.9 C-12 5.37 (1H, m) 128.0 C-13 29.5 C-13 5.37 (1H, m) 130.4 C-14 27.4 C-14 27.4 C-15 29.5 C-15 29.8 C-16 32.1 C-16 31.7 C-17 22.8 C-17 22.7 C-18 0.89 (3H, t, J = 14.3 C-18 0.88 (3H, t, J = 14.2 6.8 Hz) 6.8 Hz) C-1 4.32 (1H, dd, J = 62.1 11.9, 4.5 Hz) 4.23 (1H, dd, J = 11.9, 5.6 Hz) C-2 5.08 (1H, m) 72.3 C-3 3.73 (2H, d, J = 61.8 3.2 Hz)

Example 10

(27) 8000 mg of Coix seed oil was dissolved in 10 ml n-hexane by using ultrasonic dissolving method, and prepared to be a Coix seed oil solution in acetone (50 mg/mL). This solution was separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP silica, 20*250 mm, 10 m; Mobile phase: n-hexane/acetone=94:6 (v/v); Injection volume 15 ml; Flow rate: 18 mL/min; ELSD Detector: temperature of drift tube 45 C., flow rate of carrier gas 2.0 L/min). Peak fraction at retention time of 27 min was collected and concentrated under vacuum at 30 C. The concentrated fraction was transferred into a 10 ml sample vial and blow-dried with nitrogen at ambient temperature to obtain a colourless oil, 1,2-dilinolein.

(28) Q-TOF/MS: quasi-molecular ion peaks [M+Na].sup.+=m/z 639.4964 (Calcd.=639.4959, C.sub.39H.sub.68O.sub.5Na), =6.

(29) .sup.1H-NMR data and .sup.13C-NMR data are shown in Table 7.

(30) TABLE-US-00007 TABLE 7 .sup.1H NMR and .sup.13C NMR data (CDCl.sub.3) Posi- .sup.13C Posi- .sup.13C tion .sup.1H NMR NMR tion .sup.1H NMR NMR C-1 173.9 C-1 173.5 C-2 2.32 (4H, t, J = 34.2 C-2 2.35 (2H, t, J = 34.4 5.0 Hz) 5.0 Hz) C-3 25.0 C-3 25.1 C-4 29.3 C-4 29.3 C-5 29.5 C-5 29.5 C-6 29.3 C-6 29.3 C-7 29.9 C-7 29.9 C-8 27.4 C-8 27.4 C-9 5.37 (1H, m) 130.2 C-9 5.37 (1H, m) 130.2 C-10 5.37 (1H, m) 128.2 C-10 5.37 (1H, m) 128.2 C-11 2.77 (4H, t, J = 25.8 C-11 2.77 (2H, t, J = 25.8 6.5 Hz) 6.5 Hz) C-12 5.37 (1H, m) 128.0 C-12 5.37 (1H, m) 128.0 C-13 5.37 (1H, m) 130.4 C-13 5.37 (1H, m) 130.4 C-14 27.4 C-14 27.4 C-15 29.8 C-15 29.8 C-16 31.7 C-16 31.7 C-17 22.7 C-17 22.7 C-18 0.89 (3H, t, J = 14.2 C-18 0.89 (3H, t, J = 14.2 6.8 Hz) 6.8 Hz) C-1 4.32 (1H, dd, J = 62.1 11.9, 4.6 Hz) 4.24 (1H, dd, J = 12.0, 5.6 Hz) C-2 5.08 (1H, m) 72.3 C-3 3.73 (2H, d, J = 61.8 3.2 Hz)

Example 11 Isolation and Identification of Trilinolein

(31) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C.sub.18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 12.6-14.2 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give the trilinolein.

(32) HR-EI-MS: m/z=878.7344 (Calcd.=878.7363, C.sub.57H.sub.98O.sub.6), Degree of unsaturation=9.

(33) IR (KBr film): 1746, 1170, 1098; 2928, 2856, 724; 3008, 1655 cm.sup.1 (weak).

(34) .sup.1H-NMR data are shown in Table 8.

(35) .sup.13C-NMR data are shown in Table 9.

(36) TABLE-US-00008 TABLE 8 .sup.1H-NMR spectral data (CDCl.sub.3) of the compounds of Examples 6-13 No. G-H H 2-H 3-H 4-H 5-H 6-H 7-H 8-H 9-H 10-H 11-H 12-H 13-H 14-H 15-H 16-H 17-H 18-H A 4.30 LLL 5.27 2.32 1.61 1.32 2.05 5.36 2.77 5.36 2.05 1.32 0.89 4.15 B 4.29 2.77 5.37 2.04 OLL 5.27 2.32 1.61 1.33 2.04 5.37 1.33 0.88 4.14 2.04 1.33 C 4.30 2.05 5.36 2.77 5.36 2.05 1.31 0.88 PLL 5.27 2.31 1.61 1.31 4.15 1.31 1.31 0.88 D 4.30 2.05 1.32 OLO 5.27 2.32 1.61 1.32 2.05 5.36 1.32 0.89 4.15 2.77 5.36 2.05 E 4.15 2.04 5.35 2.04 1.28 1.28 0.88 PLO 5.27 2.31 1.61 1.28 2.77 5.35 2.04 1.28 4.30 1.28 0.88 F 4.15 1.28 0.88 PLP 5.27 2.31 1.61 1.28 2.05 5.36 2.77 5.36 2.05 1.28 0.88 4.30 1.28 0.88 G 4.15 OOO 5.27 2.31 1.61 1.28 2.00 5.34 2.00 1.28 0.88 4.30 H 4.15 2.04 5.34 2.04 1.27 0.88 POO 5.27 2.31 1.61 1.28 4.30 1.27 0.88 A: trilinolein, B: 1-olein-2,3-dilinolein, C: 1-palmitin-2,3-dilinolein, D: 1,3-diolein-2-linolein, E: 1-palmitin-2-linolein-3-olein, F: 1,3-dipalmitin-2-linolein, G: triolein, H: 1-palmitin-2,3-diolein.

(37) TABLE-US-00009 TABLE 9 .sup.13C-NMR spectral data (CDCl.sub.3) of the compounds of Examples 6-13 No. Abb. ClC C-1 C-2 C-3 C-4 C-5 C-6 C-7 C-8 C-9 C-10 A 62.12 173.28 34.05 24.86 29.05~29.62 27.21 130.01 128.08 LLL 68.91 172.87 34.21 24.90 129.98 128.09 B 62.12 173.28 34.04 24.86 29.07~29.79 27.22 130.03 128.03 OLL 68.89 172.87 34.21 24.90 27.19 130.00 128.10 173.29 129.73 130.03 C 62.12 173.30 34.04 24.89 29.06~29.72 27.21 130.02 128.08 PLL 68.90 172.88 34.20 24.85 129.99 128.09 173.34 34.07 24.38 29.06~29.72 D 62.12 173.29 34.05 24.86 29.07~29.79 27.20 129.73 130.03 OLO 68.89 172.87 34.21 24.90 27.22 130.00 128.10 E 62.11 173.28 34.04 24.85 29.06~29.78 27.18 129.71 130.01 PLO 63.91 172.86 34.20 24.87 27.21 129.98 128.09 173.32 34.06 24.98 29.06~29.78 F 62.09 173.32 34.05 24.86 29.05~29.70 29.05~29.70 PLP 63.89 172.86 34.19 24.88 27.20 129.97 128.08 G 62.12 173.29 34.04 24.86 29.07~9.78 27.19 129.72 130.02 OOO 68.90 172.87 34.21 24.90 129.69 130.03 H 62.12 173.31 34.04 24.88 29.06~29.78 27.19 129.72 130.02 POO 68.90 172.90 34.21 24.86 129.69 130.03 173.35 34.06 24.90 29.06~29.78 No. Abb. C-11 C-12 C-13 C-14 C-15 C-16 C-17 C-18 A 25.64 127.91 130.22 27.21 29.05~29.62 31.53 22.58 14.07 LLL 127.90 B 25.65 127.91 130.24 27.24 29.07~29.79 31.55 22.60 14.10 OLL 127.90 27.22 29.07~29.79 31.93 22.71 14.14 C 25.64 127.909 130.236 29.06~29.72 31.54 22.59 14.09 PLL 127.898 130.236 31.95 22.71 14.14 D 27.24 29.07~29.79 31.93 22.71 14.14 OLO 25.65 127.90 130.24 27.24 29.07~29.79 31.55 22.60 14.10 E 27.21 29.06~29.78 31.92 22.69 14.12 PLO 25.64 127.90 130.22 27.23 29.06~29.78 31.54 22.58 14.07 31.94 22.71 14.12 F 29.05~29.70 31.93 22.69 14.12 PLP 25.63 127.89 130.22 27.20 29.05~29.70 31.53 22.58 14.07 G 27.24 29.07~29.78 31.92 22.70 14.12 OOO H 27.24 29.06~29.78 31.92 POO 22.70 14.12 29.06~29.78 31.94 22.70 14.12 A: trilinolein, B: 1-olein-2,3-dilinolein, C: 1-palmitin-2,3-dilinolein, D: 1,3-diolein-2-linolein, E: 1-palmitin-2-linolein-3-olein, F: 1,3-dipalmitin-2-linolein, G: triolein, H: 1-palmitin-2,3-diolein.

Example 12 Isolation and Identification of 1-Olein-2,3-Dilinolein

(38) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 15.4-17.3 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1-olein-2,3-dilinolein.

(39) HR-EI-MS: m/z=880.7518 (Calcd.=854.7363, C.sub.55H.sub.98O.sub.6), Degree of unsaturation=7.

(40) IR (KBr film): 1747, 1164, 1098; 2925, 2854, 723; 3008, 1655 cm.sup.1 (weak).

(41) .sup.1H-NMR data are shown in Table 8.

(42) .sup.13C-NMR data are shown in Table 9.

Example 13 Isolation and Identification of 1-Palmitin-2,3-Dilinolein

(43) Preliminary isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). A solution of Coix seed oil (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 17.4-18.1 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen, to give a crude product.

(44) The second purification was proceeded on Superstar Benetnach C.sub.18 Column (10 mm250 mm, 5 m) with mobile phase A: acetonitrile and mobile phase B: acetonitrile/tetrahydrofuran (1:1). A solution of the above crude product (20 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-23 min: 50%-60%, 32-43 min: 60%-90%, 43-60 min: 100%; and flow rate: 3 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 31.2-34.7 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1-palmitin-2,3-dilinolein.

(45) HR-EI-MS: m/z=854.7370 (Calcd.=854.7363, C.sub.55H.sub.98O.sub.6), Degree of unsaturation=7.

(46) IR (KBr Film): 1746, 1165, 1095; 2926, 2854, 722; 3009, 1648 cm.sup.1 (weak).

(47) .sup.1H-NMR data are shown in Table 8.

(48) .sup.13C-NMR data are shown in Table 9.

Example 14 Isolation and Identification of 1,3-Diolein-2-Linolein

(49) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 18.4-20.2 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1-olein-2,3-dilinolein.

(50) HR-EI-MS: m/z=882.7678 (Calcd.=882.7672, C.sub.57H.sub.102O.sub.6), Degree of unsaturation=7.

(51) IR (KBr film): 1747, 1163, 1097; 2925, 2855, 723; 3007, 1655 cm.sup.1 (weak).

(52) .sup.1H-NMR data are shown in Table 8.

(53) .sup.13C-NMR data are shown in Table 9.

Example 15 Isolation and Identification of 1-Palmitin-2-Linolein-3-Olein

(54) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 20.3-21.4 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1-palmitin-2-linolein-3-olein.

(55) HR-EI-MS: m/z=856.7519 (Calcd.=856.7513, C.sub.55H.sub.100O.sub.6), Degree of unsaturation=6.

(56) IR (KBr film): 1747, 1164, 1098; 2925, 2854, 723; 3008, 1655 cm.sup.1 (weak).

(57) .sup.1H-NMR data are shown in Table 8.

(58) .sup.13C-NMR data are shown in Table 9.

Example 16 Isolation and Identification of 1,3-Dipalmitin-2-Linolein

(59) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 25.7-26.2 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1,3-dipalmitin-2-linolein.

(60) HR-EI-MS: m/z=830.7371 (Calcd.=830.7363, C.sub.53H.sub.98O.sub.6), Degree of unsaturation=5.

(61) IR (KBr film): 1747, 1164, 1098; 2925, 2854, 723; 3008, 1655 cm.sup.1 (weak).

(62) .sup.1H-NMR data are shown in Table 8.

(63) .sup.13C-NMR data are shown in Table 9.

Example 17 Isolation and Identification of Triolein

(64) Isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile, Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). Coix seed oil solution (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 26.6-27.7 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give triolein.

(65) HR-EI-MS: m/z=884.7851 (Calcd.=884.7833, C.sub.57H.sub.104O.sub.6), Degree of unsaturation=6.

(66) IR (KBr film): 1749, 1165, 1095; 2925, 2854, 723; 3004, 1654 cm.sup.1 (weak).

(67) .sup.1H-NMR data are shown in Table 8.

(68) .sup.13C-NMR data are shown in Table 9.

Example 18 Isolation and Identification of 1-Palmitin-2,3-Diolein

(69) Preliminary isolation was carried out on P3000A preparative high performance liquid chromatography (Column: Superstar Benetnach C18, 20 mm150 mm, 5 m; Mobile phase A: acetonitrile; Mobile phase B: acetonitrile/tetrahydrofuran (1:1)). A solution of Coix seed oil (50 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-27 min: 50%-60%, 27-35 min: 90%, 35-45 min: 100%; and flow rate: 18 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 28.2-29.3 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen, to give crude product.

(70) The second purification was proceeded on Superstar Benetnach C.sub.18 Column (10 mm250 mm, 5 m) with mobile phase A: acetonitrile and mobile phase B: acetonitrile/tetrahydrofuran (1:1). A solution of the above crude product (20 mg/mL) was prepared with mobile phase B, and the injection volume for each separation was 1.5 mL. Gradient conditions were: mobile phase B: 0-23 min: 50%-60%, 32-43 min: 60%-90%, 43-60 min: 100%; and flow rate: 3 mL/min. UV detection was conducted at 208 nm. Peak fractions at retention time of 32.9-35.1 min were collected, and concentrated using a rotary evaporator under vacuum in nitrogen. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried samples were frozen in a refrigerator, to give 1-palmitin-2,3-diolein.

(71) HR-EI-MS: m/z=858.7672 (Calcd.=858.7676, C.sub.55H.sub.102O.sub.6), Degree of unsaturation=5.

(72) IR (KBr film): 1747, 1166, 1095; 2926, 2854, 722; 3003, 1654 cm.sup.1 (weak).

(73) .sup.1H-NMR data are shown in Table 8.

(74) .sup.13C-NMR data are shown in Table 9.

Example 19 Preparation of 1,3-dipalmitin-2-olein

(75) Coix seed oil solution prepared in tetrahydrofuran (750 mg/mL) was preliminarily separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP C18 (2), 50*250 mm, 5 m, 100 ; Mobile phase: acetonitrile/tetrahydrofuran=78:22 (v/v); Injection volume 2 ml; Flow rate: 80 mL/min; UV detection wavelength: 205 nm/280 nm). Peak fractions at retention time of 29-38 min were collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator to give a crude product.

(76) The second purification was proceeded on Venusil XBP C18(L) Column (30*150 mm, 5 m 150 ) with acetonitrile:dichloromethane (65/35) as the mobile phase in a flow rate of 32 mL/min. A solution of the above crude product (10 mg/mL) was prepared in dichloromethane, and the injection volume for each separation was 2 mL. UV detection was conducted at 205 nm/280 nm. Peak fraction at retention time of 15 min was collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried sample, 1,3-dipalmitin-2-olein, was frozen in a refrigerator.

(77) HR-EI-MS: m/z=832.7542 (Calcd.=832.7566, C.sub.53H.sub.100O.sub.6), Degree of unsaturation=4.

(78) IR (KBr film): 1747, 1166, 1095; 2925, 2854, 722; 3003, 1654 cm.sup.1 (week).

(79) .sup.1H-NMR (CDCl.sub.3) data are shown in Table 10.

(80) TABLE-US-00010 TABLE 10 .sup.1H-NMR spectral data of the compound of Example 19 Amount of H, Spike & Position Chemical shift coupling constant HCC 5.34 2H, m CHOCO 5.26 1H, m CH.sub.2OCO 4.29 2H, dd, 12.0, 4.4 CH.sub.2OCO 4.14 2H, d, 12.0, 6.0 CH2COO 2.31 2H, t, 7.6 CH2COO 2.31 4H, t, 7.6 CH.sub.2CH 2.00 4H, m CH.sub.2CH.sub.2COO 1.60 6H, m CH.sub.2 1.27 68H, m CH.sub.3 0.87 9H, t, 6.8

(81) .sup.13C-NMR (CDCl.sub.3) data are shown in Table 11.

(82) TABLE-US-00011 TABLE 11 .sup.13C-NMR (CDCl.sub.3) spectral data of the compound of Example 19 Position Chemical shift C1 1 173.282 2 172.839 3 173.282 C2 1 34.026 2 34.179 3 34.026 C3 1 24.848 2 24.848 3 24.848 C4 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C5 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C6 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C7 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C8 1 29.750~29.040 2 27.159 3 29.750~29.040 C9 1 29.750~29.040 2 129.687 3 29.750~29.040 C10 1 29.750~29.040 2 129.999 3 29.750~29.040 C11 1 29.750~29.040 2 27.205 3 29.750~29.040 C12 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C13 1 29.750~29.040 2 29.750~29.040 3 29.750~29.040 C14 1 31.895 2 29.750~29.040 3 31.895 C15 1 22.672 2 29.750~29.040 3 22.672 C16 1 14.099 2 31.895 3 14.099 C17 2 22.672 C18 2 14.099 CHO 68.852 CH2O 62.076

Example 20 Preparation of 1,2-Diolein-3-Stearin

(83) Coix seed oil solution prepared in tetrahydrofuran (750 mg/mL) was preliminarily separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP C18 (2), 50*250 mm, 5 m, 100 ; Mobile phase: acetonitrile/tetrahydrofuran=78:22 (v/v); Injection volume 2 ml (1.5 g); Flow rate: 80 mL/min; UV detection wavelength: 205 nm/280 nm). Peak fractions at retention time of 29-38 min were collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator, to give a crude product.

(84) The second purification was proceeded on Venusil XBP C18(L) Column (30*150 mm, 5 m 150 ) with acetonitrile:dichloromethane (65/35) as the mobile phase in a flow rate of 32 mL/min. A solution of the above crude product (10 mg/mL) was prepared in dichloromethane, and the injection volume for each separation was 2 mL. UV detection was conducted at 205 nm/280 nm. Peak fraction at retention time of 17 min was collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried sample, 1,2-diolein-3-stearin, was frozen in a refrigerator.

(85) HR-EI-MS: m/z=886.8011 (Calcd.=886.7991, C.sub.57H.sub.106O.sub.6), Degree of unsaturation=5.

(86) IR (KBr film): 1747, 1166, 1095; 2926, 2856, 722; 3003, 1654 cm.sup.1 (week).

(87) .sup.1H-NMR (CDCl.sub.3) data are shown in Table 12.

(88) TABLE-US-00012 TABLE 12 .sup.1H-NMR spectral data of the compound of Example 20 Amount of H, Spike & Position Chemical shift coupling constant HCC 5.35 4H, m -CHOCO 5.26 1H, m -CH.sub.2OCO 4.29 2H, dd, 11.6, 4.0 -CH.sub.2OCO 4.14 2H, d, 12.0, 6.0 CH2COO 2.31 2H, t, 7.6 CH2COO 2.31 4H, t, 7.6 CH.sub.2CH 2.01 8H, m CH.sub.2CH.sub.2COO 1.60 6H, m CH.sub.2 1.27 74H, m CH.sub.3 0.87 9H, t, 6.8

(89) .sup.13C-NMR (CDCl.sub.3) data are shown in Table 13.

(90) TABLE-US-00013 TABLE 13 .sup.13C-NMR (CDCl.sub.3) spectral data of the compound of Example 20 Position Chemical shift C1 1 173.258 2 172.839 3 173.282 C2 1 34.028 2 34.179 3 34.028 C3 1 24.848 2 24.848 3 24.848 C4 1 30.00~29.00 2 30.00~29.00 3 30.00~29.00 C5 1 30.00~29.00 2 30.00~29.00 3 30.00~29.00 C6 1 30.00~29.00 2 30.00~29.00 3 30.00~29.00 C7 1 30.00~29.00 2 30.00~29.00 3 30.00~29.00 C8 1 27.159 2 27.159 3 29.00~30.00 C9 1 129.687 2 129.669 3 29.00~30.00 C10 1 129.999 2 129.999 3 29.00~30.00 C11 1 27.206 2 27.206 3 29.00~30.00 C12 1 29.00~30.00 2 29.00~30.00 3 29.00~30.00 C13 1 29.00~30.00 2 29.00~30.00 3 29.00~30.00 C14 1 29.00~30.00 2 29.00~30.00 3 31.895 C15 1 29.00~30.00 2 29.00~30.00 3 22.672 C16 1 31.895 2 31.895 3 31.895 C17 1 22.672 2 22.672 3 22.672 C18 1 14.099 2 14.099 3 14.099 -CHO 68.852 -CH2O 62.076

Example 21 Preparation of 1-Olein-2-Linolein-3-Stearin

(91) Coix seed oil solution prepared in tetrahydrofuran (750 mg/mL) was preliminarily separated in CHEETAH-HP100 preparative high performance liquid chromatography (Column: Venusil XBP C18 (2), 50*250 mm, 5 m, 100 ; Mobile phase: acetonitrile/tetrahydrofuran=78:22 (v/v); Flow rate of 80 mL/min; Injection volume 2 ml (1.5 g); UV detection wavelength: 205 nm/280 nm). Peak fractions at retention time of 29-38 min were collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator, to give a crude product.

(92) The second purification was proceeded on Venusil XBP C18(L) Column (30*150 mm, 5 m 150 ) with acetonitrile:dichloromethane (65/35) as the mobile phase in a flow rate of 32 mL/min. A solution of the above crude product (10 mg/mL) was prepared in dichloromethane, and the injection volume for each separation was 2 mL. UV detection was conducted at 205 nm/280 nm. Peak fraction at retention time of 19 min was collected, and concentrated under vacuum in nitrogen atmosphere in a rotary evaporator. Residues were transferred with chloroform to a 10 mL vial, and dried in a vacuum oven at 35 C. for 6 h. After filling with nitrogen, the dried sample, 1-olein-2-linolein-3-stearin, was frozen in a refrigerator.

(93) HR-EI-MS: m/z=884.7832 (Calcd.=884.7848, C.sub.57H.sub.104O.sub.6), Degree of unsaturation=6.

(94) IR (KBr film): 1747, 1164, 1098; 2925, 2855, 723; 3008, 1655 cm.sup.1 (week).

(95) .sup.1H-NMR (CDCl.sub.3) data are shown in Table 14.

(96) TABLE-US-00014 TABLE 14 .sup.1H-NMR spectral data of the compound of Example 21 Amount of H, Spike & Position Chemical shift coupling constant HCC 5.35 6H, m CHOCO 5.26 1H, m CH.sub.2OCO 4.29 2H, dd, 11.4, 4.2 CH.sub.2OCO 4.14 2H, d, 12.0, 6.0 CH.sub.2COO 2.32 2H, m CH.sub.2COO 2.30 4H, m CH.sub.2CH 2.03 8H, m CHCHCH.sub.2CHCH 2.77 2H, t, 6.6 CH.sub.2CH.sub.2COO 1.61 6H, m CH.sub.2 1.38~1.26 64H, m CH.sub.3 0.88 9H, t, 6.6

(97) .sup.13C-NMR (CDCl.sub.3) data are shown in Table 15.

(98) TABLE-US-00015 TABLE 15 .sup.13C-NMR (CDCl.sub.3) spectral data of the compound of Example 21 Position Chemical shift C1 1 173.25 2 172.83 3 173.25 C2 1 34.03 2 34.18 3 34.04 C3 1 24.86 .sup.2.sup.b 24.86 3 24.86 C4 1 29.04~29.76 .sup.2.sup.b 29.04~29.76 3 29.04~29.76 C5 1 29.04~29.76 .sup.2.sup.b 29.04~29.76 3 29.04~29.76 C6 1 29.04~29.76 .sup.2.sup.b 29.04~29.76 3 29.04~29.76 C7 1 29.04~29.76 .sup.2.sup.b 29.04~29.76 3 29.04~29.76 C8 1 29.04~29.76 2 27.19 3 29.04~29.76 C9 1 129.70 2 130.01 3 29.04~29.76 C10 1 129.97 2 128.08 3 29.04~29.76 C11 1 27.19 2 25.62 3 29.04~29.76 C12 1 29.04~29.76 2 127.88 3 29.04~29.76 C13 1 29.04~29.76 2 130.22 3 29.04~29.76 C14 1 31.89 2 29.04~29.76 3 31.89 C15 1 22.67 .sup.2.sup.b 22.56 3 22.67 C16 1 31.91 2 31.52 3 31.89 C17 1 22.67 2 22.56 3 22.67 C18 1 14.09 2 14.05 3 14.09 CHO 68.89 2CH.sub.2O 62.08

Example 22 Preparation of Coix Seed Oil Injection of the Invention

(99) Formulation:

(100) TABLE-US-00016 Coix seed oil 100 g Soybean lecithin for injection 10 g Glycerin for injection 15 g Water for injection added to 1000 mL

(101) wherein, the Coix seed oil contains following ingredients:

(102) TABLE-US-00017 1,3-diolein 0.50% 1-linolein-3-olein 1.31% 1,2-diolein 0.30% 1-olein-2-linolein 0.95% 1,2-dilinolein 0.41% Trilinolein 6.10% 1-Olein-2,3-dilinolein 16.18% 1-Palmitin-2,3-dilinolein 6.56% 1,3-Diolein-2-linolein 16.69% 1-Palmitin-2-linolein-3-olein 12.96% 1,3-Dipalmitin-2-linolein 2.88% Triolein 18.30% 1-Palmitin-2,3-diolein 10.18% 1-olein-2-linolein-3-stearin 1.72% 1,3-dipalmitin-2-olein 1.88% 1,2-diolein-3-stearin 1.60%

(103) Process:

(104) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(105) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 60 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 6 MPa and the high pressure was 30 MPa. The homogenization was repeated for 4 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 8.5.

(106) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 23 Preparation of Coix Seed Oil Injection of the Invention

(107) Formulation:

(108) TABLE-US-00018 Coix seed oil 300 g Soybean lecithin acceptable for injection 40 g Glycerin acceptable for injection 50 g Water for injection added to 1000 mL

(109) wherein, the Coix seed oil contains following ingredients:

(110) TABLE-US-00019 1,3-diolein 0.40% 1-linolein-3-olein 1.05% 1,2-diolein 0.24% 1-olein-2-linolein 0.76% 1,2-dilinolein 0.33% Trilinolein 4.87% 1-Olein-2,3-dilinolein 17.88% 1-Palmitin-2,3-dilinolein 5.25% 1,3-Diolein-2-linolein 15.13% 1-Palmitin-2-linolein-3-olein 10.26% 1,3-Dipalmitin-2-linolein 3.05% Triolein 20.46% 1-Palmitin-2,3-diolein 11.50% 1-olein-2-linolein-3-stearin 1.95% 1,3-dipalmitin-2-olein 2.16% 1,2-diolein-3-stearin 1.84%

(111) Process:

(112) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(113) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 70 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 12 MPa and the high pressure was 45 MPa. The homogenization was repeated for 3 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 7.1.

(114) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 24 Preparation of Coix Seed Oil Injection of the Invention

(115) Formulation:

(116) TABLE-US-00020 Coix seed oil 200 g Soybean lecithin for injection 25 g Glycerin acceptable for injection 30 g Water for injection added to 1000 mL

(117) wherein, the Coix seed oil contains following ingredients:

(118) TABLE-US-00021 1,3-diolein 0.45% 1-linolein-3-olein 1.18% 1,2-diolein 0.27% 1-olein-2-linolein 0.86% 1,2-dilinolein 0.37% Trilinolein 5.47% 1-Olein-2,3-dilinolein 18.69% 1-Palmitin-2,3-dilinolein 6.01% 1,3-Diolein-2-linolein 18.19% 1-Palmitin-2-linolein-3-olein 14.11% 1,3-Dipalmitin-2-linolein 2.60% Triolein 16.25% 1-Palmitin-2,3-diolein 9.11% 1-olein-2-linolein-3-stearin 1.88% 1,3-dipalmitin-2-olein 2.09% 1,2-diolein-3-stearin 1.76%

(119) Process:

(120) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(121) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 65 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 10 MPa and the high pressure was 30 MPa. The homogenization was repeated for 5 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 4.8.

(122) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 25 Preparation of Coix Seed Oil Injection of the Invention

(123) Formulation:

(124) TABLE-US-00022 Coix seed oil 150 g Soybean lecithin acceptable for injection 35 g Glycerin acceptable for injection 30 g Water for injection added to 1000 mL

(125) wherein, the Coix seed oil contains following ingredients:

(126) TABLE-US-00023 1,3-diolein 0.49% 1-linolein-3-olein 1.28% 1,2-diolein 0.29% 1-olein-2-linolein 0.93% 1,2-dilinolein 0.40% Trilinolein 5.96% 1-Olein-2,3-dilinolein 16.58% 1-Palmitin-2,3-dilinolein 6.43% 1,3-Diolein-2-linolein 16.20% 1-Palmitin-2-linolein-3-olein 12.57% 1,3-Dipalmitin-2-linolein 2.79% Triolein 17.69% 1-Palmitin-2,3-diolein 9.87% 1-olein-2-linolein-3-stearin 1.75% 1,3-dipalmitin-2-olein 1.92% 1,2-diolein-3-stearin 1.66%

(127) Process:

(128) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(129) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 68 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 7 MPa and the high pressure was 35 MPa. The homogenization was repeated for 3 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 6.8.

(130) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 26 Preparation of Coix Seed Oil Capsule of the Invention

(131) Formulation:

(132) TABLE-US-00024 Coix seed oil 200 g Vitamine E 0.20 g to give 1000 capsules

(133) wherein, the Coix seed oil contains following ingredients:

(134) TABLE-US-00025 1,3-diolein 0.51% 1-linolein-3-olein 1.34% 1,2-diolein 0.31% 1-olein-2-linolein 0.97% 1,2-dilinolein 0.42% Trilinolein 6.20% 1-Olein-2,3-dilinolein 15.93% 1-Palmitin-2,3-dilinolein 6.69% 1,3-Diolein-2-linolein 16.87% 1-Palmitin-2-linolein-3-olein 13.09% 1,3-Dipalmitin-2-linolein 2.91% Triolein 18.42% 1-Palmitin-2,3-diolein 10.27% 1-olein-2-linolein-3-stearin 1.67% 1,3-dipalmitin-2-olein 1.86% 1,2-diolein-3-stearin 1.56%

(135) Process:

(136) Glue formulation: Gelatin, purified water, glycerin and 10% ethylparaben solution were weighed at a weight ratio of 1:1.2:0.8:0.01. Glycerin, purified water and 10% ethylparaben solution were sequentially added into a glue melting tank and heated to 70 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 60 C. for use.

(137) Drug liquid formulation: Formulated amount of Coix seed oil and vitamin E were added into an ingredient tank and stirred constantly until thoroughly mixed.

(138) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 18 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 27 Preparation of Coix Seed Oil Capsule of the Invention

(139) Formulation:

(140) TABLE-US-00026 Coix seed oil 800 g Tween 80 0.60 g to give 1000 capsules

(141) wherein, the Coix seed oil contains following ingredients:

(142) TABLE-US-00027 1,3-diolein 0.55% 1-linolein-3-olein 1.44% 1,2-diolein 0.33% 1-olein-2-linolein 1.05% 1,2-dilinolein 0.45% Trilinolein 6.69% 1-Olein-2,3-dilinolein 14.75% 1-Palmitin-2,3-dilinolein 7.21% 1,3-Diolein-2-linolein 14.92% 1-Palmitin-2-linolein-3-olein 11.55% 1,3-Dipalmitin-2-linolein 3.14% Triolein 19.86% 1-Palmitin-2,3-diolein 11.08% 1-olein-2-linolein-3-stearin 1.50% 1,3-dipalmitin-2-olein 1.70% 1,2-diolein-3-stearin 1.43%

(143) Process:

(144) Glue formulation: Gelatin, purified water, glycerin and benzoic acid were weighed at a weight ratio of 1:1.2:0.8:0.01. Glycerin, purified water and benzoic acid were sequentially added into a glue melting tank and heated to 90 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 56 C. for use.

(145) Drug liquid formulation: Formulated amount of Coix seed oil and Tween 80 were added into an ingredient tank and stirred constantly until thoroughly mixed.

(146) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 26 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 28 Preparation of Coix Seed Oil Capsule of the Invention

(147) Formulation:

(148) TABLE-US-00028 Coix seed oil 500 g Vitamine E 0.40 g to give 1000 capsules

(149) wherein, the Coix seed oil contains following ingredients:

(150) TABLE-US-00029 1,3-diolein 0.58% 1-linolein-3-olein 1.14% 1,2-diolein 0.35% 1-olein-2-linolein 0.83% 1,2-dilinolein 0.47% Trilinolein 6.99% 1-Olein-2,3-dilinolein 13.00% 1-Palmitin-2,3-dilinolein 7.54% 1,3-Diolein-2-linolein 19.02% 1-Palmitin-2-linolein-3-olein 14.75% 1,3-Dipalmitin-2-linolein 3.28% Triolein 15.96% 1-Palmitin-2,3-diolein 9.70% 1-olein-2-linolein-3-stearin 1.38% 1,3-dipalmitin-2-olein 1.52% 1,2-diolein-3-stearin 1.29%

(151) Process:

(152) Glue formulation: Gelatin, purified water, glycerin and potassium sorbate were weighed at a weight ratio of 1:0.9:0.6:0.005. Glycerin, purified water and potassium sorbate were sequentially added into a glue melting tank and heated to 80 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 62 C. for use.

(153) Drug liquid formulation: Formulated amount of Coix seed oil and Vitamin E were added into an ingredient tank and stirred constantly until thoroughly mixed.

(154) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 28 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 29 Preparation of Coix Seed Oil Capsule of the Invention

(155) Formulation:

(156) TABLE-US-00030 Coix seed oil 600 g Tween 80 0.3 g to give 1000 capsules

(157) wherein, the Coix seed oil contains following ingredients:

(158) TABLE-US-00031 1,3-diolein 0.45% 1-linolein-3-olein 1.26% 1,2-diolein 0.27% 1-olein-2-linolein 0.88% 1,2-dilinolein 0.36% Trilinolein 6.15% 1-Olein-2,3-dilinolein 18.01% 1-Palmitin-2,3-dilinolein 6.66% 1,3-Diolein-2-linolein 16.77% 1-Palmitin-2-linolein-3-olein 12.89% 1,3-Dipalmitin-2-linolein 2.88% Triolein 18.30% 1-Palmitin-2,3-diolein 8.69% 1-olein-2-linolein-3-stearin 1.81% 1,3-dipalmitin-2-olein 2.08% 1,2-diolein-3-stearin 1.81%

(159) Process:

(160) Glue formulation: Gelatin, purified water, glycerin and chlorhexidine acetate were weighed at a weight ratio of 1:1.0:0.5:0.008. Glycerin, purified water and chlorhexidine acetate were sequentially added into a glue melting tank and heated to 85 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 56 C. for use.

(161) Drug liquid formulation: Formulated amount of Coix seed oil and Tween 80 were added into an ingredient tank and stirred constantly until thoroughly mixed.

(162) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 30 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 30 Preparation of Coix Seed Oil Injection of the Invention

(163) Formulation:

(164) TABLE-US-00032 Coix seed oil 100 g Soybean lecithin for injection 10 g Glycerin for injection 15 g Water for injection added to 1000 mL

(165) wherein, the Coix seed oil contains following ingredients:

(166) TABLE-US-00033 1,3-diolein 0.42% 1-linolein-3-olein 1.25% 1,2-diolein 0.25% 1-olein-2-linolein 0.81% 1,2-dilinolein 0.35% Trilinolein 6.85% 1-Olein-2,3-dilinolein 18.24% 1-Palmitin-2,3-dilinolein 5.74% 1,3-Diolein-2-linolein 15.01% 1-Palmitin-2-linolein-3-olein 10.95% 1,3-Dipalmitin-2-linolein 2.88% Triolein 20.75% 1-Palmitin-2,3-diolein 10.18% 1-olein-2-linolein-3-stearin 1.92% 1,3-dipalmitin-2-olein 2.11% 1,2-diolein-3-stearin 1.84%

(167) Process:

(168) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin for injection was added. Then water for injection was added to a specified amount, and the mixture was stirred to give a water phase.

(169) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 60 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 7 MPa and the high pressure was 26 MPa. The homogenization was repeated for 5 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 6.8.

(170) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 31 Preparation of Coix Seed Oil Injection of the Invention

(171) Formulation:

(172) TABLE-US-00034 Coix seed oil 300 g Soybean lecithin acceptable for injection 40 g Glycerin acceptable for injection 50 g Water for injection added to 1000 mL

(173) wherein, the Coix seed oil contains following ingredients:

(174) TABLE-US-00035 1,3-diolein 0.46% 1-linolein-3-olein 1.20% 1,2-diolein 0.28% 1-olein-2-linolein 0.90% 1,2-dilinolein 0.38% Trilinolein 5.71% 1-Olein-2,3-dilinolein 15.11% 1-Palmitin-2,3-dilinolein 6.02% 1,3-Diolein-2-linolein 16.30% 1-Palmitin-2-linolein-3-olein 14.20% 1,3-Dipalmitin-2-linolein 3.20% Triolein 19.91% 1-Palmitin-2,3-diolein 9.22% 1-olein-2-linolein-3-stearin 1.78% 1,3-dipalmitin-2-olein 2.01% 1,2-diolein-3-stearin 1.70%

(175) Process:

(176) To a formulated amount of soybean lecithin acceptable for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin acceptable for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(177) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 70 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 11 MPa and the high pressure was 48 MPa. The homogenization was repeated for 6 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 7.5.

(178) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 32 Preparation of Coix Seed Oil Injection of the Invention

(179) Formulation:

(180) TABLE-US-00036 Coix seed oil 200 g Soybean lecithin for injection 25 g Glycerin acceptable for injection 30 g Water for injection added to 1000 mL

(181) wherein, the Coix seed oil contains following ingredients:

(182) TABLE-US-00037 1,3-diolein 0.50% 1-linolein-3-olein 1.31% 1,2-diolein 0.30% 1-olein-2-linolein 0.95% 1,2-dilinolein 0.41% Trilinolein 6.18% 1-Olein-2,3-dilinolein 17.26% 1-Palmitin-2,3-dilinolein 6.51% 1,3-Diolein-2-linolein 15.45% 1-Palmitin-2-linolein-3-olein 12.83% 1,3-Dipalmitin-2-linolein 2.81% Triolein 19.33% 1-Palmitin-2,3-diolein 9.95% 1-olein-2-linolein-3-stearin 1.71% 1,3-dipalmitin-2-olein 1.97% 1,2-diolein-3-stearin 1.69%

(183) Process:

(184) To a formulated amount of soybean lecithin for injection was added an appropriate amount of water for injection. The mixture was dispersed with a high shear dispersing emulsifier into a dispersion without bulks or granules. Formulated amount of glycerin acceptable for injection was added. Then water for injection is added to a specified amount, and the mixture was stirred to give a water phase.

(185) A formulated amount of Coix seed oil was weighed. The weighed oil and the water phase prepared above were heated separately to 65 C., then mixed and emulsified in a high pressure homogenizer, in which the low pressure was 8 MPa and the high pressure was 40 MPa. The homogenization was repeated for 4 times until the amount of particles below 2 m was no less than 95% and particles above 5 m were undetectable. If necessary, NaOH or HCl was used to adjust the pH to 6.5.

(186) The resulting homogeneous emulsion was filtered by nitrogen pressure through a microporous filter of 3 m or less, then filled under nitrogen, and finally sterilized and cooled to afford the injection.

Example 33 Preparation of Coix Seed Oil Capsule of the Invention

(187) Formulation:

(188) TABLE-US-00038 Coix seed oil 200 g Vitamine E 0.20 g to give 1000 capsules

(189) wherein, the Coix seed oil contains following ingredients:

(190) TABLE-US-00039 1,3-diolein 0.52% 1-linolein-3-olein 1.40% 1,2-diolein 0.32% 1-olein-2-linolein 1.01% 1,2-dilinolein 0.43% Trilinolein 6.51% 1-Olein-2,3-dilinolein 14.09% 1-Palmitin-2,3-dilinolein 6.84% 1,3-Diolein-2-linolein 17.65% 1-Palmitin-2-linolein-3-olein 13.56% 1,3-Dipalmitin-2-linolein 3.07% Triolein 18.10% 1-Palmitin-2,3-diolein 10.80% 1-olein-2-linolein-3-stearin 1.59% 1,3-dipalmitin-2-olein 1.73% 1,2-diolein-3-stearin 1.49%

(191) Process:

(192) Glue formulation: Gelatin, purified water, glycerin and 10% ethylparaben solution were weighed at a weight ratio of 1:1.2:0.8:0.01. Glycerin, purified water and 10% ethylparaben solution were sequentially added into a glue melting tank and heated to 70 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 59 C. for use.

(193) Drug liquid formulation: Formulated amount of Coix seed oil and Vitamin E were added into an ingredient tank and stirred constantly until thoroughly mixed.

(194) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 16 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 34 Preparation of Coix Seed Oil Capsule of the Invention

(195) Formulation:

(196) TABLE-US-00040 Coix seed oil 800 g Tween 80 0.60 g to give 1000 capsules

(197) wherein, the Coix seed oil contains following ingredients:

(198) TABLE-US-00041 1,3-diolein 0.56% 1-linolein-3-olein 1.11% 1,2-diolein 0.34% 1-olein-2-linolein 0.91% 1,2-dilinolein 0.46% Trilinolein 6.71% 1-Olein-2,3-dilinolein 16.31% 1-Palmitin-2,3-dilinolein 7.25% 1,3-Diolein-2-linolein 18.50% 1-Palmitin-2-linolein-3-olein 11.90% 1,3-Dipalmitin-2-linolein 2.63% Triolein 17.14% 1-Palmitin-2,3-diolein 11.21% 1-olein-2-linolein-3-stearin 1.42% 1,3-dipalmitin-2-olein 1.60% 1,2-diolein-3-stearin 1.31%

(199) Process:

(200) Glue formulation: Gelatin, purified water, glycerin and benzoic acid were weighed at a weight ratio of 1:1.2:0.8:0.01. Glycerin, purified water and benzoic acid were sequentially added into a glue melting tank and heated to 90 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 60 C. for use.

(201) Drug liquid formulation: Formulated amount of Coix seed oil and Tween 80 were added into an ingredient tank and stirred constantly until thoroughly mixed.

(202) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 26 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.

Example 35 Preparation of Coix Seed Oil Capsule of the Invention

(203) Formulation:

(204) TABLE-US-00042 Coix seed oil 500 g Vitamine E 0.40 g to give 1000 capsules

(205) wherein, the Coix seed oil contains following ingredients:

(206) TABLE-US-00043 1,3-diolein 0.57% 1-linolein-3-olein 1.21% 1,2-diolein 0.34% 1-olein-2-linolein 0.86% 1,2-dilinolein 0.46% Trilinolein 5.13% 1-Olein-2,3-dilinolein 16.03% 1-Palmitin-2,3-dilinolein 7.25% 1,3-Diolein-2-linolein 18.61% 1-Palmitin-2-linolein-3-olein 12.03% 1,3-Dipalmitin-2-linolein 3.01% Triolein 18.60% 1-Palmitin-2,3-diolein 11.21% 1-olein-2-linolein-3-stearin 1.39% 1,3-dipalmitin-2-olein 1.53% 1,2-diolein-3-stearin 1.30%

(207) Process:

(208) Glue formulation: Gelatin, purified water, glycerin and potassium sorbate were weighed at a weight ratio of 1:0.9:0.6:0.005. Glycerin, purified water and potassium sorbate were sequentially added into a glue melting tank and heated to 80 C. Then gelatin was added and constantly stirred under vacuum until the gelatin was completely dissolved. The glue was filtered and stored at 62 C. for use.

(209) Drug liquid formulation: Formulated amount of Coix seed oil and Vitamin E were added into an ingredient tank and stirred constantly until thoroughly mixed.

(210) Capsule pressing: Proper pellet dies were chosen according to the capsule size. Capsules were pressed under a temperature of 20 C. and a relative humidity of less than 35%, then shaped and dried. After excluding capsules of abnormal size, normal capsules were washed with 95% medicinal ethanol and dried continuously till the moisture content was less than 12%. Unqualified capsules were removed by visual inspection, and the final products were printed and packaged.