CN105510513A

CN105510513A - Method for identifying bulk pharmaceutical chemicals from ointment for promoting tissue regeneration

Info

Publication number: CN105510513A
Application number: CN201510957460.8A
Authority: CN
Inventors: 房德敏; 周永梅; 冯鑫; 徐卫国
Original assignee: TIANJIN HOSPITAL TIANJIN CITY
Current assignee: TIANJIN HOSPITAL TIANJIN CITY
Priority date: 2015-12-17
Filing date: 2015-12-17
Publication date: 2016-04-20
Anticipated expiration: 2035-12-17
Also published as: CN105510513B

Abstract

The invention provides a method for identifying bulk pharmaceutical chemicals from an ointment for promoting tissue regeneration. Thin layer chromatography is adopted for thin-layer chromatography identification of five bulk pharmaceutical chemicals of elephant hide, sesame oil, Chinese angelica, dried radix rehmanniae and crinis carbonisatus one by one from the ointment for promoting tissue regeneration. Whether ingredients of the three bulk pharmaceutical chemicals change or not is researched after the sesame oil is adopted for high-temperature frying of the Chinese angelica, the dried radix rehmanniae and the crinis carbonisatus forming the ointment for promoting tissue regeneration, and it is found that ingredients change after the sesame oil is adopted for high-temperature frying of the dried radix rehmanniae and the crinis carbonisatus, and ferulic acid contained in the Chinese angelica does not exist in the ointment after high-temperature frying is performed through the sesame oil through thin-layer chromatography identification. Substances contained inside the ointment are further determined through the method, and a great significance is achieved for follow-up research of the ointment.

Description

One carries out mirror method for distinguishing to myogenic cream Raw medicine

Technical field

The present invention relates to technical field of traditional Chinese medicines, more specifically to one, mirror method for distinguishing is carried out to myogenic cream Raw medicine.

Background technology

Treatment by Chinese herbs disease has the history of several thousand, along with the development of science and technology, Chinese medicine is also faced with modernization, go on world market, this just requires the control must carrying out traditional Chinese medicine quality, and the quality control of the component content of Chinese medicine is then most critical, especially for the Chinese medicine preparation of compound, want the quality comprehensively controlling Chinese medicine, the Accurate Measurement of the content of each composition must be carried out.

Myogenic cream comes from Zhang Shanlei " ulcer section outline ", it is TCM surgery common medicine, by more than elephant hide, calamine, tortoise plastron, blood, the medicinal material such as Radix Angelicae Sinensis, dried rhizome of rehmannia to boil through sesame oil high temperature and forms, there is myogenic, sore effect, be used for the treatment of the caused chronic wound such as wound concurrent infection, pressure ulcer, diabetes, Venous Ulcers, burn and scald evident in efficacy.Due to formulation reason, said preparation quality control lacks effective standard always, determines that this standard then needs, when measuring each component content, must to remove disturbing factor, measure content accurately, accomplishes the control drug quality of imitating, ensure the safety of medicine.

Summary of the invention

Instant invention overcomes deficiency of the prior art, provide one and mirror method for distinguishing is carried out to myogenic cream Raw medicine.

Object of the present invention is achieved by following technical proposals.

One carries out mirror method for distinguishing to myogenic cream Raw medicine, myogenic cream comprises the bulk drug of following composition by weight: elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, measure by the following method:

1. the mensuration of elephant hide:

Step 1, the preparation of elephant hide need testing solution: get myogenic cream heating water bath and make it melt, add hot water in container, fully stir, be placed in refrigerator to place, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol, ultrasonic process, filters, and obtains elephant hide need testing solution after filtrate is concentrated;

Step 2, the preparation of elephant hide control medicinal material solution: get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, in elephant hide powder, add methyl alcohol, ultrasonic process, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol residue is dissolved, obtain elephant hide control medicinal material solution;

Step 3, not containing the preparation of elephant hide negative sample solution: according to Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample heating water bath, hot water is added in container, abundant stirring, be placed in refrigerator to place, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol, ultrasonic process, filter, obtain not containing elephant hide negative sample solution after filtrate is concentrated,

Step 4, measures process: first to expansion, gets elephant hide need testing solution point in silica gel g thin-layer plate x-axis initial point place, with cyclohexane-acetone-ethyl acetate for developping agent, after launching along x-axis, taking-up is dried, and second to expansion, get elephant hide control medicinal material solution, point is about 6-8cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, after launching along the y-axis direction, taking-up is dried, and observes under being placed in ultraviolet lamp 365nm;

4. the mensuration more than blood:

Step 1, the preparation of need testing solution more than blood: get myogenic cream heating water bath and make it melt, add methyl alcohol in container, ultrasonic process, filters, and filtrate concentrates, and obtains need testing solution more than blood;

Step 2, the preparation of control medicinal material solution more than blood: get control medicinal material more than blood, remove slag with after 250 DEG C of sesame oil frying a period of times, filters, gets filtrate, in filtrate, add methyl alcohol, ultrasonic process, filters, and filtrate concentrates, and obtains control medicinal material solution more than blood;

Step 3, containing the preparation of negative sample solution more than blood: according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample heating water bath more than blood, methyl alcohol is added in container, ultrasonic process, filter, after filtrate is concentrated, obtain not containing negative sample solution more than blood,

Step 4, mensuration process: get need testing solution more than blood, containing control medicinal material solution more than negative sample solution more than blood and blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, after launching 10-12cm, taking-up is dried, then after launching twice respectively with above-mentioned developping agent, taking-up is dried, and observes under being placed in ultraviolet lamp 365nm;

5. the mensuration of the dried rhizome of rehmannia:

Step 1, the preparation of dried rhizome of rehmannia need testing solution: get myogenic cream heating water bath and make it melt, add methyl alcohol in container, ultrasonic process, filters, and filtrate is concentrated obtains dried rhizome of rehmannia need testing solution;

Step 2, the preparation of dried rhizome of rehmannia control medicinal material solution: get dried rhizome of rehmannia control medicinal material, is deep-fried with 200 DEG C of sesame oil, filters, gets filtrate, in filtrate, add methyl alcohol, ultrasonic process, filters, and after filtrate is concentrated, obtains dried rhizome of rehmannia control medicinal material solution;

Step 3, not containing the preparation of dried rhizome of rehmannia negative sample solution: according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample heating water bath, methyl alcohol is added in container, ultrasonic process, filter, filtrate is concentrated to be obtained not containing dried rhizome of rehmannia negative sample solution,

Step 4, mensuration process: get dried rhizome of rehmannia need testing solution, not containing dried rhizome of rehmannia negative sample solution and dried rhizome of rehmannia control medicinal material solution, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, after launching 15-17cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, after launching 13-15cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, after launching 13-15cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm,

4. the mensuration of Radix Angelicae Sinensis:

Step 1, the preparation of Radix Angelicae Sinensis need testing solution: get myogenic cream, adds methyl alcohol wherein, ultrasonic process, filters, and after filtrate is concentrated, obtains Radix Angelicae Sinensis need testing solution;

Step 2, the preparation of Radix Angelicae Sinensis control medicinal material solution: get control medicinal material Radix Angelicae Sinensis, adds methyl alcohol in Radix Angelicae Sinensis, ultrasonic process, filters, gets filtrate and namely obtain Radix Angelicae Sinensis control medicinal material solution;

Step 3, not containing the preparation of Radix Angelicae Sinensis negative sample solution: according to elephant hide 15-20 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample, add methyl alcohol wherein, ultrasonic process, filter, after filtrate is concentrated, obtain not containing Radix Angelicae Sinensis negative sample solution,

Step 4, mensuration process: get above-mentioned Radix Angelicae Sinensis need testing solution, not containing Radix Angelicae Sinensis negative sample solution and Radix Angelicae Sinensis control medicinal material solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm;

7. the mensuration of sesame oil:

Step 1, the preparation of sesame oil need testing solution: get myogenic cream, adds methyl alcohol wherein, ultrasonic process, filters, and after filtrate is concentrated, obtains sesame oil need testing solution;

Step 2, the preparation of sesamin reference substance solution: get sesamin reference substance, adds methyl alcohol in sesamin reference substance, makes the solution of every 1ml containing 1mg, is sesamin reference substance solution;

Step 3, not containing the preparation of sesame oil negative sample solution: according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample, add methyl alcohol wherein, ultrasonic process, filter, after filtrate is concentrated, obtain not containing sesame oil negative sample solution,

Step 4, mensuration process: get above-mentioned sesame oil need testing solution, not containing sesame oil negative sample solution and sesamin reference substance solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, take out after launching and dry, silica gel g thin-layer plate after drying sprays 10% ethanol solution of sulfuric acid, is heated to spot development clear;

8. the mensuration of forulic acid:

Step 1, the preparation of forulic acid need testing solution: get myogenic cream, heating water bath makes it melt, and adds 1% sodium bicarbonate solution wherein, ultrasonic process, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, is extracted with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol that adds makes it dissolve, and obtains forulic acid need testing solution;

Step 2, the preparation of forulic acid reference substance solution: get forulic acid reference substance, adds methyl alcohol and makes the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution in forulic acid reference substance;

Step 3, not containing the preparation of forulic acid negative sample solution: according to elephant hide 15-20 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample, heating water bath makes it melt, add 1% sodium bicarbonate solution wherein, ultrasonic process, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, extract with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol that adds makes it dissolve, obtain not containing forulic acid negative sample solution,

Step 4, mensuration process: get above-mentioned forulic acid need testing solution, forulic acid reference substance solution and do not contain forulic acid negative sample solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-dichloromethane-ethyl acetate-formic acid for developping agent, launch, take out after launching and dry, then with above-mentioned developping agent second outspread, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm.

In elephant hide measures:

In described step 1, getting myogenic cream 90-100g heating water bath makes it melt, and adds hot water 90-100mL, fully stir in container, be placed in refrigerator and place 2-4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20-30mL, ultrasonic process 20-40min, filter, filtrate obtains elephant hide need testing solution after being concentrated into 0.3-1.0mL.

In described step 2, get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, get elephant hide powder 0.5-1.5g, in elephant hide powder, add methyl alcohol 5-15mL, ultrasonic process 10-30min, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol 0.5-1.5mL makes residue dissolve, and obtains elephant hide control medicinal material solution.

In described step 3, according to Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 90-100g heating water bath, hot water 90-100mL is added in container, abundant stirring, be placed in refrigerator and place 2-4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20-30mL, ultrasonic process 20-40min, filter, filtrate obtains after being concentrated into 0.3-1.0mL not containing elephant hide negative sample solution.

In described step 4, mensuration process: first to expansion, get elephant hide need testing solution 20ul point in the silica gel g thin-layer plate x-axis initial point place of 10cm × 10cm, with cyclohexane-acetone-ethyl acetate for developping agent, its cyclohexane: acetone: the proportioning of ethyl acetate is (4-6): (1-3): (0.5-2), after launching 7.5cm along x-axis, taking-up is dried, second to expansion, get elephant hide control medicinal material solution 10ul, point is about 7cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is (4-6): (4-6): (0.1-1): (0.05-0.2), after launching along the y-axis direction, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

In the mensuration more than blood:

In described step 1, get myogenic cream 180-220g heating water bath and make it melt, add methyl alcohol 80-120mL in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL, obtains need testing solution more than blood.

In described step 2, get control medicinal material 4-6g more than blood, remove slag with after 250 DEG C of sesame oil 180-220g frying 5-15min, filter, get filtrate 15-25mL, in filtrate, add methyl alcohol 8-12mL, ultrasonic process 10-30min, filter, filtrate is concentrated into 0.5-1.5mL, obtains control medicinal material solution more than blood.

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 180-220g heating water bath more than blood, methyl alcohol 80-120mL is added in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL, obtain not containing negative sample solution more than blood.

In described step 4, mensuration process: to get more than blood need testing solution and containing each 20ul of negative sample solution more than blood, control medicinal material solution 10ul more than blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is (13-15): (3-5): (0.1-0.8), after launching 11cm, taking-up is dried, 12.5cm is launched respectively again with above-mentioned developping agent, after 14cm twice, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

In the mensuration of the dried rhizome of rehmannia:

In described step 1, get myogenic cream 180-220g heating water bath and make it melt, add methyl alcohol 80-120mL in container, ultrasonic process 20-40min, filter, filtrate obtains dried rhizome of rehmannia need testing solution after being concentrated into 0.5-1.5mL.

In described step 2, get dried rhizome of rehmannia control medicinal material 8-12g, be deep-fried with 200 DEG C of sesame oil 180-220g, filter, get filtrate 60-100mL, in filtrate, add methyl alcohol 40-80mL, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain dried rhizome of rehmannia control medicinal material solution.

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 180-220g heating water bath, methyl alcohol 80-120mL is added in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL and obtains not containing dried rhizome of rehmannia negative sample solution.

In described step 4, mensuration process: get dried rhizome of rehmannia need testing solution and do not contain each 10ul of dried rhizome of rehmannia negative sample solution, dried rhizome of rehmannia control medicinal material solution 20ul, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is (4-6): (2-4): (1-3): (0.05-0.2), after launching 16cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is (13-15): (3-5): (0.1-0.8), after launching 14cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is (4-6): (4-6): (0.1-1): (0.01-0.2), after launching 14cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

In the mensuration of Radix Angelicae Sinensis:

In described step 1, get myogenic cream 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain Radix Angelicae Sinensis need testing solution;

In described step 2, get control medicinal material Radix Angelicae Sinensis 0.3-0.8g, in Radix Angelicae Sinensis, add methyl alcohol 3-8mL, ultrasonic process 10-30min, filter, get filtrate and namely obtain Radix Angelicae Sinensis control medicinal material solution;

In described step 3, according to elephant hide 15-20 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain not containing Radix Angelicae Sinensis negative sample solution,

In described step 4, mensuration process: get above-mentioned Radix Angelicae Sinensis need testing solution and do not contain each 6ul of Radix Angelicae Sinensis negative sample solution, Radix Angelicae Sinensis control medicinal material solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm.

In the mensuration of sesame oil:

In described step 1, get myogenic cream 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain sesame oil need testing solution.

In described step 2, get sesamin reference substance 30-50mg, in sesamin reference substance, add methyl alcohol 30-50mL, make the solution of every 1ml containing 1mg, be sesamin reference substance solution.

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain not containing sesame oil negative sample solution.

In described step 4, mensuration process: get above-mentioned sesame oil need testing solution and do not contain each 10ul of sesame oil negative sample solution, sesamin reference substance solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, its cyclohexane: ether: the proportioning of ethyl acetate is (18-22): (5.0-6.0): (2.0-3.0), launch, take out after launching and dry, silica gel g thin-layer plate after drying sprays 5-15% ethanol solution of sulfuric acid, is heated to spot development clear.

In the mensuration of forulic acid:

In described step 1, get myogenic cream 8-12g, heating water bath makes it melt, and adds 1% sodium bicarbonate solution 18-22mL wherein, ultrasonic process 5-15min, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, extracts 2 times with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol 0.5-1.5mL that adds makes it dissolve, and obtains forulic acid need testing solution.

In described step 2, get forulic acid reference substance 30-50mg, in forulic acid reference substance, add methyl alcohol 30-50mL make the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution.

In described step 3, according to elephant hide 15-20 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample, 8-12g, heating water bath makes it melt, add 1% sodium bicarbonate solution 18-22mL wherein, ultrasonic process 5-15min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 0.5-1.5mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution.

In described step 4, mensuration process: get above-mentioned forulic acid need testing solution, forulic acid reference substance solution and not containing each l0ul of forulic acid negative sample solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-dichloromethane-ethyl acetate-formic acid for developping agent, its cyclohexane: methylene chloride: ethyl acetate: the proportioning of formic acid is (3-5): (0.5-2): (0.5-2): (0.05-0.2), launch, take out after launching and dry, again with above-mentioned developping agent second outspread, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm.

Beneficial effect of the present invention is: elephant hide is monarch drug in a prescription in side, and is expensive thin medicinal material, in " Chinese medicine voluminous dictionary " except proterties, does not have other specificity discrimination method.In myogenic cream, contained medicinal material is wide in variety, complicated component, elephant hide TLC distinguish once adopted multiple developping agent, one or many one-dimensional development is tested, the spot of test sample chromatogram is all smudgy clear, therefore adopts two-dimensional development method, on the basis of One step development, secondary vertically launches, and test sample chromatogram is visible clear spot on the position corresponding to control medicinal material chromatogram, Radix Angelicae Sinensis in myogenic cream, the dried rhizome of rehmannia, blood is remaining through the frying of sesame oil high temperature, Radix Angelicae Sinensis, the dried rhizome of rehmannia, with sesame oil generation chemical reaction more than blood, produce new composition, therefore be directly contrast with control medicinal material in thin layer experiment, even if adopt multiple extraction, separation method and development system test, also the object of discriminating cannot be reached, therefore with reference to control medicinal material preparation method in Asian puccoon thin-layer identification method in " Chinese Pharmacopoeia " 2010 editions one scald oil quality standard recorded, prepare more than blood according to preparation process, the fried liquid of the dried rhizome of rehmannia carries out thin layer test as the control medicinal material of TLC distinguish, in test sample chromatogram, on the position corresponding to control medicinal material chromatogram, there is the fluorescence spot of same color, more than explanation blood, the dried rhizome of rehmannia is after the frying of sesame oil high temperature, composition there occurs change, simultaneously more than blood, in the TLC distinguish of the dried rhizome of rehmannia, after One step development, thin layer blurring is overlapping, smudgy clear, degree of separation is poor, therefore employing repeatedly launches to reach separation discriminating object, TLC distinguish test has been carried out to forulic acid contained by Radix Angelicae Sinensis in myogenic cream, thin-layer developing condition differentiates forulic acid discrimination method in (3) according to the middle Radix Angelicae Sinensis of " Chinese Pharmacopoeia " 2010 editions, thin layer first time launches test sample chromatogram similar fluorescence spot on the position corresponding to reference substance chromatogram, but spot and R f value comparatively close to and spot lightness slightly difference, therefore adopt identical unfolding condition to carry out second outspread, found that spot and the reference substance chromatogram spot and R f value difference distance of test sample chromatogram increase, thus determine that the spot of test sample chromatogram and reference substance chromatogram spot are not same substances, therefore not containing forulic acid in myogenic cream.

Accompanying drawing explanation

Fig. 1 is TLC thin-layer chromatogram (under sunshine condition) in the mensuration of elephant hide in the embodiment of the present invention 4; Wherein, 1 is the elephant hide need testing solution prepared in embodiment 4, and 2 is the elephant hide control medicinal material solution prepared in embodiment 4;

Fig. 2 is TLC thin-layer chromatogram in the mensuration of elephant hide in the embodiment of the present invention 4 (under 365nm condition); Wherein, 1 is the elephant hide need testing solution prepared in embodiment 4, and 2 is the elephant hide control medicinal material solution prepared in embodiment 4;

Fig. 3 is TLC thin-layer chromatogram in the mensuration in the present invention more than blood (under 365nm condition); Wherein, be followed successively by first time expansion from left to right, second time launches and third time launches, 1 be in embodiment 1 preparation blood more than need testing solution, 2 be in embodiment 2 preparation blood more than need testing solution, 3 be in embodiment 3 preparation blood more than need testing solution, 4 be preparation in embodiment 4 blood more than control medicinal material solution, 5 be in embodiment 4 preparation do not contain negative sample solution more than blood;

Fig. 4 is TLC thin-layer chromatogram in the mensuration of the dried rhizome of rehmannia in the present invention (under 365nm condition); Wherein, 1 is the dried rhizome of rehmannia control medicinal material solution prepared in embodiment 3,2 is the dried rhizome of rehmannia control medicinal material solution prepared in embodiment 4,3 is the dried rhizome of rehmannia need testing solution prepared in embodiment 1,4 is the dried rhizome of rehmannia need testing solution prepared in embodiment 2,5 be in embodiment 3 preparation dried rhizome of rehmannia need testing solution, 6 be in embodiment 4 preparation not contain dried rhizome of rehmannia negative sample solution;

Fig. 5 is TLC thin-layer chromatogram in the mensuration of Radix Angelicae Sinensis in the present invention (under 365nm condition); Wherein, 1 be in embodiment 3 preparation not containing Radix Angelicae Sinensis negative sample solution, 2 be the Radix Angelicae Sinensis control medicinal material solution of preparation in embodiment 3, and 3 is the Radix Angelicae Sinensis need testing solution prepared in embodiment 2,4 is the Radix Angelicae Sinensis need testing solution prepared in embodiment 3, and 5 is the Radix Angelicae Sinensis need testing solution prepared in embodiment 4;

Fig. 6 is TLC thin-layer chromatogram (under sunshine condition) in the mensuration of sesame oil in the present invention; Wherein, 1 is the sesame oil need testing solution prepared in embodiment 1, and 2 is the sesame oil need testing solution prepared in embodiment 3, and 3 is the sesame oil need testing solution prepared in embodiment 4,4 be in embodiment 2 preparation sesamin reference substance solution, 5 be in embodiment 2 preparation not contain sesame oil negative sample solution;

Fig. 7 is TLC thin-layer chromatogram in the mensuration of forulic acid in the present invention; Wherein, be followed successively by first time expansion and second time expansion from left to right, 1 is the forulic acid reference substance solution prepared in embodiment 1,2 is the forulic acid need testing solution prepared in embodiment 2,3 is the forulic acid need testing solution prepared in embodiment 3,4 be in embodiment 4 preparation forulic acid need testing solution, 5 be in embodiment 1 preparation not contain forulic acid negative sample solution.

Embodiment

Below by specific embodiment, technical scheme of the present invention is further described.

The instrument adopted in experimentation and reagent, as follows:

Instrument and reagent	Producer
		TCQ-250 ultrasonic cleaner	Beijing Medical Devices two factory
SLD-I type thin-layer chromatography photographic apparatus	Tianjin Si Lida Science and Technology Ltd.
		Commercially available can cutting type silica gel g thin-layer plate	Tianjin Si Lida Science and Technology Ltd.
Cyclohexane	Tianjin Chemical Reagents Factory No.1
		Toluene	Tianjin chemical reagent wholesaling firm
Ethyl acetate	Tianjin Heng Hao company chemical reagent factory
		Acetone	Tianjin chemical reagent supply and marketing company
Sherwood oil (60-90 DEG C)	Tianjin Jin Dong Tian Zheng fine chemistry chemical reagent work
		Ether	Tianjin chemical reagent wholesaling firm
Formic acid	Tianjin Chemical Reagents Factory No.1

Embodiment 1

One carries out mirror method for distinguishing to myogenic cream Raw medicine, and myogenic cream comprises the bulk drug of following composition by weight: elephant hide 15 parts, Radix Angelicae Sinensis 10 parts, and blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, measure by the following method:

1. the mensuration of elephant hide:

One carries out mirror method for distinguishing to elephant hide in myogenic cream, and myogenic cream comprises the bulk drug of following composition by weight: elephant hide 15 parts, Radix Angelicae Sinensis 10 parts, and blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, measure by the following method:

Step 1, the preparation of elephant hide need testing solution: get myogenic cream 90g heating water bath and make it melt, add hot water 90mL in container, fully stir, be placed in refrigerator and place 2h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20mL, ultrasonic process 20min, filter, filtrate obtains elephant hide need testing solution after being concentrated into 0.3mL;

Step 2, the preparation of elephant hide control medicinal material solution: get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, get elephant hide powder 0.5g, in elephant hide powder, add methyl alcohol 5mL, ultrasonic process 10min, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol 0.5mL makes residue dissolve, and obtains elephant hide control medicinal material solution;

Step 3, not containing the preparation of elephant hide negative sample solution: according to Radix Angelicae Sinensis 10 parts, blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gesso is broken into fine powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 90g heating water bath, hot water 90mL is added in container, abundant stirring, be placed in refrigerator and place 2h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20mL, ultrasonic process 20min, filter, filtrate obtains not containing elephant hide negative sample solution after being concentrated into 0.3mL,

Step 4, mensuration process: first to expansion, get elephant hide need testing solution 20ul point in silica gel g thin-layer plate (10 × 10) x-axis initial point place, with cyclohexane-acetone-ethyl acetate for developping agent, its cyclohexane: acetone: the proportioning of ethyl acetate is 4: 3: 0.5, after launching 7.5cm along x-axis, taking-up is dried, second to expansion, get elephant hide control medicinal material solution 10ul, point is about 6cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 4: 4: 0.1: 0.05, after launching along the y-axis direction, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

2. the mensuration more than blood:

Step 1, the preparation of need testing solution more than blood: get myogenic cream 180g heating water bath and make it melt, add methyl alcohol 80mL in container, ultrasonic process 20min, filter, filtrate is concentrated into 0.5mL, obtains need testing solution more than blood;

Step 2, the preparation of control medicinal material solution more than blood: get control medicinal material 4g more than blood, remove slag with after 250 DEG C of sesame oil 180g frying 5min, filters, get filtrate 15mL, in filtrate, add methyl alcohol 8mL, ultrasonic process 10min, filter, filtrate is concentrated into 0.5mL, obtains control medicinal material solution more than blood;

Step 3, containing the preparation of negative sample solution more than blood: according to elephant hide 15 parts, Radix Angelicae Sinensis 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 180g heating water bath more than blood, methyl alcohol 80mL is added in container, ultrasonic process 20min, filter, filtrate is concentrated into 0.5mL, obtain not containing negative sample solution more than blood,

Step 4, mensuration process: to get more than blood need testing solution and containing control medicinal material solution 10ul more than each 20ul of negative sample solution, blood more than blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 13: 3: 0.1, after launching 10cm, taking-up is dried, after launching 12.5cm, 14cm twice respectively with above-mentioned developping agent again, taking-up is dried, and observes under being placed in ultraviolet lamp 365nm.

3. the mensuration of the dried rhizome of rehmannia:

Step 1, the preparation of dried rhizome of rehmannia need testing solution: get myogenic cream 180g heating water bath and make it melt, add methyl alcohol 80mL in container, ultrasonic process 20min, filter, filtrate obtains dried rhizome of rehmannia need testing solution after being concentrated into 0.5mL;

Step 2, the preparation of dried rhizome of rehmannia control medicinal material solution: get dried rhizome of rehmannia control medicinal material 8g, is deep-fried with 200 DEG C of sesame oil 180g, filters, get filtrate 60mL, in filtrate, add methyl alcohol 40mL, ultrasonic process 10min, filter, after filtrate is concentrated into 0.5mL, obtain dried rhizome of rehmannia control medicinal material solution;

Step 3, not containing the preparation of dried rhizome of rehmannia negative sample solution: according to elephant hide 15 parts, Radix Angelicae Sinensis 10 parts, blood mores than 10 parts, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 180g heating water bath, methyl alcohol 80mL is added in container, ultrasonic process 20min, filter, filtrate is concentrated into 0.5mL and obtains not containing dried rhizome of rehmannia negative sample solution,

Step 4, mensuration process: get dried rhizome of rehmannia need testing solution and do not contain each 10ul of dried rhizome of rehmannia negative sample solution, dried rhizome of rehmannia control medicinal material solution 20ul, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 4: 2: 1: 0.05, after launching 15cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 15: 5: 0.8, after launching 13cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 6: 6: 1: 0.2, after launching 13cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

4. the mensuration of Radix Angelicae Sinensis:

Step 1, the preparation of Radix Angelicae Sinensis need testing solution: get myogenic cream 8g, adds methyl alcohol 18mL wherein, ultrasonic process 10min, filters, after filtrate is concentrated into 0.5mL, obtains Radix Angelicae Sinensis need testing solution;

Step 2, the preparation of Radix Angelicae Sinensis control medicinal material solution: get control medicinal material Radix Angelicae Sinensis 0.3g, add methyl alcohol 3mL in Radix Angelicae Sinensis, ultrasonic process 10min, filters, gets filtrate and namely obtain Radix Angelicae Sinensis control medicinal material solution;

Step 3, not containing the preparation of Radix Angelicae Sinensis negative sample solution: according to elephant hide 15 parts, blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample 8g, add methyl alcohol 18mL wherein, ultrasonic process 10min, filter, after filtrate is concentrated into 0.5mL, obtain not containing Radix Angelicae Sinensis negative sample solution,

Step 4, mensuration process: get above-mentioned Radix Angelicae Sinensis need testing solution and do not contain each 6ul of Radix Angelicae Sinensis negative sample solution, Radix Angelicae Sinensis control medicinal material solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm;

5. the mensuration of sesame oil:

Step 1, the preparation of sesame oil need testing solution: get myogenic cream 8g, adds methyl alcohol 18mL wherein, ultrasonic process 10min, filters, after filtrate is concentrated into 0.5mL, obtains sesame oil need testing solution;

Step 2, the preparation of sesamin reference substance solution: get sesamin reference substance 30mg, adds methyl alcohol 30mL in sesamin reference substance, makes the solution of every 1ml containing 1mg, is sesamin reference substance solution;

Step 3, not containing the preparation of sesame oil negative sample solution: according to elephant hide 15 parts, Radix Angelicae Sinensis 10 parts, blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 8g, add methyl alcohol 18mL wherein, ultrasonic process 10min, filter, after filtrate is concentrated into 0.5mL, obtain not containing sesame oil negative sample solution,

Step 4, mensuration process: get above-mentioned sesame oil need testing solution and do not contain each 10ul of sesame oil negative sample solution, sesamin reference substance solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, its cyclohexane: ether: the proportioning of ethyl acetate is 18: 5.0: 2.0, launches, takes out after launching and dries, silica gel g thin-layer plate after drying sprays 15% ethanol solution of sulfuric acid, is heated to spot development clear.

6. the mensuration of forulic acid:

Step 1, the preparation of forulic acid need testing solution: get myogenic cream 8g, heating water bath makes it melt, and adds 1% sodium bicarbonate solution 18mL wherein, ultrasonic process 5min, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, extracts 2 times with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol 0.5mL that adds makes it dissolve, and obtains forulic acid need testing solution;

Step 2, the preparation of forulic acid reference substance solution: get forulic acid reference substance 30mg, adds methyl alcohol 30mL and makes the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution in forulic acid reference substance;

Step 3, not containing the preparation of forulic acid negative sample solution: according to elephant hide 15 parts, blood mores than 10 parts, 20 parts, the dried rhizome of rehmannia, tortoise plastron 20 parts, 400 parts, sesame oil, calamine 28 parts, plaster stone 25 parts and 80 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample, 8g, heating water bath makes it melt, add 1% sodium bicarbonate solution 18mL wherein, ultrasonic process 5min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 0.5mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution,

Step 4, mensuration process: get above-mentioned forulic acid need testing solution, forulic acid reference substance solution and do not contain each l0ul of forulic acid negative sample solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-dichloromethane-ethyl acetate-formic acid for developping agent, its cyclohexane: methylene chloride: ethyl acetate: the proportioning of formic acid is 5: 2: 2: 0.2, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm.

Embodiment 2

One carries out mirror method for distinguishing to myogenic cream Raw medicine, and myogenic cream comprises the bulk drug of following composition by weight: elephant hide 20 parts, Radix Angelicae Sinensis 15 parts, and blood mores than 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, measure by the following method:

1. the mensuration of elephant hide:

Step 1, the preparation of elephant hide need testing solution: get myogenic cream 100g heating water bath and make it melt, add hot water 100mL in container, fully stir, be placed in refrigerator and place 4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 30mL, ultrasonic process 40min, filter, filtrate obtains elephant hide need testing solution after being concentrated into 1.0mL;

Step 2, the preparation of elephant hide control medicinal material solution: get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, get elephant hide powder 1.5g, in elephant hide, add methyl alcohol 15mL, ultrasonic process 30min, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol 1.5mL makes residue dissolve, and obtains elephant hide control medicinal material solution;

Step 3, not containing the preparation of elephant hide negative sample solution: according to Radix Angelicae Sinensis 15 parts, blood mores than 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gesso is broken into impalpable powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 100g heating water bath, hot water 100mL is added in container, abundant stirring, be placed in refrigerator and place 4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 30mL, ultrasonic process 40min, filter, filtrate obtains not containing elephant hide negative sample solution after being concentrated into 1.0mL,

Step 4, mensuration process: first to expansion, get elephant hide need testing solution 20ul point in silica gel g thin-layer plate (10 × 10) x-axis initial point place, with cyclohexane-acetone-ethyl acetate for developping agent, its cyclohexane: acetone: the proportioning of ethyl acetate is 6: 3: 2, after launching 7.5cm along x-axis, taking-up is dried, second to expansion, get elephant hide control medicinal material solution 10ul, point is about 8cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 6: 6: 1: 0.2, after launching along the y-axis direction, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

2. the mensuration more than blood:

Step 1, the preparation of need testing solution more than blood: get myogenic cream 220g heating water bath and make it melt, add methyl alcohol 120mL in container, ultrasonic process 40min, filter, filtrate is concentrated into 1.5mL, obtains need testing solution more than blood;

Step 2, the preparation of control medicinal material solution more than blood: get control medicinal material 6g more than blood, remove slag with after 250 DEG C of sesame oil 220g frying 15min, filters, get filtrate 25mL, in filtrate, add methyl alcohol 12mL, ultrasonic process 30min, filter, filtrate is concentrated into 1.5mL, obtains control medicinal material solution more than blood;

Step 3, containing the preparation of negative sample solution more than blood: according to elephant hide 20 parts, Radix Angelicae Sinensis 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 220g heating water bath more than blood, methyl alcohol 120mL is added in container, ultrasonic process 40min, filter, filtrate is concentrated into 1.5mL, obtain not containing negative sample solution more than blood,

Step 4, mensuration process: to get more than blood need testing solution and containing control medicinal material solution 10ul more than each 20ul of negative sample solution, blood more than blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 15: 5: 0.8, after launching 12cm, taking-up is dried, after launching 12.5cm, 14cm twice respectively with above-mentioned developping agent again, taking-up is dried, and observes under being placed in ultraviolet lamp 365nm.

3. the mensuration of the dried rhizome of rehmannia:

Step 1, the preparation of dried rhizome of rehmannia need testing solution: get myogenic cream 220g heating water bath and make it melt, add methyl alcohol 120mL in container, ultrasonic process 40min, filter, filtrate obtains dried rhizome of rehmannia need testing solution after being concentrated into 1.5mL;

Step 2, the preparation of dried rhizome of rehmannia control medicinal material solution: get dried rhizome of rehmannia control medicinal material 12g, is deep-fried with 200 DEG C of sesame oil 220g, filters, get filtrate 100mL, in filtrate, add methyl alcohol 80mL, ultrasonic process 30min, filter, after filtrate is concentrated into 1.5mL, obtain dried rhizome of rehmannia control medicinal material solution;

Step 3, not containing the preparation of dried rhizome of rehmannia negative sample solution: according to elephant hide 20 parts, Radix Angelicae Sinensis 15 parts, blood mores than 15 parts, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 220g heating water bath, methyl alcohol 120mL is added in container, ultrasonic process 40min, filter, filtrate is concentrated into 1.5mL and obtains not containing dried rhizome of rehmannia negative sample solution,

Step 4, mensuration process: get dried rhizome of rehmannia need testing solution and do not contain each 10ul of dried rhizome of rehmannia negative sample solution, dried rhizome of rehmannia control medicinal material solution 20ul, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 6: 4: 3: 0.2, after launching 17cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 13: 3: 0.1, after launching 15cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 4: 4: 0.1: 0.01, after launching 15cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm.

4. the mensuration of Radix Angelicae Sinensis:

Step 1, the preparation of Radix Angelicae Sinensis need testing solution: get myogenic cream 12g, adds methyl alcohol 22mL wherein, ultrasonic process 30min, filters, after filtrate is concentrated into 1.5mL, obtains Radix Angelicae Sinensis need testing solution;

Step 2, the preparation of Radix Angelicae Sinensis control medicinal material solution: get control medicinal material Radix Angelicae Sinensis 0.8g, add methyl alcohol 8mL in Radix Angelicae Sinensis, ultrasonic process 30min, filters, gets filtrate and namely obtain Radix Angelicae Sinensis control medicinal material solution;

Step 3, not containing the preparation of Radix Angelicae Sinensis negative sample solution: according to elephant hide 20 parts, blood mores than 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample 12g, add methyl alcohol 22mL wherein, ultrasonic process 30min, filter, after filtrate is concentrated into 1.5mL, obtain not containing Radix Angelicae Sinensis negative sample solution,

5. the mensuration of sesame oil:

Step 1, the preparation of sesame oil need testing solution: get myogenic cream 12g, adds methyl alcohol 22mL wherein, ultrasonic process 30min, filters, after filtrate is concentrated into 1.5mL, obtains sesame oil need testing solution;

Step 2, the preparation of sesamin reference substance solution: get sesamin reference substance 50mg, adds methyl alcohol 50mL in sesamin reference substance, makes the solution of every 1ml containing 1mg, is sesamin reference substance solution;

Step 3, not containing the preparation of sesame oil negative sample solution: according to elephant hide 20 parts, Radix Angelicae Sinensis 15 parts, blood mores than 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 12g, add methyl alcohol 22mL wherein, ultrasonic process 30min, filter, after filtrate is concentrated into 1.5mL, obtain not containing sesame oil negative sample solution,

Step 4, mensuration process: get above-mentioned sesame oil need testing solution and do not contain each 10ul of sesame oil negative sample solution, sesamin reference substance solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, its cyclohexane: ether: the proportioning of ethyl acetate is 22: 6.0: 3.0, launches, takes out after launching and dries, silica gel g thin-layer plate after drying sprays 5% ethanol solution of sulfuric acid, is heated to spot development clear.

6. the mensuration of forulic acid:

Step 1, the preparation of forulic acid need testing solution: get myogenic cream 12g, heating water bath makes it melt, and adds 1% sodium bicarbonate solution 22mL wherein, ultrasonic process 15min, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, extracts 2 times with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol 1.5mL that adds makes it dissolve, and obtains forulic acid need testing solution;

Step 2, the preparation of forulic acid reference substance solution: get forulic acid reference substance 50mg, adds methyl alcohol 50mL and makes the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution in forulic acid reference substance;

Step 3, not containing the preparation of forulic acid negative sample solution: according to elephant hide 20 parts, blood mores than 15 parts, 30 parts, the dried rhizome of rehmannia, tortoise plastron 30 parts, 500 parts, sesame oil, calamine 32 parts, plaster stone 30 parts and 100 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample 12g, heating water bath makes it melt, add 1% sodium bicarbonate solution 22mL wherein, ultrasonic process 15min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 1.5mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution,

Step 4, mensuration process: get above-mentioned forulic acid need testing solution, forulic acid reference substance solution and do not contain each l0ul of forulic acid negative sample solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-dichloromethane-ethyl acetate-formic acid for developping agent, its cyclohexane: methylene chloride: ethyl acetate: the proportioning of formic acid is 3: 0.5: 0.5: 0.05, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm.

Embodiment 3

One carries out mirror method for distinguishing to myogenic cream Raw medicine, and myogenic cream comprises the bulk drug of following composition by weight: elephant hide 16 parts, Radix Angelicae Sinensis 11 parts, and blood mores than 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, measure by the following method:

1. the mensuration of elephant hide:

Step 1, the preparation of elephant hide need testing solution: get myogenic cream 100g heating water bath and make it melt, add hot water 100mL in container, fully stir, be placed in refrigerator and place 3h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 25mL, ultrasonic process 30min, filter, filtrate obtains elephant hide need testing solution after being concentrated into 0.5mL;

Step 2, the preparation of elephant hide control medicinal material solution: get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, get elephant hide powder 1g, in elephant hide, add methyl alcohol 10mL, ultrasonic process 20min, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol 1mL makes residue dissolve, and obtains elephant hide control medicinal material solution;

Step 3, not containing the preparation of elephant hide negative sample solution: according to Radix Angelicae Sinensis 11 parts, blood mores than 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gesso is broken into impalpable powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 100g heating water bath, hot water 100mL is added in container, abundant stirring, be placed in refrigerator and place 3h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 25mL, ultrasonic process 30min, filter, filtrate obtains not containing elephant hide negative sample solution after being concentrated into 0.5mL,

Step 4, mensuration process: first to expansion, get elephant hide need testing solution 20ul point in silica gel g thin-layer plate (10 × 10) x-axis initial point place, with cyclohexane-acetone-ethyl acetate for developping agent, its cyclohexane: acetone: the proportioning of ethyl acetate is 5: 2: 1, after launching 7.5cm along x-axis, taking-up is dried, second to expansion, get elephant hide control medicinal material solution 10ul, point is about 7cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, after launching along the y-axis direction, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, in elephant hide test sample chromatogram, on the position corresponding to control medicinal material elephant hide chromatogram, the fluorescence spot of aobvious same color, not noiseless containing the contrast of elephant hide negative sample.

2. the mensuration more than blood:

Step 1, the preparation of need testing solution more than blood: get myogenic cream 200g heating water bath and make it melt, add methyl alcohol 100mL in container, ultrasonic process 20min, filter, filtrate is concentrated into 1mL, obtains need testing solution more than blood;

Step 2, the preparation of control medicinal material solution more than blood: get control medicinal material 5g more than blood, remove slag with after 250 DEG C of sesame oil 200g frying 10min, filters, get filtrate 20mL, in filtrate, add methyl alcohol 10mL, ultrasonic process 20min, filter, filtrate is concentrated into 1mL, obtains control medicinal material solution more than blood;

Step 3, containing the preparation of negative sample solution more than blood: according to elephant hide 16 parts, Radix Angelicae Sinensis 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 200g heating water bath more than blood, methyl alcohol 100mL is added in container, ultrasonic process 20min, filter, filtrate is concentrated into 1mL, obtain not containing negative sample solution more than blood,

Step 4, mensuration process: to get more than blood need testing solution and containing each 20ul of negative sample solution more than blood, control medicinal material solution 10ul more than blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 14: 4: 0.4, after launching 11cm, taking-up is dried, 12.5cm is launched respectively again with above-mentioned developping agent, after 14cm twice, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, more than blood in test sample chromatogram, to blood more than on the corresponding position of control medicinal material chromatogram, the fluorescence spot of aobvious same color, noiseless containing negative sample contrast more than blood.

3. the mensuration of the dried rhizome of rehmannia:

Step 1, the preparation of dried rhizome of rehmannia need testing solution: get myogenic cream 200g heating water bath and make it melt, add methyl alcohol 100mL in container, ultrasonic process 30min, filter, filtrate obtains dried rhizome of rehmannia need testing solution after being concentrated into 1mL;

Step 2, the preparation of dried rhizome of rehmannia control medicinal material solution: get dried rhizome of rehmannia control medicinal material 10g, is deep-fried with 200 DEG C of sesame oil 200g, filters, get filtrate 80mL, in filtrate, add methyl alcohol 60mL, ultrasonic process 20min, filter, after filtrate is concentrated into 1mL, obtain dried rhizome of rehmannia control medicinal material solution;

Step 3, not containing the preparation of dried rhizome of rehmannia negative sample solution: according to elephant hide 16 parts, Radix Angelicae Sinensis 11 parts, blood mores than 11 parts, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 200g heating water bath, methyl alcohol 100mL is added in container, ultrasonic process 30min, filter, filtrate obtains not containing dried rhizome of rehmannia negative sample solution after being concentrated into 1mL,

Step 4, mensuration process: get dried rhizome of rehmannia need testing solution and do not contain each 10ul of dried rhizome of rehmannia negative sample solution, dried rhizome of rehmannia control medicinal material solution 20ul, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 3: 2: 0.1, after launching 16cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 14: 4: 0.4, after launching 14cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, after launching 14cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, in dried rhizome of rehmannia test sample chromatogram, on the position corresponding to dried rhizome of rehmannia control medicinal material chromatogram, the fluorescence spot of aobvious same color, not noiseless containing the contrast of dried rhizome of rehmannia negative sample.

4. the mensuration of Radix Angelicae Sinensis:

Step 1, the preparation of Radix Angelicae Sinensis need testing solution: get myogenic cream 10g, adds methyl alcohol 20mL wherein, ultrasonic process 20min, filters, after filtrate is concentrated into 1mL, obtains Radix Angelicae Sinensis need testing solution;

Step 2, the preparation of Radix Angelicae Sinensis control medicinal material solution: get control medicinal material Radix Angelicae Sinensis 0.5g, add methyl alcohol 5mL in Radix Angelicae Sinensis, ultrasonic process 20min, filters, gets filtrate and namely obtain Radix Angelicae Sinensis control medicinal material solution;

Step 3, not containing the preparation of Radix Angelicae Sinensis negative sample solution: according to elephant hide 16 parts, blood mores than 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample 10g, add methyl alcohol 20mL wherein, ultrasonic process 20min, filter, after filtrate is concentrated into 1mL, obtain not containing Radix Angelicae Sinensis negative sample solution,

Step 4, mensuration process: get above-mentioned Radix Angelicae Sinensis need testing solution and do not contain each 6ul of Radix Angelicae Sinensis negative sample solution, Radix Angelicae Sinensis control medicinal material solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm, in Radix Angelicae Sinensis test sample chromatogram, on the position corresponding to control medicinal material Radix Angelicae Sinensis chromatogram, the fluorescence spot of aobvious same color, not noiseless containing the contrast of Radix Angelicae Sinensis negative sample,

5. the mensuration of sesame oil:

Step 1, the preparation of sesame oil need testing solution: get myogenic cream 10g, adds methyl alcohol 20mL wherein, ultrasonic process 20min, filters, after filtrate is concentrated into 1mL, obtains sesame oil need testing solution;

Step 2, the preparation of sesamin reference substance solution: get sesamin reference substance 40mg, adds methyl alcohol 40mL in sesamin reference substance, makes the solution of every 1ml containing 1mg, is sesamin reference substance solution;

Step 3, not containing the preparation of sesame oil negative sample solution: according to elephant hide 16 parts, Radix Angelicae Sinensis 11 parts, blood mores than 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 10g, add methyl alcohol 20mL wherein, ultrasonic process 20min, filter, after filtrate is concentrated into 1mL, obtain not containing sesame oil negative sample solution,

Step 4, mensuration process: get above-mentioned sesame oil need testing solution and do not contain each 10ul of sesame oil negative sample solution, sesamin reference substance solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, its cyclohexane: ether: the proportioning of ethyl acetate is 20: 5.0: 3.0, launch, take out after launching and dry, silica gel g thin-layer plate after drying sprays 12% ethanol solution of sulfuric acid, be heated to spot development clear, in sesame oil test sample chromatogram, on the position corresponding to reference substance sesame oil chromatogram, the spot of aobvious same color, not noiseless containing the contrast of sesame oil negative sample.

6. the mensuration of forulic acid:

Step 1, the preparation of forulic acid need testing solution: get myogenic cream 10g, heating water bath makes it melt, and adds 1% sodium bicarbonate solution 20mL wherein, ultrasonic process 10min, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, extracts 2 times with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol 1mL that adds makes it dissolve, and obtains forulic acid need testing solution;

Step 2, the preparation of forulic acid reference substance solution: get forulic acid reference substance 40mg, adds methyl alcohol 40mL and makes the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution in forulic acid reference substance;

Step 3, not containing the preparation of forulic acid negative sample solution: according to elephant hide 16 parts, blood mores than 11 parts, 22 parts, the dried rhizome of rehmannia, tortoise plastron 22 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 26 parts and 85 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample 10g, heating water bath makes it melt, add 1% sodium bicarbonate solution 20mL wherein, ultrasonic process 10min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 1mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution,

Step 4, mensuration process: get above-mentioned forulic acid need testing solution, forulic acid reference substance solution and do not contain each l0ul of forulic acid negative sample solution, put respectively on same silica gel g thin-layer plate, with cyclohexane-dichloromethane-ethyl acetate-formic acid for developping agent, its cyclohexane: methylene chloride: ethyl acetate: the proportioning of formic acid is 4: 1: 1: 0.1, launch, take out after launching and dry, observe under being placed in ultraviolet lamp 365nm, in forulic acid test sample chromatogram, on the position corresponding to reference substance forulic acid chromatogram, without the fluorescence spot of aobvious same color.

Embodiment 4

One carries out mirror method for distinguishing to myogenic cream Raw medicine, and myogenic cream comprises the bulk drug of following composition by weight: elephant hide 19 parts, Radix Angelicae Sinensis 13 parts, and blood mores than 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, measure by the following method:

1. the mensuration of elephant hide:

Step 3, not containing the preparation of elephant hide negative sample solution: according to Radix Angelicae Sinensis 13 parts, blood mores than 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gesso is broken into impalpable powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 100g heating water bath, hot water 100mL is added in container, abundant stirring, be placed in refrigerator and place 3h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 25mL, ultrasonic process 30min, filter, filtrate obtains not containing elephant hide negative sample solution after being concentrated into 0.5mL,

Step 4, mensuration process: first to expansion, get elephant hide need testing solution 20ul point in silica gel g thin-layer plate (10 × 10) x-axis initial point place, with cyclohexane-acetone-ethyl acetate for developping agent, its cyclohexane: acetone: the proportioning of ethyl acetate is 5: 2: 1, after launching 7.5cm along x-axis, taking-up is dried, second to expansion, get elephant hide control medicinal material solution 10ul, point is about 6.5cm place in x-axis apart from initial point, with cyclohexane-toluene-ethyl acetate-formic acid for developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, after launching along the y-axis direction, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, in elephant hide test sample chromatogram, on the position corresponding to control medicinal material elephant hide chromatogram, the fluorescence spot of aobvious same color, not noiseless containing the contrast of elephant hide negative sample.

2. the mensuration more than blood:

Step 3, containing the preparation of negative sample solution more than blood: according to elephant hide 19 parts, Radix Angelicae Sinensis 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 200g heating water bath more than blood, methyl alcohol 100mL is added in container, ultrasonic process 20min, filter, filtrate is concentrated into 1mL, obtain not containing negative sample solution more than blood,

Step 4, mensuration process: to get more than blood need testing solution and containing each 20ul of negative sample solution more than blood, control medicinal material solution 10ul more than blood, put respectively on the thin layer plate of same silica G, with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid for developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 14: 4: 0.4, after launching 10.5cm, taking-up is dried, 12.5cm is launched respectively again with above-mentioned developping agent, after 14cm twice, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, more than blood in test sample chromatogram, to blood more than on the corresponding position of control medicinal material chromatogram, the fluorescence spot of aobvious same color, noiseless containing negative sample contrast more than blood.

3. the mensuration of the dried rhizome of rehmannia:

Step 3, not containing the preparation of dried rhizome of rehmannia negative sample solution: according to elephant hide 19 parts, Radix Angelicae Sinensis 13 parts, blood mores than 13 parts, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 200g heating water bath, methyl alcohol 100mL is added in container, ultrasonic process 30min, filter, filtrate obtains not containing dried rhizome of rehmannia negative sample solution after being concentrated into 1mL,

Step 4, mensuration process: get dried rhizome of rehmannia need testing solution and do not contain each 10ul of dried rhizome of rehmannia negative sample solution, dried rhizome of rehmannia control medicinal material solution 20ul, put respectively on the thin layer plate of same silica G, first time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 3: 2: 0.1, after launching 15.5cm, taking-up is dried, second time is launched with sherwood oil (60-90 DEG C)-acetic ether-methanoic acid as developping agent, wherein sherwood oil (60-90 DEG C): ethyl acetate: the proportioning of formic acid is 14: 4: 0.4, after launching 13.5cm, taking-up is dried, third time launches with cyclohexane-toluene-ethyl acetate-formic acid as developping agent, its cyclohexane: toluene: ethyl acetate: the proportioning of formic acid is 5: 5: 0.5: 0.1, after launching 13.5cm, taking-up is dried, observe under being placed in ultraviolet lamp 365nm, in dried rhizome of rehmannia test sample chromatogram, on the position corresponding to dried rhizome of rehmannia control medicinal material chromatogram, the fluorescence spot of aobvious same color, not noiseless containing the contrast of dried rhizome of rehmannia negative sample.

4. the mensuration of Radix Angelicae Sinensis:

Step 3, not containing the preparation of Radix Angelicae Sinensis negative sample solution: according to elephant hide 19 parts, blood mores than 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gypsum, elephant hide is ground into impalpable powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing Radix Angelicae Sinensis negative sample, get not containing Radix Angelicae Sinensis negative sample 10g, add methyl alcohol 20mL wherein, ultrasonic process 20min, filter, after filtrate is concentrated into 1mL, obtain not containing Radix Angelicae Sinensis negative sample solution,

5. the mensuration of sesame oil:

Step 3, not containing the preparation of sesame oil negative sample solution: according to elephant hide 19 parts, Radix Angelicae Sinensis 13 parts, blood mores than 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 10g, add methyl alcohol 20mL wherein, ultrasonic process 20min, filter, after filtrate is concentrated into 1mL, obtain not containing sesame oil negative sample solution,

Step 4, mensuration process: get above-mentioned sesame oil need testing solution and do not contain each 10ul of sesame oil negative sample solution, sesamin reference substance solution 5ul, put respectively on same silica gel g thin-layer plate, with cyclohexane-ether-ethyl acetate for developping agent, its cyclohexane: ether: the proportioning of ethyl acetate is 20: 5.5: 2.5, launch, take out after launching and dry, silica gel g thin-layer plate after drying sprays 10% ethanol solution of sulfuric acid, be heated to spot development clear, in sesame oil test sample chromatogram, on the position corresponding to reference substance sesame oil chromatogram, the spot of aobvious same color, not noiseless containing the contrast of sesame oil negative sample.

6. the mensuration of forulic acid:

Step 3, not containing the preparation of forulic acid negative sample solution: according to elephant hide 19 parts, blood mores than 13 parts, 25 parts, the dried rhizome of rehmannia, tortoise plastron 25 parts, 450 parts, sesame oil, calamine 29 parts, plaster stone 29 parts and 88 parts, beeswax, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample 10g, heating water bath makes it melt, add 1% sodium bicarbonate solution 20mL wherein, ultrasonic process 10min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 1mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution,

Above to invention has been exemplary description; should be noted that; when not departing from core of the present invention, any simple distortion, amendment or other those skilled in the art can not spend the equivalent replacement of creative work all to fall into protection scope of the present invention.

Claims

1. one kind is carried out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: myogenic cream comprises the bulk drug of following composition by weight: elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, measure by the following method:

1). the mensuration of elephant hide:

2). the mensuration more than blood:

3). the mensuration of the dried rhizome of rehmannia:

4). the mensuration of Radix Angelicae Sinensis:

5). the mensuration of sesame oil:

6). the mensuration of forulic acid:

2. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in elephant hide measures:

In described step 1, getting myogenic cream 90-100g heating water bath makes it melt, and adds hot water 90-100mL, fully stir in container, be placed in refrigerator and place 2-4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20-30mL, ultrasonic process 20-40min, filter, filtrate obtains elephant hide need testing solution after being concentrated into 0.3-1.0mL;

In described step 2, get elephant hide control medicinal material talcum powder scald to foaming after be ground into fine powder, obtain elephant hide powder, get elephant hide powder 0.5-1.5g, in elephant hide powder, add methyl alcohol 5-15mL, ultrasonic process 10-30min, filter, get filtrate, obtain residue by after filtrate evaporate to dryness, in residue, add methyl alcohol 0.5-1.5mL makes residue dissolve, and obtains elephant hide control medicinal material solution;

In described step 3, according to Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone and gesso, constantly be stirred to and be cooled to cream, obtain not containing elephant hide negative sample, get and do not make it melt containing elephant hide negative sample 90-100g heating water bath, hot water 90-100mL is added in container, abundant stirring, be placed in refrigerator and place 2-4h, after upper strata is solidified, take off a layer suspension, filter, residue adds methyl alcohol 20-30mL, ultrasonic process 20-40min, filter, filtrate obtains not containing elephant hide negative sample solution after being concentrated into 0.3-1.0mL,

3. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in the mensuration more than blood:

In described step 1, get myogenic cream 180-220g heating water bath and make it melt, add methyl alcohol 80-120mL in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL, obtains need testing solution more than blood;

In described step 2, get control medicinal material 4-6g more than blood, remove slag with after 250 DEG C of sesame oil 180-220g frying 5-15min, filter, get filtrate 15-25mL, in filtrate, add methyl alcohol 8-12mL, ultrasonic process 10-30min, filter, filtrate is concentrated into 0.5-1.5mL, obtains control medicinal material solution more than blood;

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing negative sample more than blood, get and do not make it melt containing negative sample 180-220g heating water bath more than blood, methyl alcohol 80-120mL is added in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL, obtain not containing negative sample solution more than blood,

4. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in the mensuration of the dried rhizome of rehmannia:

In described step 1, get myogenic cream 180-220g heating water bath and make it melt, add methyl alcohol 80-120mL in container, ultrasonic process 20-40min, filter, filtrate obtains dried rhizome of rehmannia need testing solution after being concentrated into 0.5-1.5mL;

In described step 2, get dried rhizome of rehmannia control medicinal material 8-12g, be deep-fried with 200 DEG C of sesame oil 180-220g, filter, get filtrate 60-100mL, in filtrate, add methyl alcohol 40-80mL, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain dried rhizome of rehmannia control medicinal material solution;

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, Radix Angelicae Sinensis, tortoise plastron, blood is remaining fried withered with putting in pot with sesame oil, remove slag, filter, filter after beeswax is melted completely in deep fat, lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing dried rhizome of rehmannia negative sample, get and do not make it melt containing dried rhizome of rehmannia negative sample 180-220g heating water bath, methyl alcohol 80-120mL is added in container, ultrasonic process 20-40min, filter, filtrate is concentrated into 0.5-1.5mL and obtains not containing dried rhizome of rehmannia negative sample solution,

5. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in the mensuration of Radix Angelicae Sinensis:

6. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in the mensuration of sesame oil:

In described step 1, get myogenic cream 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain sesame oil need testing solution;

In described step 2, get sesamin reference substance 30-50mg, in sesamin reference substance, add methyl alcohol 30-50mL, make the solution of every 1ml containing 1mg, be sesamin reference substance solution;

In described step 3, according to elephant hide 15-20 part, Radix Angelicae Sinensis 10-15 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, obtain elephant hide powder, sieving for standby, above-mentioned raw materials is mixed, obtain not containing sesame oil negative sample, get not containing sesame oil negative sample 8-12g, add methyl alcohol 18-22mL wherein, ultrasonic process 10-30min, filter, after filtrate is concentrated into 0.5-1.5mL, obtain not containing sesame oil negative sample solution,

7. one according to claim 1 carries out mirror method for distinguishing to myogenic cream Raw medicine, it is characterized in that: in the mensuration of forulic acid:

In described step 1, get myogenic cream 8-12g, heating water bath makes it melt, and adds 1% sodium bicarbonate solution 18-22mL wherein, ultrasonic process 5-15min, filter, gained filtrate, with watery hydrochloric acid adjust ph to 2 ~ 3, extracts 2 times with ether jolting, merge extract, volatilized by ether, the residue obtained methyl alcohol 0.5-1.5mL that adds makes it dissolve, and obtains forulic acid need testing solution;

In described step 2, get forulic acid reference substance 30-50mg, in forulic acid reference substance, add methyl alcohol 30-50mL make the solution of every 1ml containing 1mg, obtain forulic acid reference substance solution;

In described step 3, according to elephant hide 15-20 part, 10-15 part more than blood, dried rhizome of rehmannia 20-30 part, tortoise plastron 20-30 part, sesame oil 400-500 part, calamine 28-32 part, plaster stone 25-30 part and beeswax 80-100 part, by calamine, gesso is broken into fine powder, elephant hide talcum powder scalds and be ground into fine powder to foaming, sieving for standby, tortoise plastron, more than blood, the dried rhizome of rehmannia and sesame oil are put together in pot fried withered, remove slag, filter, filter after beeswax is melted completely in deep fat and lower the temperature from fire, add calamine stone, gesso and elephant hide powder, constantly be stirred to and be cooled to cream, obtain not containing forulic acid negative sample, get not containing forulic acid negative sample, 8-12g, heating water bath makes it melt, add 1% sodium bicarbonate solution 18-22mL wherein, ultrasonic process 5-15min, filter, gained filtrate is with watery hydrochloric acid adjust ph to 2 ~ 3, 2 times are extracted with ether jolting, merge extract, ether is volatilized, the residue obtained methyl alcohol 0.5-1.5mL that adds makes it dissolve, obtain not containing forulic acid negative sample solution,