GB2366565A

GB2366565A - Zingiber officinale extract

Info

Publication number: GB2366565A
Application number: GB0114693A
Authority: GB
Inventors: Tian-Shung Wu; Sheng-Chu Kuo; Che-Ming Teng; Feng-Nien Ko
Original assignee: Pharmaceutical Industry Technology and Development Center
Current assignee: Pharmaceutical Industry Technology and Development Center
Priority date: 2000-07-12
Filing date: 2001-06-15
Publication date: 2002-03-13
Anticipated expiration: 2021-06-15
Also published as: DE10128266B4; CH692272A5; DE10128266A1; JP2002047195A; TWI290469B; GB2366565B; GB0114693D0; FR2811573B1; JP4052819B2; FR2811573A1

Abstract

A method of preparing an extract from <I>Zingiber officinale</I>, which is potent in anti-inflammation, anti-platelet aggregation and anti-fungal activity, includes the following steps: preparing a crude liquid from rhizomes of ginger by extraction with an organic solvent or supercritical CO<SB>2</SB> or by distillation with steam; introducing the crude liquid to a reverse phase chromatography column, and eluting the column with water, a first eluent and a second eluent having a polarity weaker than that of the first eluent but stronger than that of chloroform, so that a first eluate resulting from elution of the first eluent and a second eluate resulting from elution of the second eluent are obtained; removing the first eluent and the second eluent from the first eluate and the second eluate by evaporation, respectively, so that a first concentrated eluate and a second concentrated eluate are obtained as the potent extract. The final product or the crude liquid obtained in the first step may be incorporated into a pharmaceutical composition together with a suitable carrier or diluent.

Description

2366565 Method of Preparing an Extract Potent in Anti-inflammation,

Anti-platelet Aggregation and Anti-fungal Activity from Zingiber Officinale and Pharmaceutical Compositions Containing Said Extract

Field of the Invention

The present invention is related to a method of preparing an extract potent in anti-inflammation, anti-platelet aggregation and anti-fungal activity from Zingiber officinale.

10 Background of the Invention

Chinese crude drugs or spices eg. Zingiber officinale, Eugenia caryophyllata, Allium sativum, have been used in medicine and in flavoring foods. Crude ginger is used as an anti-emetic and expectorant, an antitussive and accelerator of the digestive organs. Semi-dried old crude 15 ginger is also used for stomachache, chest pain, low back pain, cough, common cold and as a cure for a form of edema being called "stagnate of water". Zingerone is the major component which accounts for the spicy character of ginger; gingerol and shogaol are other pungent components in ginger. Gingerol has cardio-tonic action, suppresses the contraction of 20 isolated portal veins in mice, and modulates the eicosanoid-induced contraction of mouse and rat blood vessels. Shogaol exhibits pressor response. Both gingerol and shogaol are mutagenic, whereas zinger and zingerone have been found to exhibit antimutagenic activity. Shogaolhas inhibitory activity on the carrageenin-induced paw edema and platelet aggregation [US patent No. 5,804,603, Background of the Invention].

Heretofore, many reports have shown that Zingiber officinale exhibits various physiological activities. Typical examples include a cancer metastasis suppressing agent disclosed in Japan patent publication No. 7 5 258104; a synthesis promoter for neurotropic factor, which is effective for nerve deteriorative diseases such as Alzheimer's dementia or Parkinson's disease, disclosed in Japan patent publication No. 7-25777; an anti rheumatic agent disclosed in Japan patent publication No. 6-293653, US patent Nos. 5,494,668 and 5,683,698; an anti-microbial composition 10 disclosed in Japan patent publication No. 6-227931; and an analgesic composition disclosed in Japan patent publication No. 6-107556. Ginger contains 1-4% essential oil (oleoresin). During the last 45 years many chemical investigations have been carried out on the constituents of the essential oil. Altogether more than 200 different volatiles have been 15 identified in essential oil wherein the pharmacological activity is confined. The essential oil contains a mixture of various terpenes as well as some other non-terpenoid compounds. Although this is mostly speculative, the experimental data and observations suggest that ginger inhibits both the cyclooxygenase and lypoxygenase products, i.e. it can be a dual inhibitor 20 of eicosanoid synthesis. In all 56 patients (28 with rheumatoid arthritis, 18 with osteoarthritis and 10 with muscular discomfort) used powdered ginger against their afflictions. Amongst the arthritis patients more than three-quarters experienced, to varying degrees, relief in pain and swelling. All the patients with muscular discomfort experienced relief in pain. None of the patients reported adverse effects during the period of ginger consumption which ranged from 3 months to 2.5 years. (Srivastava and Mustafa; Medical Hypotheses; 1992; 39 342-348) Non-steroidal anti-inflammatory drugs have three.major actions,. all of 5 which are related to inhibition of cyclo-oxygenase resulting in decreased formation of prostanoids. Firstly, an anti-inflammatory action achieved by reduced production of vasodilator prostaglandins (PGE2, PG12) which means less vasodilation and, indirectly less edema. Secondly, an analgesic effect achieved by reduced prostaglandin production (less 10 sensitization of nociceptic nerve endings to the inflammatory mediators bradykinin and 5-hydroxytryptamine). Thirdly, an antipyretic effect which is probably due to a decrease in the mediator PGE2generated in response to inflammatory pyrogens, much as interleukin-1. Since ginger inhibits prostanoid synthesis and also products of 5-lipoxygenase, its ameliorative 15 effects in arthritis and muscular discomforts could be related to reduced formation of prostanoids and leukotrienes. Because of such a possibility a decrease in the carageenan-induced edema formation in the rat's paw after 3 g of ginger extract administration has been demonstrated and the potency of the extract in the acute inflammation test appears to be 20 comparable to that exhibited by acetyl salicylic acid reported in the same study (Mascolo N. et al., Journal of Ethnopharmocology 1989, 27, 129140).

Dermatophytes, especially Trichophyton rubrum and Trichophyton mentagrophytes, are the usual pathogens of onychomycosis and tinea pedis [Roberts DT., British Journal of Dermatology. 141 Supple 56:1-4, 1999 Nov.; Roldan YB. et al., Mycoses, 43(5):181-3, 2000]. Pityrosporum ovale (Malassezia furfur) is the etiological agent of pityriasis vesicolor, Pityrosporum folliculitis and Malassezia intertrigo. Several studies 5 indicate a strong association of Pityrosporum ovale with seborrheic dermatitis and dandruff, a milder form of seborrheic dermatitis [Nenoff P. et al., Dermatology. 191(4):311-4, 1995; BulmerAC. et al., Mycopathologia, 147(2)63-5, 1999].

10 Summary of the Invention

The present invention provides extracts from rhizomes of ginger which show an activity in an in vitro anti-platelet aggregation test, an inhibitory activity on the carrageenin-induced paw edema, and antifungal activity against Trichophyton mentagrophytes and Pityrosporum ovale. The 15 extracts are prepared by extracting rhizomes of ginger with an organic solvent (such as ethyl ether, acetone, methanol and ethanol) or supercritical C02, or by steam distilling rhizomes of ginger to obtain a crude liquid, and subjecting said crude liquid to a reverse phase chromatography to obtain the extracts containing shogaols, gingerols and/or 20 dehydrogingerdione.

Detailed Description of the invention

As introduced in the Background of the Invention, ginger has been used for anti-inflammation and pain relief.

The present invention is to provide an effective method of preparing a product potent in anti-inflammation, in anti-platelet aggregation and in antifungal activity from rhizomes of ginger. The potent product prepared in accordance with the method of the present invention has a substantially 5 constant composition, so that the pharmacological effects thereof are definite.

The effective method of preparing product potent in anti-inflammation, in anti-platelet aggregation and in antifungal activity from rhizomes of ginger according to the present invention comprises the following steps:

10 a) preparing a crude liquid from rhizomes of ginger; b) introducing the crude liquid to a reverse phase chromatography column, and eluting the column with water, a first eluent and a second eluent in sequence, said second eluent having a polarity weaker than that of the first eluent but stronger than that of chloroform, so that a first eluate 15 resulting from elution of the first eluent and a second eluate resulting from elution of the second eluent are obtained; c) removing the first eluent from the first eluate by evaporation, so that a first concentrated eluate is obtained and is able to be used as the potent product; and 20 d) removing the second elue-nt from the second eluate by evaporation, so that a second concentrated eluate is obtained and is able to be used as the potent product; wherein step a) comprises steps i) to iv), or comprises step 1), step I'), or step I"), wherein said steps i) to iv) are:

i) shedding fresh rhizomes of ginger and filtering the resulting mixture to obtain a filtrate and a residue; ii) extracting the filtrate with a first organic solvent, recovering the resulting extraction solution of the first organic solvent, and evaporating the 5 first organic solvent from the extraction solution to obtain a first concentrated extraction solution; iii) extracting the residue with a second organic solvent, recovering the resulting extraction solution of the second organic solvent, and evaporating the second organic solvent from the extraction solution to 10 obtain a second concentrated extraction solution; and iv) combining the first concentrated extraction solution and the second concentrated extraction solution to obtain the crude liquid; said step 1) is:

1) extracting powder of dried rhizomes of ginger with the second 15 organic solvent, recovering the resulting extraction solution of the second organic solvent, and evaporating the second organic solvent from the extraction solution to obtain the crude liquid; said step I') is:

I') steam distilling powder of dried rhizomes of ginger, and 20 concentrating the resulting distillate by evaporation to obtain the crude liquid; and said step I") is:

I") extracting powder of dried rhizomes of ginger with supercritical C02, recovering the resulting extraction solution of the supercritical C02, and evaporatingC02 from the extraction solution to obtain the crude liquid.

The product potent in anti-inflammation, in anti-platelet aggregation and in antifungal activity prepared according to the method of the present invention preferably comprises 0 - 10 mg 6-shogaol per gram of the product, 5 1 - 150 mg 6-gingerol per gram of the product, and 0 - 40 mg 6 dehydrogingerdione per gram of the product.

The present invention also provides a pharmaceutical composition potent in anti-inflammation, in anti-platelet aggregation or in antifungal activity comprising a therapeutically effective amount of said crude liquid 10 prepared in step a) of the method of the present invention, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient.

The present invention also provides a pharmaceutical composition potent in anti-inflammation, in anti-platelet aggregation or in antifungal 15 activity comprising a therapeutically effective amount of said product prepared according to the method of the present invention, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient. Preferably, said product prepared according to the method of the present invention is the first concentrated 20 eluate prepared in step c). Alternatively, said product prepared according to the method of the present invention is the second concentrated eluate prepared in step d).

Preferably, said first eluent is methanol, and said second eluent is acetone.

Preferably, step a) of the method of the present invention comprises steps i) to iv).

Preferably, said first organic solvent is ethyl ether.

Preferably, said second organic solvent is acetone, methanol, ethanol 5 or a combination thereof. More preferably, said second organic solvent is acetone.

Preferably, step a) of the method of the present invention comprises step 1).

Preferably, step a) of the method of the present invention comprises 10 step I').

Preferably, step a) of the method of the present invention comprises step I").

A suitable reverse phase chromatography column for use in the method of the present invention includes (but not limited thereto) a reverse 15 phase chromatography column packed with a porous resin, for examples Diaion HP-20 (Mitsubishi Co.), Sephadex LH-20 (Pharmicia Co.) and RP18 (Nacalai tesque Co.).

The pharmaceutical composition potent in antifungal activity of the present invention is preferably applied topically, for examples as a 20 shampoo, a bath gel, soap, a body lotion, a body cream and a detergent.

Preferably, the pharmaceutical composition potent in antifungal activity of the present invention is used in the treatment of diseases associated with Trichophyton mentagrophytes or Pityrosporum ovale, including but are not limited to tinea pedis, tinea capitis, tinea cruris, tinea glabrosa, onychomycosis, pityriasis capitis, pityriasis vesicolor, pityrosporum folliculitis, seborrheic dermatitis and dandruff. In particular, the antifungal pharmaceutical composition of the present invention is in the form of a shampoo for use in the treatment of dandruff.

5 Without further elaboration, it is believed that the above description has adequately enabled the present invention. The following specific examples are, therefore, to be construed as merely illustrative, and not limitations on the remainder of the disclosure in any way whatsoever.

Determination of active ingredients In the following examples, high performance liquid chromatography (abbreviated as HPILC) was used to determine the active ingredients of the products prepared therein. HIPLC spectra were recorded on a HPILC instrument (HPILC Shimadzu I-C-1 OAT, Japan) using a Cosmosil 5C-1 8 15 column (250 mm x 4.6 mm, packed with particles having 5 tm diameter) by an elution method. An HIPILC sample was prepared by diluting an appropriate amount of a product with a mobile phase solution (hydrogen cyanide: water = 65: 35, V/V) to 25 ml, and filtered with a 0.25 Irn membrane. The filtrate was introduced into the HPLC column, and eluted 20 with the mobile phase solution. An UV detector (Shimadzu SPD-6AV, Japan) was used to detect the absorption of the eluate at 230 nm.

Example 1:

2100 g of fresh rhizomes of ginger were shredded and filtered to obtain a filtrate and a residue. 500 ml of the filtrate was extracted with 500 ml ethyl ether three times, the organic phase layers were separated from the aqueous phase layers, and combined. Ethyl ether was evaporated from the combined extraction solution in vacuo to obtain a 5 concentrated ethyl ether extraction product (I-OE). The ginger residue was extract with 3000 ml acetone three times, the extraction solutions were recovered by filtration, and combined. Acetone was evaporated from the combined extraction solution in vacuo to obtain a concentrated acetone extraction product (1-0) (14.5 g). To a reverse phase chromatography 10 column 300 mm x 30 mm packed with 180 g Diaion HP-20 resin having a diameter of 500 pm-800 pm 7 g of a mixture of the concentrate ethyl ether extraction product (I-OE) and the concentrated acetone extraction product (1-0) was injected. 1500 ml water, 2500 ml methanol, 2000 ml acetone and 2000 ml chloroform were used to carry out elution. The water eluate, 15 methanol eluate, acetone eluate and chloroform eluate were collected separately, and concentrated in vacuo to obtain 0.27 g concentrated water eluate (I-OW), 1.45 g concentrated methanol eluate (I-OM), 2.68 g concentrated acetone eluate (I-OA), and 0.83 g concentrated chloroform eluate (I-OC). The amounts (mg) of 6-shogaol, 6-gingerol and 6- 20 dehydrogingerdione per gram of the 1-0, I-OM and I-OA determined by HPLC are listed in Table 1.

Table 1

Content 1-0 I-OM I-OA (mg/g) 6-shogaol 1.10 0.14 1.15 0.0 6-gingerol 59.98 0.99 103.37 8.57 2.51 0.89 6-dehydrogingerdione 7.68 0.42 8.94 0.41 Example 2:

500 g of shade dried rhizomes of ginger were pulverized and the 5 resulting powder was extracted with 30 L acetone trice (each time with 10 Q. The three extraction solutions were combined together after filtration, and then concentrated in vacuo to obtain 24 g of concentrated acetone extraction product (11-0). To a reverse phase chromatography column packed with 600 g Diaion HP-20 resin 20 g of the concentrated acetone 10 extraction product (11-0) was injected, which was then eluted with 4 L water, 6.5 L methanol, 15 L acetone and 5 L chloroform in sequence. The water eluate, methanol eluate, acetone eluate and chloroform eluate were collected separately, and concentrated in vacuo to obtain 2.5 g concentrated water eluate (11-OW), 7.1 g concentrated methanol eluate (1115 OM), 6.9 g concentrated acetone eluate (H-OA), and 3.5 g concentrated chloroform eluate (11-OC). The amounts (mg) of 6-shogaol, 6-gingerol and 6-dehydrogingerdione per gram of the 11-0, 11-OM and 11-OA determined by HPLC are listed in Table 2.

Table 2

Content 11-0 11-OM 11-OA (mg/g) 6-shogaol 1.98 0.00 4.96 0.00 6-gingerol 43.06 0.84 70.87 1.85 2.54 0.00 6-dehydrogingerdione 9.33 0.85 19.15 4.57 2.35 0.28 Example 3:

Kg of shade dried rhizomes of ginger were pulverized and the 5 resulting powder was steam distilled for five hours. The distillate was concentrated in vacuo to obtain 410 g of concentrated distillate (111-0). To a reverse phase chromatography column packed with 600 g Diaion HP-20 resin 20 g of the concentrated distillate (111-0) was injected, which was then eluted with 4.5 L water, 4.5 L methanol, 3 L acetone and 5 L chloroform in 10 sequence. The water eluate, methanol eluate, acetone eluate and chloroform eluate were collected separately, and concentrated in vacuo to obtain 0.03 g concentrated water eluate (III-OW), 14.5 g concentrated methanol eluate (III-OM), 0.85 g concentrated acetone eluate (III-OA), and 0.2 g concentrated chloroform eluate (III-OC). The concentrated distillate 15 (111-0) contains no 6-shogaol, 6-gingerol and 6-dehydrogingerdione determined by HPLC.

Example 4:

g of powder of shade dried rhizomes of ginger was extracted with 1000 ml acetone at 500C for two hours. The extraction solution was separated and concentrated in vacuo (400C, 75 mmHg) to obtain a concentrated acetone extraction product (IV-0). The color and viscosity of the product (IV-0) together with its yield are listed in Table 3.

Example 5:

g of powder of shade dried rhizomes of ginger was steam distilled, and the oily distillate after being separated from the aqueous distillate was freeze dried to obtain an oily extract (V-0). The color and viscosity of the 10 oily extract (V-0) together with its yield are listed in Table 3 Example 6:

To 10 g of powder of shade dried rhizomes of ginger in a 250 ml extraction chamberCo2was introduced at a flow rate of 45 L/mIn, wherein 15 the chamber pressure was controlled at 2500 to 4000 psia with a high pressure pump (Model No. EK-1, LEWA Co., US) and the chamber temperature was maintained at 35-60'C with a heat exchanger (Model No.

H-2410, HOTEC Co., US) and an exterior circulation system. The extraction was stopped when the volume of C02 introduced reached 300 L, 20 and a supercritical C02 extraction product (VI-0) was obtained after evaporationof C02. The color and viscosity of the product (VI-0) together with its yield are listed in Table 3. The contents of pungent components determined by HPLC are listed in Table 4.

Table 3

IV-0 V-0 VI-0 L 87.6 80.4 96.3 A -9.1 -0.1 -9.6 B 31.1 9.6 22.0 Viscosity (cPs) 15.6 11.8 12.1 Yield (%) 3.8 2.2 3.9 the values of L, A, and B were determined by using a 190 color measuring system, (Nippon Denshoku Inc, Co., Ltd., Japan), wherein L represents lightness, A is the red/green difference and B is the yellow/blue difference.

Table 4

Content (mg/g) VI-0 6-shogaol 17.30 0.00 6-gingerol 26.29 0.00 6-dehydrogingerdione 19.20 1.19 Example 7: Antiplatelet assay 10 Blood, collected from the marginal ear vein of rabbits was mixed with EDTA (100 mM) in a volume ratio of 14:1 and centrifuged at 90 g for 10 min at room temperature to obtain platelet-rich plasma. The latter was further centrifuged at 500 g for 10 min, the upper plasma-rich layer was removed therefrom, and the remaining bottom layer was suspended with Tyrode's 15solution containing 2 mM EDTA but no calcium. The suspension was further centrifuged at 500 g for 10 min and the platelets were suspended with Tyrode's solution without EDTA. After centrifugation at the same conditions, the platelets were suspended with Tyrode's solution having the 5 following compositions (mM): NaCl (136.8), KCI (2.8), NaNCO, (11.9), MgCl2 (1.1), Nal-12POI (0.33), CaCl2 0 0), glucose (11.2) and borine serum albumin (0.35%). Platelet numbers were determined with a Coulter Counter (Model ZM) and adjusted to 4.5 x 10' platelets/ml.

Table 5. The inhibitory effects of ginger extracts on platelet aggregation induced by arachidonic acid and collagen') Concentration for 50% inhibitory effect (pg/ml) Ginger extra Ctb) Arachidonic acid Collagen 1-0 3.8 0.8 5.5 0.4 I-OM 1.7 0.3 2.7 0.4 11-0 3.1 0.5 6.5 1.2 11-OA 10.9 3.2 21.8 2.2 11-0c 6.9 0.7 16.6 4.3 11-OM 2-0 0.2 6.9 2.4 a) Platelets were incubated with ginger extracts or 0.5% DMSO (Control) at 370C for 3 min, then arachidonic acid (100 pM) or collagen (10 pg/ml) was added to trigger aggregation. Aspirin and Indomethacin are positive 15 controls. The percentage of inhibitory effect is calculated as follows:

ffidegree of inhibition of Control) - (degree of inhibition of ginger extract)] (degree of inhibition of Control)} x 100% Values are presented as mean S. E., n=3-6.

) Prepared in Examples 1 and 2 Example 8: Evaluation of inhibitory activity on the carrageenin-induced paw edema Inhibitory activity on the carrageenin-induced paw edema was conducted according to the method reported by Winter, C. A. et al. (Winter 5 C. A. et al., Proc. Soc. Exper. Biol. Med. 111: 544-547, 1962.). Male Wistar mice weighing 150 20 g without feeding for one night were injected at left rear paws thereof with 0.1 ml of 1% carrageenin suspension followed by rubbing test samples or vehicle as control on the left rear paws evenly (10 mg/paw). Three hours later, the volumes of the rear paws were 10 determined by using a volume scanner (Cat. #7150, UGO Basil, Italy), and the difference between the left rear paw and the right rear paw was used an index of the carrageenin-induced paw edema.

Table 6. Inhibitory activity on the carrageenin-induced paw edema of ginger extracts'9) Treatmentb) Dosage Inhibitory activity on the (mg/paw) carrageenin-induced paw edema M 1-0 10 18 I-OE 10 19 I-OM 10 29 I-OA 10 25 I.i. -0. ".- i.-, 0 1.. 8 II-OW 10 0 H-OM 10 26 11-OA 10 25 11-0c 10 8 -.. 1 11.1...... ..

111-0 III-OM 10 11 III-OA 10 15 [6] A e h yd ro g i n g e d i o n e 5 26 Inhibitory activity on the carrageenin-induced paw edema (%) was calculated as follows:

5 [(average degree of edema of mice in the control group)-(average degree of edema of mice in the test group)/(average degree of edema of mice in the control group)] x 100% Values are presented as mean S. E., n=3-6.

b) Prepared in Examples 1, 2 and 3 Example 9: Evaluation of inhibitory activity on Trichophyton mentagrophytes or Pityrosporum ovale AntiTrichophyton mentagrophytes assay Minimum inhibitory concentration (MIC) of ginger extract was determined and conducted according to the method previously described by Edwards, J. R. et a/. (Edwards, J. R. et al., Antimicrobial Agents Chemotherapy 33: 215-222, 1989). The test substance was dissolved and serially diluted in solvent (100% DMSO) to desired stock concentrations. For each concentration tested, a 0.01 mi aliquot was 5 added to a 48-well plate containing 0.99 ml of Potato Dextrose Broth (DIFCO, U.S.A.) with 101_104 CFU/ml of Trichophyton mentagrophytes (ATCC 9533). The plates were incubated at 28'C for 72 hours and then visually examined and scored. Vehicle-control was used as blank control. Each concentration was evaluated in duplicate. The results are shown in 10 Table 7.

Anti-Pityrosporum ovale assay Minimum inhibitory concentration (MIC) of ginger extract was determined and conducted by the method as mentioned above (Edwards, J. R. et al., Antimicrobial Agents Chemotherapy 33: 215-222, 1989). The 15 test substance was dissolved and serially diluted in solvent (100% DMSO) to desired stock concentrations. For each concentration tested, a 0. 01 ml aliquot was added to a 48-well plate containing 0.99 ml of Fluid Sabouraud Medium (DIFCO, U. S. A.) with 1 01_1 04 CFU/ml of Pityrosporum ovale (ATCC 38593). The plates were incubated at 370C for 48 hours and then 20 visually examined and scored. Vehicle-control was used as blank control. Each concentration was evaluated in duplicate. The results are shown in Table 7.

Table 7. The inhibitory effects of ginger extracts on Pityrosporum ovale (Po) MIC (Pg/ml) Ginger extract') PO Trn 1-0 100 30 I-OM 100 30 11-0 500 30 11-0c 100 30 11-OM 100 30 111-0 500 100 Blank control -- b) -- b) Prepared in Examples 1, 2 and 3 no inhibitory effect upon growth

Claims

Claims

1. A method of preparing a product potent in anti-inflammation or in anti-platelet aggregation from rhizomes of Zingiberofficinale comprising the following steps:

5 a) preparing a crude liquid from rhizomes of Zingiber officinale; b) introducing the crude liquid to a reverse phase chromatography column, and eluting the column with water, a first eluent and a second eluent in sequence, said second eluent having a polarity weaker than that of the first eluent but stronger than that of chloroform, so that a first eluate 10 resulting from elution of the first eluent and a second eluate resulting from elution of the second eluent are obtained; c) removing the first eluent from the first eluate by evaporation, so that a first concentrated eluate is obtained and is able to be used as the potent product; and 15 d) removing the second eluent from the second eluate by evaporation, so that a second concentrated eluate is obtained and is able to used as the potent product; wherein step a) comprises steps i) to iv), or comprises step 1), step I'), or step I"), wherein said steps i) to iv) are:

20 i) shedding fresh rhizomes of Zingiber officinale and filtering the resulting mixture to obtain a filtrate and a residue; ii) extracting the filtrate with a first organic solvent, recovering the resulting extraction solution of the first organic solvent, and evaporating the first organic solvent from the extraction solution to obtain a first concentrated extraction solution; iii) extracting the residue with a second organic solvent, recovering the resulting extraction solution of the second organic solvent, and 5 evaporating the second organic solvent from the extraction solution to obtain a second concentrated extraction solution; and iv) combining the first concentrated extraction solution and the second concentrated extraction solution to obtain the crude liquid; said step 1) is:

10 1) extracting powder of dried rhizomes of Zingiber officinale with the second organic solvent, recovering the resulting extraction solution of the second organic solvent, and evaporating the second organic solvent from the extraction solution to obtain the crude liquid; said step I') is:

15 1') steam distilling powder of dried rhizomes of Zingiberofficinale, and concentrating the resulting distillate by evaporation to obtain the crude liquid; and said step I") is:

I") extracting powder of dried rhizomes of Zingiber officinale with 20 supercritical C02, recovering the resulting extraction solution of the supercritical C02, and evaporating CO, from the extraction solution to obtain the crude liquid.

2. The method according to claim 1, wherein the product potent in antiinflammation or in anti-platelet aggregation comprises 0 - 10 mg 6shogaol per gram of the product, I - 150 mg 6-gingerol per gram of the product, and 0 - 40 mg 6-dehydrogingerdione per gram of the product. 5 3. The method according to claims 1 or 2, wherein said first eluent is methanol, and said second eluent is acetone 4. The method according to claims 1, 2 or 3, wherein step a) 10 comprises steps i) to iv).

5. The method according to claim 4, wherein said first organic solvent is ethyl ether.

15 6. The method according to claim 4, wherein said second organic solvent is acetone, methanol, ethanol or a combination of them.

7. The method according to claim 6, wherein said second organic solvent is acetone. 20 8. The method according to claims 1, 2 or 3, wherein step a) comprises step 1).

9. The method according to claim 8, wherein said second organic solvent is acetone, methanol, ethanol or a combination of them.

10. The method according to claim 9, wherein said second organic 5 solvent is acetone.

11. The method according to claims 1, 2 or 3, wherein step a) comprises step I').

10 12. The method according to claims 1, 2 or 3, wherein step a) comprises step I").

13. The method according to claim 1, wherein said reverse phase chromatography column is packed with a porous resin. 15 14. An anti-inflammation pharmaceutical composition comprising a therapeutically effective amount of said crude liquid prepared in step a) of the method according to claim 1, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient. 20 15. A pharmaceutical composition for the inhibition of aggregation of platelet, which comprises a therapeutically effective amount of said crude liquid prepared in step a) of the method according to claim 1, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient.

16. An anti-fungal pharmaceutical composition comprising a 5 therapeutically effective amount of said crude liquid prepared in step a) of the method according to claim 1, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient.

17. The pharmaceutical composition according to claims 14, 15 or 16, 10 wherein step a) comprises steps i) to iv).

18. The pharmaceutical composition according to claims 14, 15 or 16, wherein step a) comprises step 1).

15 19. The pharmaceutical composition according to claim 17, wherein said first organic solvent is ethyl ether.

20. The pharmaceutical composition according to claims 17, 18 or 19, wherein said second organic solvent is acetone, methanol, ethanol or a 20 combination of them.

21. The pharmaceutical composition according to claim 20, wherein said second organic solvent is acetone.

22. The pharmaceutical composition according to claims 14, 15 or 16, wherein step a) comprises step I').

23. The pharmaceutical composition according to claims 14, 15 or 16, 5 wherein step a) comprises step I").

24. An anti-inflammation pharmaceutical composition comprising a therapeutically effective amount of the product prepared according to the method of any one of claims 1 to 13, as an active ingredient, in admixture 10 with a pharmaceutically acceptable carrier or diluent for the active ingredient.

25. A pharmaceutical composition for the inhibition of aggregation of platelet, which comprises a therapeutically effective amount of the product 15 prepared according to the method of any one of claims 1-13, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient.

26. An anti-fungal pharmaceutical composition comprising a 20 therapeutically effective amount of the product prepared according to the method of any one of claims 1 to 13, as an active ingredient, in admixture with a pharmaceutically acceptable carrier or diluent for the active ingredient.

27. The pharmaceutical composition according to claims 24, 25 or 26, wherein said product is the first concentrated eluate prepared in step c).

28. The pharmaceutical composition according to claim 24, 25 or 26, 5 wherein said product is the second concentrated eluate prepared in step d).

29. The pharmaceutical composition according to claims 16 or 26, which is used in the treatment of a disease associated with Trichophyton 10 mentagrophytes or Pityrosporum ovale.

30. The pharmaceutical composition according to claim 29, in which said disease is selected from the group consisting of tinea pedis, tinea capitis, tinea cruris, tinea glabrosa, onychomycosis, pityriasis capitis, 15 pityriasis vesicolor, pityrosporurn folliculitis, seborrheic dermatitis and dandruff.

31. The pharmaceutical composition according to claim 29, which is in the form of a shampoo, a bath gel, soap, a body lotion, a body cream or a 20 detergent, 32. The pharmaceutical composition according to claim 31, which is in the form of a shampoo for use in the treatment of dandruff.