CN1175877C - Fatty emulsion of goldenrod for injection and its preparing process - Google Patents

Abstract

The present invention belongs to the technical field of a Chinese medicine preparation and discloses fatty emulsion of goldenrod for injection. The emulsion is composed of volatile oil of the effective part of goldenrod, and medical assistant material. Studied by pharmacodynamics, the preparation of the present invention has the functions of analgesia, refrigeration, antiphlogosis and immunity adjustment. The present invention provides a preparing process thereof.

Description

Fatty emulsion of goldenrod for injection and preparation method

Technical field

The invention belongs to the Chinese medicine preparation technical field.Be specifically related to a kind of fatty emulsion of goldenrod for injection and preparation method.

Background technology

Agnogenic heating is one of modal disease of hospital outpatient, emergency treatment, because most and non-bacterial infection, doctor trained in Western medicine does not still have the good curing method at present, often adopts antibiotic therapy now clinically, increased the weight of patient's financial burden, and belonging to antibiotic abuse in fact, its consequence is very serious, even merged bacterial infection, because the cause of disease is comparatively complicated, antibiotic independent application, effect are often dissatisfied, cause state of an illness delay.

A large amount of clinical practices show, these agnogenic fever patients mostly are " going up sense ", medicine among wind and heat dispersing commonly used, the subduing swelling and detoxicating, can obtain good curative effect, but the decoction of Chinese medicine is because of difficulty, unfavorable popularization are stored in transportation, even use electuary, tablets and other formulations, also generate heat higherly because of the patient, and that the Chinese medicine of these dosage forms takes effect is slower, so quite feel inconvenience.

Medicine among several quiet injecting types of part hospital application in recent years, obtained certain effect, but this medicine often has deposited phenomenon, many-sided side effect such as haemolysis, allergy are also arranged clinically, and the medicine that has, so far do not determine the quality standard of its final products as yet, perhaps the aspects such as stability of dosage form remain in certain problem.Along with the development of medicinal career and the progress of Chinese medicine scientific research, the appearance of Chinese medicine of new generation all is badly in need of in society and market.

Herba Solidaginis [Solidago Virgo-aurea L.Var.Leiocarpa (Benth) A.Gray] belongs to feverfew, early be stated from " Zhiwu Mingshi Tukao " and books such as " Compendium of Materia Medica ", the Chinese medicine voluminous dictionary is also included, popular name JINYAOSHI, Huang all over the mountain, HUANGHUACAO etc.The electuary of this medicine is produced and sold as formal product in early days the seventies, and in clinical use.

This property of medicine temperature bitter in the mouth, cure mainly the fiery dispelling wind that looses, heat-clearing and toxic substances removing, plant research has proved that this medicine contains phenol, tannin, volatile oil, Saponin, flavone etc., contain chlorogenic acid (ChlorogenicAcid) in the phenolic constituent, caffeic acid (Caffeic Acid), contain quercetin in the flavonoid, pavilion skin glycoside, globulariacitrin (Kutin), kaempferol glycoside (Radix Astragali glycoside Astragalin), the hot water of two glycosides (Cyainidin3-gentiobioside) herb proposes liquid antibacterial action, its antimicrobial component can be by acid precipitation, and being dissolved in ethanol, it is analgesic that mentioned component also points out this medicine to have, analgesia, antiinflammatory, infection, relieving asthma and immunoregulation effect.

The electuary of using this medicine the seventies has shown that upper respiratory tract infection is had good curative effect, and some hospital once utilized its volatile oil component to make used for intravenous injection upper respiratory tract infection has been shown certain curative effect, have bring down a fever fast, characteristics such as toxicity is little.Stubbornness is infected antibiotic when invalid, add with this medicine disease controlling rapidly.

Summary of the invention

Technical problem to be solved by this invention is that Herba Solidaginis is is further researched and developed, and obtains its active component, expands its clinical practice.

The invention provides fatty emulsion of goldenrod for injection, this lipomul is that active component and pharmaceutic adjuvant are formed with the Herba Solidaginis oil, comprising:

Herba Solidaginis oil 1.0-100g

Injection phospholipid 3-50g

Injection Oleum Glycines 50-300g

Glycerol 25g

Water for injection adds to 1000ml

This product active component is a Herba Solidaginis oil, and concentration is 0.1-10% in the prescription.Adjuvant is an injection Oleum Glycines, and concentration is 5-30% (W/W) in the prescription; Injection phospholipid, concentration is 0.3-5% (W/W) in the prescription; Glycerol and water for injection.Injection Oleum Glycines is as oil phase in prescription, and injection phospholipid is as surfactant, and glycerol is as isotonic agent, and water for injection is as water.

Another technical problem to be solved by this invention is the preparation method of fatty emulsion of goldenrod for injection, and this method comprises the following steps:

1. the Herba Solidaginis oil that takes by weighing recipe quantity is gone in the injection Oleum Glycines of recipe quantity, and mixing is oil phase;

2. the injection phospholipid that takes by weighing recipe quantity is gone in the homogenizer, adds the glycerol of proper amount of water for injection and recipe quantity, starts stirring, and phospholipid is dissolved fully, is water;

3. under high-speed stirred, oil phase is added aqueous phase with wire, treat that oil phase finishes, continue to stir 10 minutes, promptly get milky colostric fluid;

4. colostrum is put in the loading hopper of two step homogenizers, machined, when treating in the pipeline no bubble, first low pressure is adjusted to 5-15kg/cm ²After treating that low pressure is stable, with high voltage adjusting to 30-50kg/cm ², after the circulation for several times Emulsion is transferred in the liquid receiver continuously;

5. the sodium hydroxide solution with 0.1N is adjusted to emulsion about pH7, and reuse water for injection is diluted to 1000ml;

6. emulsion filters with No. 3 sintered filter funnels;

7. carrying out intermediate mass detects;

8. with the emulsion embedding in proper container;

9. the emulsion after the embedding is through 100 ℃ of flowing steam sterilizations 30 minutes, promptly.

The present invention has carried out separation and Extraction to the effective site of Herba Solidaginis, and has done the pharmacodynamics screening, and method is as follows:

(1) volatile oil part

(2) water decoction-alcohol sedimentation n-butanol extraction position

(3) water decoction-alcohol sedimentation macroporous resin ethanol elution position

(4) water decoction-alcohol sedimentation macroporous resin eluent is through silica gel H column chromatography position

Herba Solidaginis extraction process flow process

Herba Solidaginis infection active component intravenous formulations pharmacodynamic study

(1) Herba Solidaginis infection active component drug effect primary dcreening operation

Summary: 4 positions that the Herba Solidaginis extraction process obtains are through analgesia, antiinflammatory, analgesic pharmacological testing, and the effective site that proves its injection for intravenous is volatile oil.

1. experiment purpose:

With 4 effective site pharmacologically actives of analgesia, antiinflammatory, the acquisition of analgesic pharmacology experiment screening Herba Solidaginis extraction process, thus but the component of decision injection for intravenous.

2. be subjected to the reagent thing:

2.1. title: Herba Solidaginis n-butanol portion (B)

Herba Solidaginis macroporous resin column eluting position (E)

Herba Solidaginis volatile oil part (A)

Monomer (salicoside) (F)

2.2. the unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

2.3. content:

Herba Solidaginis n-butanol portion (B) 1g is equivalent to crude drug 27.7g

Herba Solidaginis macroporous resin column eluting position (E) 1g is equivalent to crude drug 17.3g

Herba Solidaginis volatile oil part (A) 1mg is equivalent to crude drug 1.2g

Monomer (salicoside) (F)

2.4. compound method:

Remove Herba Solidaginis volatile oil and be dissolved in the Oleum Arachidis hypogaeae semen China and foreign countries, all the other all are dissolved in 0.5%CMC and are made into suspension.

2.5. dosage setting:

Because of the extractive content difference, dosage is amounted to the crude drug amount and is calculated, and the dose screening of 30g crude drug/kg is got in experiment.

2.6. route of administration: lumbar injection or intravenous injection

2.7. consumption: 0.5ml/20g mice, 1ml/150g rat, 1ml/kg rabbit

2.8. other material:

Proinflammatory agent: carrageenin, 1%, import.

The pyrogenicity agent: the escherichia coli extract, provide for oneself this chamber.

3. animal:

3.1. strain, kind, the quality certification:

Mice, the Kunming kind, male and female all have, 18-20g, Shanghai Institute of Pharmaceutical Industry's Animal House supply, the quality certification: Shanghai is moving closes the card word No. 107.

Rat, SD system, male, 130-150g, Chinese Academy of Sciences's Shanghai Experimental Animal Center supply, the quality certification: No. 005, the moving Guan Huidi of middle section.

Rabbit, male and female all have, body weight 2-2.2kg, Chinese Academy of Sciences's Shanghai Experimental Animal Center supply,

The quality certification: No. 005, the moving Guan Huidi of middle section.

4. contrast medicine:

4.1. aspirin: pharmaceutical factory of Xinhua raw material, this is as positive reference substance.

4.2. Herba Solidaginis aqueous injection: Longhua, Shanghai hospital provides, and 1mI is equivalent to the 2g crude drug, uses for laboratory animal.

4.3. double coptis root injection: Harbin No.2 Traditional Chinese Pharmacetical Plant, lot number 9803172,600mg/ props up, clinical consumption 60mg/kg, i.p gtt.

5. experimental technique:

5.1. the Herba Solidaginis extract active component is to the analgesic activity one acetic acid twisting method of rat

5.1.1. 50 of Kunming mouses, be divided into 5 groups at random, the matched group coordinative solvent, all the other are oral solidago plant style eluent, Herba Solidaginis n-butanol portion, Herba Solidaginis volatile oil and positive control drug aspirin respectively, the equal ip 0.7% acetic acid 10ml/kg of every Mus half an hour after the administration, writhing response appears behind the control group mice ip acetic acid, the distortion body, abdominal part pastes ground, turns round the body number of times in 10 minutes behind the record injection acetic acid after 5 minutes, calculate suppression ratio, compare with matched group.

5.1.2. experimental result:

Experimental result sees Table 1

Writhing response appears behind the control group mice injection acetic acid, the distortion body, and abdominal part pastes ground, the equal obvious suppression of mice of respectively organizing of administration is turned round body action, wherein the mouse writhing of solidago plant style eluent action minimizing is almost nil, and the activity of Herba Solidaginis volatile oil mice is as usual, turns round the body number of times and all reduces.

Each component of table 1 Herba Solidaginis is to the influence of mice acetic acid twisting method

Group	Dosage * approach	Turn round body number of times X ± SD	Suppression ratio %
				Solidago plant style eluent	0.86g/kg ip×1	0**	100
The Herba Solidaginis n-butanol portion	0.54g/kg ip×1	0.6**	96.4
				Herba Solidaginis volatile oil	1.7mg/kg ip×1	5.0±2.74**	83.3
Aspirin	100mg/kg ip×1	4.2±1.6**	86.1
				Matched group	Coordinative solvent	30.1±5.7

Compare with matched group * P＜0.01

5.2. Herba Solidaginis extracts each component of obtaining analgesic activity to mice acetic acid twisting method for the second time

5.2.1. experiment purpose:

Determine again the active of Herba Solidaginis n-butanol portion I, II and monomer, each component of Herba Solidaginis volatile oil and with SHUANGHUANLIAN relatively.

5.2.2 experimental technique:

80 of Kunming mouses, be divided into 8 groups at random, except that matched group gives coordinative solvent, all the other each groups are respectively Herba Solidaginis n-butyl alcohol I, II, monomer, Herba Solidaginis volatile oil, contrast medicine SHUANGHUANLIAN, solidago plant floricome, aspirin, after each treated animal administration half an hour equal lumbar injection 0.7% acetic acid 10mi/kg, turn round the body number of times after 5 minutes in the record hours 10 minutes.

5.2.3 experimental result:

The results are shown in Table 2

What animal each component of Herba Solidaginis behind injection acetic acid all obviously suppressed mice turns round the body number of times, wherein the mice of Herba Solidaginis n-butyl alcohol II is movable obviously reduces, and the mice activity of monomer and Herba Solidaginis volatile oil part is as usual, and animal turns round the body number of times and obviously reduces, and SHUANGHUANLIAN and solidago plant floricome all have certain analgesic activity.

Table 2 Herba Solidaginis component is to the influence of mice acetic acid twisting method

Group	Dosage * approach	Turn round body number of times X ± SD	Suppression ratio %
				Herba Solidaginis n-butyl alcohol I	0.17g/kg ip×1	0.44±0.83**	98.3
Herba Solidaginis n-butyl alcohol II	0.54g/kg ip×1	0**	100
				Monomer	0.05g/kg ip×1	0**	100
Herba Solidaginis volatile oil	1.7mg/kg ip×1	2.33±4.64**	91.48
				SHUANGHUANLIAN	60mg/kg ip×1	6.25±5.1**	77.1
The solidago plant floricome	0.5mg/kg ip×1	11.0±8.8**	59.8
				Aspirin	100mg/kg ip×1	4.10±1.19**	85.02
Matched group	Coordinative solvent	27.37±8.1

Compare with matched group * P＜0.01

5.3. each component of Herba Solidaginis causes acutely inflamed influence to rat by carrageenin

5.3.1. experiment purpose:

Each component of primary dcreening operation Herba Solidaginis has carrageenin to cause acutely inflamed effect to rat

5.3.2 experimental technique:

50 of rats, be divided into 5 groups at random, it is the coordinative solvent group, Herba Solidaginis volatile oil, the Herba Solidaginis n-butanol portion, Herba Solidaginis water extract and aspirin group, survey with drainage earlier before the experiment and respectively organize the right back sufficient normal volume of rat, test and gave the oral medicine feed of each treated animal 1 time the same day, cause inflammation to the right back sufficient sole of the foot intradermal injection 1% carrageenin 0.05ml of each group rat after one hour, mensuration causes scorching back 1,2,3,4,5, right back sufficient volume in hour, the difference that rat causes the right sufficient sole of the foot volume in scorching front and back is the swelling value, calculate swelling value and suppression ratio, and compare with matched group.

E _n=cause the pedal swelling value of scorching back different time

E ₀=cause the pedal swelling value before scorching

The swelling rate of C=matched group

The swelling rate of T=treatment group

5.3.3. experimental result

The results are shown in Table 3, each component of Herba Solidaginis causes the acute inflammation rat to carrageenin all in various degree inhibitory action, but better than volatile oil with the anti-inflammatory activity at Herba Solidaginis n-butanol portion and water extraction position.

Table 3 Herba Solidaginis causes acutely inflamed antiinflammatory action to rat by carrageenin

Group	Cause the swelling value %X ± SD of scorching back different time
						1	2	3	4	5
	Matched group	26.04	47.9	51.35	56.3						45.8
Herba Solidaginis volatile oil 3.4mg/kg ip * 1						21.0 (17.4)	37.0 (22.75)	50.0 (2.6)	41.0 (27.1)	45.6 (0.4)
	Herba Solidaginis n-butanol portion 1.08g/kg po * 1	17.1 (34.4)	33.9 (29.2)	38.2 (25.6)	29.9 (46.8)						31.8 (30.5)
Herba Solidaginis water extract 1.73g/kg po * 1						28.9	27.1 (43.4)	33.2 (35.3)	35.9 (36.2)	27.9 (39.1)
	Aspirin 100mg/kg ip * 1	21.2±3.5 (18.58)	32.3±4.6 (32.5)	30.8±3.9 (40.0)	32.5±4.1 (42.2)						31.1±4.5 (32.1)

In () is suppression ratio

5.4. each component of Herba Solidaginis is to the influence of rabbit by escherichia coli pyrogenicity

5.4.1. experiment purpose:

Each component of primary dcreening operation Herba Solidaginis is to the refrigeration function of rabbit by escherichia coli pyrogenicity.

5.4.2. experimental technique:

Get to survey body temperature and to be stabilized in 38.5-39.6 ℃ rabbit, coli somatic by the intravenous injection deactivation, 0.5-1hr 18 of the fever in rabbit of back selective body temperature rise more than 1 ℃, be divided into 6 groups at random, difference intravenous injection Herba Solidaginis volatile oil, Herba Solidaginis water distillate, monomer, Herba Solidaginis n-butanol portion and aspirin, matched group gives normal saline, surveys body temperature 1 time by anus in per 1 hour after the administration.

5.4.3. experimental result:

The results are shown in Table 4, rabbit body temperature behind the injection coli somatic rises rapidly, matched group body temperature rose very fast at 1-2 hour, decline is arranged after 3 hours slightly, and Herba Solidaginis water distillate and monomer body temperature rise and decline are more even, dead immediately after the Herba Solidaginis n-butanol portion rabbit ip injection, have only the Herba Solidaginis volatile oil part to inject back 2 hours body temperature and begin to descend in the pyrogenicity agent, aspirin group body temperature after 2 hours begins to descend.

Each component of table 4 Herba Solidaginis is to the influence of rabbit escherichia coli pyrogenicity

Group	“0” ℃	The pyrogenicity agent is injected back 0 ℃	The body temperature of different time after the administration
								1	2	3	4	5
			Matched group	39.3	40.7	41.4	41.6						40.9	40.5	40.5
Herba Solidaginis volatile oil 1.93mg/kg	39.3	40.3						40.0	39.7	39.8	39.5	39.4
			Herba Solidaginis water distillate	39.2	40.2	40.3	40.2						40.2	40.3	40.2

1ml/kg
								Monomer 25mg/kg	39.3	40.3	40.5	40.4	40.5	41.2	40.7
Herba Solidaginis n-butanol portion 1g/kg	39.3	40.3	Dead immediately
								Aspirin 100mg/kg	39.1	40.2	40.1	40.0	39.9	39.5	39.2

(2) each component hemolytic test of Herba Solidaginis

1. experiment purpose:

Observe the haemolysis situation of each component of Herba Solidaginis, thereby decision can be for the effective site of intravenous formulations to rabbit.

2. animal: rabbit, body weight 2.5kg, Chinese Academy of Sciences's Shanghai Experimental Animal Center supply.

The quality certification: No. 005, the moving Guan Huidi of middle section.

3. given the test agent:

Herba Solidaginis n-butanol portion: concentration 10mg/ml

Solidago plant style eluate: concentration 10mg/ml

Herba Solidaginis volatile oil: concentration 10mg/ml

4. solvent: normal saline

5. experimental technique:

Get fresh Sanguis Leporis seu oryctolagi 2ml, stir with the removal fibrin with bamboo let, reuse normal saline flushing 3-5 time, centrifugal, no longer present until the supernatant redness, the centrifuging and taking precipitation is pressed the gained red cell volume, is made into 2% suspension with normal saline.Add various solution with following method, the 6th pipe does not add test sample, and as blank, the 7th pipe adding distil water contrasts as haemolysis, 1 hour observed result of insulation in 37 ℃ of water-baths.

Test tube	1	2	3	4	5	6	7
								Need testing solution (ml)	0.1	0.2	0.3	0.4	0.5	1
Normal saline (ml)	2.4	2.3	2.2	2.1	2.0	2.5	Distilled water 2.5
								2% red cell suspension (ml)	2.5	2.5	2.5	2.5	2.5	2.5	2.5

6. result:

Herba Solidaginis n-butanol portion: haemolysis

Solidago plant style eluate: haemolysis

Herba Solidaginis volatile oil: haemolysis not

7. conclusion:

Because Herba Solidaginis n-butanol portion and the eluate of solidago plant style all have haemolysis to produce, and produce dead to rabbit vein injection back.Further extraction can't separate the haemolysis component again, thus only take Herba Solidaginis volatile oil as the effective active component, the preparation of preparation injection for intravenous.

The research of Herba Solidaginis volatile oil component and quality control standard

Volatile oil is the very complicated mixture of some compositions normally, but requirement according to two class Chinese medicines, its composition is carried out suitable research be absolutely necessary, we adopt the method for gas chromatogram and gas chromatogram-mass spectrometry that the composition of Herba Solidaginis volatile oil is studied for this reason.

(1) physicochemical property of Herba Solidaginis volatile oil

Herba Solidaginis volatile oil color is light brown, and the physicochemical property that secondary is measured is as follows:

Proportion optically-active index of refraction

0.96 [α] ₂₀-11.16°o 1.5036

0.95 [α] ₂₀-10.34°o 1.5015

(2) Herba Solidaginis volatile oil component research

Volatile oil is for containing multiple mixture of ingredients, and it forms extremely complicated, and tens kinds of chemical compounds to hundreds of are arranged usually.For the analysis of volatile oil component, gas chromatography is compared with the method for measuring physico-chemical constant, has strong, the highly sensitive characteristics of separation efficiency.For this reason, adopt the temperature programming capillary gas chromatography, to Herba Solidaginis volatile oil respectively at non-polar column (HP-1, HP-5) and on the polar column (SUPELCOWAX) carried out analysis and investigation, thereby set up the gas chromatogram method for separating and analyzing, each composition in the Herba Solidaginis volatile oil is separated, and adopt area normalization method to determine the percentage composition of each composition.

Qualitative investigation work about each component: the qualitative analysis of volatile oil is generally adopted standard control, list of references value or is carried out in conjunction with other analysis means.Owing to except that indivedual compositions, be difficult to obtain the standard substance of composition, and literature value is not only few but also loose.Therefore adopted the method for gas chromatography and mass spectromentry coupling, the high score by gas chromatogram in conjunction with the mass spectrum library searching, carries out qualitative investigation to the Herba Solidaginis volatile oil component from the function of ability and mass spectrometry method authenticating compound.Below be that present completed work is concluded.

1. the gas chromatographic analysis of Herba Solidaginis volatile oil

The selection of chromatographic condition:

In order to realize multi-component good separation, adopt the temperature programming capillary gas chromatography, select the different chromatographic column of polarity to investigate volatile oil (sample number into spectrum 1 ^#) in the chromatographic behavior of composition, (HP-1, HP-5) separating effect is relatively poor, and goes up good separating effect in polarity chromatographic column (SUPELCOWAX) in nonpolar chromatographic column for sample.Parameters such as reprogramming initial temperature, carrier gas linear speed, heating rate are observed the suitability of chromatographic condition respectively, have determined the gas chromatogram compartment analysis condition of Herba Solidaginis volatile oil.

2. the qualitative investigation of Herba Solidaginis volatile oil

Herba Solidaginis volatile oil separates its composition through capillary gas chromatography, but is difficult to obtain standard substance owing to remove indivedual compositions (β-caryophyllene), by the chromatography retention time each component is contrasted discriminating.In view of gas chromatogram-mass spectrometric hyphenated technique is a prefered method of analyzing the Chinese herbal medicine volatile oil component now, we adopt the gas chromatography mass spectrometry technology that the composition of Herba Solidaginis volatile oil has been carried out qualitative investigation for this reason.

(1) instrument and experiment condition:

Instrument: FinniganMATGCQ gas chromatograph-mass spectrometer

Chromatographic condition: chromatographic column: Supelcowax 30m 0.25mmlD o.25film

Carrier gas: He split ratio: 80: 1 injector temperatures: 230 ℃

Temperature programming condition: 50 ℃ of (hold3min) → 2 ℃/min → 160 ℃

(hold9min)→3℃/min→200℃

The mass spectrum condition:

Ionization mode: El. ion source temperature: 190 ℃

Electron energy: 70eP. quality of scanning scope: 25-350

Resolution: 1000

The GCQ data handling system

(2) experimental result:

Herba Solidaginis volatile oil 1 ^#Sample is analyzed through GC-MS, obtain directly to carry out library searching after the mass spectrometric data by the instrument data processing system, further check again, thereby differentiated the Herba Solidaginis main component of essential oil with mass spectrum standard diagram EPA/NIHMass Spetral Data Base and The Wiley/NBS Registry Of MassSpectral Data.

Gained total ion current figure and experimental result are as shown in table 4:

Table 4 Herba Solidaginis chemical composition of volatile oil and content

The peak numbering	M +	Chemical compound
				1	136	3-Carene	The 3-carene
2	136	Sabinene	Sabinene (mircene)
				3	136	Limonene	Limonene
4	204	α-Cadinene	Cadinene
				5	204	α-Copaene
6	204	β-Cubebene
				7	204	β-Caryophyliene	The p-caryophyllene
8	204	α-Guaiene
				9	204	Germacrene D	Big Mang bicyclogermacrene
10	204	δ-Cadinene
				11	202	Calamnene<1s，cis->
12	200	α-Calacorene or β-Calacorene
				13	220	Caryophyliene cxide	The oxidation caryophyllene
14	220	Thujopsanone

15	220	Unknown
				16	220	Spathulenol	Spathulenol
17	220	6，9-Octadecadiyoic acid，methyl ester
				18	220	1H-Cycloprop[e]azulen-7-ol， decahydro-1，1，7-trimethyl-4-methlen	Encircle third Austria-7-alcohol
19	220	Unknown
				20	220	cis-.alpha.-Copaene-8-ol
21	220	Cedran-13-ol<8->acetate
				22	220	Unknown
23	220	Unknown

(4) Various Seasonal, lot number sample pharmacodynamic study

The content of considering the above-mentioned several compositions of different lot numbers (being Various Seasonal institute sample thief) is not.Unanimity is so decision is with 7 ^#With 8 ^#Sample carries out pharmacodynamics and measures.

The result is as follows:

The analgesic effect of table 5 20010820 Dichlorodiphenyl Acetate writhing methods

Group	Dosage * approach	Turn round body number of times X ± SD	Suppression ratio %
				Herba Solidaginis 20010820	1mg/kg ip×1	16.4±1.2	15.0
2mg/kg ip×1	14.2±1.8	26.4
			4mg/kg ip×1		11.3±4.6 *	41.4
Aspirin	100mg/kg ip×1	4.9±2.1 **					74.6
			Matched group	Coordinative solvent	19.3±4.3

^*Compare with matched group P＜0.01.

20010820 lot number curative effects can not show a candle to former experimental result.

The analgesic effect of the different lot number Dichlorodiphenyl Acetate of table 6 Herba Solidaginis volatile oil writhing method

Group	Dosage mg/kg	Turn round body number of times X ± SD	Suppression ratio %
				Mix and criticize	4	1.21±1.08	95.4
7 # Sample	4	0.47±0.36	98.2
				8 # Sample	4	1.05±0.89	96.0
Matched group		26.30±7.05

Mix and criticize: 7 ^#With 8 ^#The mixing of sample

The analgesic effect of table 7 20011030 Dichlorodiphenyl Acetate writhing methods

Group	Dosage * approach	Turn round body number of times X ± SD	Suppression ratio %
				Herba Solidaginis	1mg/kg ip×1	9.0±3.5 **	70

20011030	2mg/kg ip×1	4.5±2.3 **	85
				4mg/kg ip×1	0.6±0.2 **	98
	Aspirin	100mg/kg ip×1	6.4±3.2 **				79
Matched group				Coordinative solvent	30.1±5.6

^*Relatively annotate with matched group P＜0.01: 20011030 promptly mix batch.

3. brief summary:

Through the work of having carried out, what time followingly sum up:

(1) adopt the temperature programming capillary gas chromatography preferably compartment analysis the composition of Herba Solidaginis volatile oil, contained component is many, and be that some boiling points are very approaching mostly, or the chemical compound of isomers, adopt the peak area normalization method that Herba Solidaginis volatile oil is carried out quantitative analysis at present, what obtain is the relative percentage composition of each component.

(2) gas chromatography and mass spectromentry coupling method is one of effective method of the unknown chemical constituent of isolation identification.Identified Herba Solidaginis volatile oil main component by gas one matter multiple techniques and library searching.

(3) experiment shows, the Herba Solidaginis volatile oil that extracts from the same places of origin of raw materials and collection season has similar component, and the volatile oil component of Various Seasonal (lot number) is basic identical, and is just variant on the components contents.The higher several components of content are p-caryophyllene (about 75 minutes of about 36.7 minutes of retention time of β-Caryophyliene), about 42.5 minutes of big Mang bicyclogermacrene retention time, about 59.3 minutes of oxidation caryophyllene (Caryophyliene oxide) retention time, about 66 minutes of spathulenol retention time, ring third Austria-7-alcohol retention time through differentiating.Through the mensuration of several batch samples is found that these several components contents just have nothing in common with each other, but their content sums are between 34-43%.

The preliminary pharmacodynamic study of Herba Solidaginis

1. the analgesic activity of Herba Solidaginis

1.1. experiment purpose:

Confirm the analgesic effect of Herba Solidaginis preparation.

1.2. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, what obtain is water-soluble

The property injection, every injection 1ml contains volatile oil 4mg, but injection for intravenous is used.

1,2,4mg/kg dosage:

Consumption: 0.5ml/20g mice

Usage: iv * 1

1.3. animal:

Source, strain, the quality certification: mice, the Kunming kind, male, 19-21g, Shanghai Institute of Pharmaceutical Industry's Animal House supply.

The quality certification: Shanghai is moving closes the card word No. 107.

1.4. contrast medicine: aspirin, this chamber are provided (Xinhua's pharmaceutical factory raw material) for oneself

SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, lot number 980372, every contains

600mg, dosage 60mg/kg, iv, gtt.

1.5. experimental technique:

60 of Kunming mouses, be divided into 6 groups, Herba Solidaginis is respectively 1,2,4mg/kg, SHUANGHUANLIAN 60mg/kg, aspirin 100mg/kg, matched group gives coordinative solvent, be iv * 1, after the administration half an hour every Mus ip 0.3% acetic acid 10ml/kg, the mouse writhing number of times of record in 10 minutes after 5 minutes, and with matched group relatively.

1.6. experimental result:

The results are shown in Table 1, Herba Solidaginis has obvious suppression acetic acid to cause the pain of mice, and has---and fixed dosage effect, SHUANGHUANLIAN and aspirin also all have the effect of obvious suppression pain.

Table 8 Herba Solidaginis is to the analgesic activity of mice method of acetic acid

Group	Dosage * approach	Turn round body number of times X ± SD	Suppression ratio %
				Herba Solidaginis	1mg/kg ip×1	16.9±5.5 **	44.5
Herba Solidaginis	2mg/kg ip×1	10.4±3.1 **	65.9
				Herba Solidaginis	4mg/kg ip×1	6.50±2.5 **	78.6
SHUANGHUANLIAN	60mg/kg ip×1	11.7±6.64 **	61.6
				Aspirin	100mg/kg ip×1	6.4±3.2 **	79.0
Matched group	Coordinative solvent	30.5±5.78

^*Compare with matched group P＜0.01

2. Herba Solidaginis is influenced by the exothermic reaction that escherichia coli cause rabbit

2.1. experiment purpose:

Observe the refrigeration function of Herba Solidaginis, and make comparisons with SHUANGHUANLIAN.

2.2. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1ml contains volatile oil 4mg, but injection for intravenous is used.

Dosage: 2,4mg/kg

Consumption: 0.5ml/kg

Usage: iv * 1

2.3. contrast medicine: aspirin, this chamber are provided (Xinhua's pharmaceutical factory raw material) for oneself

SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

2.4. pyrogenicity agent: coli somatic, this chamber prepares voluntarily.

2.5. animal: source, strain, the quality certification:

Rabbit, male and female all have, 2.0-2.2kg, Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.

The quality certification: No. 005, the moving Guan Huidi of middle section.

2.6. experimental technique:

Get to survey body temperature and to be stabilized in 38.5-39.6 ℃ rabbit, coli somatic by the intravenous injection deactivation, 0.5-1hr 15 of the fever in rabbit of back selective body temperature rise more than 1 ℃, be divided into 5 groups at random, Herba Solidaginis intravenous injection 2 respectively, 4mg/kg, SHUANGHUANLIAN 60mg/kg, aspirin 100mg/kg, matched group gives normal saline, surveys body temperature 1 time by anus in per 1 hour after the administration, observes body temperature and changes.

2.7. experimental result:

The results are shown in Table 9, body temperature rises rapidly after the agent of matched group rabbit injection pyrogenicity, and is basicly stable in after 6 hours, gives in Herba Solidaginis treated animal 1-3 hour body temperature rise slower, and beginning in 4 hours descends gradually, and the decline of SHUANGHUANLIAN treated animal body temperature is not obvious.

Table 9 Herba Solidaginis is to the influence of rabbit by escherichia coli pyrogenicity

Group	Normal body temperature ℃	The pyrogenicity agent is injected back 0 ℃	The body temperature of different time after the administration
									1	2	3	4	5	6
			Matched group	39.1	40.1	40.4	40.4	40.3							40.5	40.5	40.5
SHUANGHUANLIAN	39.3	40.3							40.4	40.4	40.4	39.5	39.4	40.1
			Herba Solidaginis 4mg/kg	38.9	39.9	40.0	39.9	39.8							40.3	40.2	39.4
Herba Solidaginis 2mg/kg	38.8	39.8							39.8	39.9	39.8	41.2	40.7	39.4
			Aspirin 100mg/kg	39.1	40.1	40.0	39.9	39.5							39.5	39.2	39.2

3. Herba Solidaginis causes acutely inflamed inhibitory action to rat by carrageenin

3.1. experiment purpose:

Observe the Herba Solidaginis injection rat is caused acutely inflamed antiinflammatory action.

3.2. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1mi contains volatile oil 4mg, but injection for intravenous is used.

1,2,4mg/kg dosage:

Consumption: a 1ml/ rat

Usage: ip * 1

3.3. animal: source, strain, the quality certification:

Rat, SD system, 130-150g, male, Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.

The quality certification: No. 005, the moving Guan Huidi of middle section.

3.4. contrast medicine: aspirin, this chamber are provided (Xinhua's pharmaceutical factory raw material) for oneself

SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

3.5. proinflammatory agent: carrageenin, 1%, import.

3.6. experimental technique:

50 of rats, be divided into 5 groups at random, be that coordinative solvent group, Herba Solidaginis are respectively 1mg/kg, 2mg/kg, 4mg/kg and aspirin 100mg/kg, survey with drainage earlier before the experiment and respectively organize the right back sufficient normal volume of rat, test and gave the oral medicine feed of each treated animal 1 time the same day, cause inflammation to the right back sufficient sole of the foot intradermal injection 1% carrageenin 0.05ml of each group rat after one hour, mensuration cause scorching back 1,2,3,4,5, hour in right back sufficient volume, the difference that rat causes the right sufficient sole of the foot volume in scorching front and back is the swelling value, calculate swelling value and suppression ratio, and compare with matched group.

E _n=cause the pedal swelling value of scorching back different time

E ₀=cause the pedal swelling value before scorching

The swelling rate of C=matched group

The swelling rate of T=treatment group

3.7. experimental result:

The results are shown in Table 3, Herba Solidaginis causes scorching rat to carrageenin in various degree inhibitory action, and presents certain dose-effect relationship, and positive drug aspirin also has the effect of inflammation-inhibiting.

Table 10 Herba Solidaginis causes acutely inflamed antiinflammatory action to rat by carrageenin

Group	Cause the swelling value %X ± SD of scorching back different time
							1	2	3	4	5
Matched group	24.5×0.9	35.8×1.0	41.5×1.57	34.9×2.4	37.7×2.1
						Herba Solidaginis 1mg/kg ip * 1	22.6×3.1 (7.75)	29.9×2.9 (16.48)	35.3×2.75 (14.93)	28.8×3.0 * (17.47)	36.03×2.4 (4.42)

Herba Solidaginis 2mg/kg ip * 1	21.2×2.8 (13.46)	29.3×1.47 ** (18.15)	29.1×1.02 ** (28.87)	24.7×1.25 ** (29.22)	25.5×1.34 ** (32.55)
						Herba Solidaginis 4mg/kg ip * 1	19.5×1.5 (20.4)	25.4×1.70 * (29.05)	23.8×2.3 ** (42.65)	22.8×2.1 ** (34.67)	21.9×1.65 ** (41.88)
Aspirin 100mg/kg ip * 1	18.2×3.5 (25.71)	24.4×4.6 ** (31.84)	23.8×2.9 ** (31.84)	21.4×3.1 ** (38.68)	21.8×2.8 ** (42.17)

P＜0.05, ^*Compare with matched group P＜0.01

4. Herba Solidaginis is to effect of immunologic function

4.1. the influence that the Herba Solidaginis injection transforms mouse spleen lymphocyte (in the body)

Summary: the Herba Solidaginis injection can obviously promote mouse spleen lymphocyte is transformed, and certain dose-effect relationship is arranged.

4.1.1. experiment purpose:

The influence that test Herba Solidaginis injection transforms mouse spleen lymphocyte.

4.1.2. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1mI contains volatile oil 4mg, but injection for intravenous is used.

Dosage: 2,4mg/kg

Consumption: 0.5ml/20g mice

Usage: ip * 6

4.1.3. animal: source, strain, the quality certification:

Mice, 057BEl6,19-21g, male, 5 every group.Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.The quality certification: No. 005, the moving Guan Huidi of middle section.

4.1.4. contrast medicine: SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

4.1.5. other material:

Canavaline ConA:sigma, 50 μ g/ml.

³H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml.

1640 culture medium: Difco product

4.1.6. experimental technique:

C ₅₇The BL/6 mice, random packet, Herba Solidaginis gives 2mg/kg, 4mg/kg, SHUANGHUANLIAN 60mg/kg respectively, matched group is a normal saline, is lumbar injection, every day 1 time, totally 6 times, after administration finishes, put to death animal, get spleen under the aseptic condition, the counting splenocyte, and the adjustment cell concentration is 1 * 10 ⁷/ ml, every hole adds cell suspension 100 μ l on 96 porocyte culture plates, and ConA 50 μ l and culture fluid are all established three multiple holes, 37 ℃, 5%CO for every group ₂Cultivated 48 hours under the condition, add ³H-TdR 0.5 μ ci/ hole continues to cultivate 18 hours, with bull cell harvestor collecting cell, surveys the CPM value on liquid scintillation instrument, and compares with matched group.

4.1.7. result:

The results are shown in Table 11.

The Herba Solidaginis injection transforms 2, has than obvious facilitation during 4mg/kg mouse spleen lymphocyte.

The influence that table 11 Herba Solidaginis transforms mouse spleen lymphocyte (in the body)

Group	Dosage * approach	The CPM value (X ± SD)
			Herba Solidaginis	2mg/kg ip×6	9775±5177 **
Herba Solidaginis	4mg/kg ip×6	26843±1464 **
			SHUANGHUANLIAN	60mg/kg ip×6	5453±1486 **
Matched group	Coordinative solvent	1267±74

^*Compare with matched group P＜0.01

4.2. the influence (external) that Herba Solidaginis transforms mouse spleen lymphocyte

4.2.1. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1 ' 8 mixed after, the water solublity injection that obtains, every injection 1mI contains volatile oil 4mg, but injection for intravenous is used.

4.2.2. animal: source, strain, the quality certification:

Mice, C ₅₇BL/6,19-21g, male, Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.

The quality certification: No. 005, the moving Guan Huidi of middle section.

4.2.3. contrast medicine: SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

4.2.4. other material:

Canavaline ConA:sigma, 50 μ g/ml.

³H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml.

1640 culture medium: Difco product

4.2.5. experimental technique:

C ₅₇Get spleen under the BL/6 mice, aseptic condition, separating Morr. cell, and the adjustment cell concentration is 1 * 10 ⁷/ ml, Herba Solidaginis becomes 1,0.1,0.01,0.001 μ g/ml with the culture medium mice, and SHUANGHUANLIAN is diluted to 6,0.6,0.06,0.006 μ g/ml, every hole adds various medicinal liquid 100 μ l, ConA 50 μ l, culture medium 100 μ l on 96 porocyte culture plates, 37 ℃, 5%CO ₂Cultivated 48 hours under the condition, add ³H-TdR 0.5 μ ci/ hole continues to cultivate 18 hours, with bull cell harvestor collecting cell, surveys the CPM value on the liquid scintillation instrument, and compares with matched group.

4.2.6. result:

The results are shown in Table 12, Herba Solidaginis transforms mouse spleen lymphocyte in various dose different effects, and SHUANGHUANLIAN also has same purpose.

The influence (external) that table 12 Herba Solidaginis transforms mouse spleen lymphocyte

Group	Concentration	The CPM value (X ± SD)
			Herba Solidaginis	0.001μl/ml	21354±544 **
0.01μl/ml	16171±2855 **
		0.1μl/ml		195±63
1μl/ml	166±26
		SHUANGHUANLIAN		0.006μg/ml	15661±1498 **
0.06μg/ml	194834±740 **
			0.6μg/ml	388±232
6μg/ml	301±139
			Matched group	N.S.	1470±45

^*Compare with matched group P＜0.01

4.3. Herba Solidaginis is to the influence of mice natural killer cell (NK cell)

Summary: Herba Solidaginis has certain activation to NK cells in mice.

4.3.1. experiment purpose:

Test Herba Solidaginis injection is to the influence of NK cells in mice.

4.3.2. supply test agent:

Title: Herba Solidaginis

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1ml contains volatile oil 4mg, but injection for intravenous is used.

Dosage: 2,4mg/kg

Consumption: 0.5ml/20g mice

Usage: ip * 6

4.3.3. animal: source, strain, the quality certification:

Mice, C ₅₇BL/6,19-21g, female, 5 every group.

Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.

The quality certification: No. 005, the moving Guan Huidi of middle section.

4.3.4. contrast medicine: SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

4.3.5 other material:

The YAC-1 cell: pharmacological room of Shanghai Institute of Pharmaceutical Industry goes down to posterity.

1640 culture medium: Difco product

³H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml.

4.3.6. experimental technique:

The C57BL/6 mice, random packet, Herba Solidaginis is respectively 2mg/kg, 4mg/kg, SHUANGHUANLIAN 60mg/kg, and all animal after the last administration finishes, is put to death in ip * 6, gets spleen under the aseptic condition, the counting splenocyte, and the adjustment cell concentration is 1 * 10 ⁶/ ml action effect cell, other gets the YAC-1 cell of cultivating 24 hours, and adjusting cell concentration is 1 * 10 ⁴/ ml is as target cell, is that 1: 100 concentration is added on the 96 porocyte culture plates with target cell and effector lymphocyte's ratio, adds ³Three multiple holes, 37 ℃ of 5%CO are all established for every group in H-TdR 0.5 μ ci/ hole ₂Cultivated 24 hours under the condition, collecting cell is surveyed the CPM value, calculates specificity percentage rate (pi) expression NK cytoactive.

4.3.7. result:

The results are shown in Table 13, Herba Solidaginis has certain facilitation to NK cells in mice, and SHUANGHUANLIAN also has certain effect.

Table 13 Herba Solidaginis is to the influence (in the body) of NK cells in mice

Group	Dosage * approach	The CPM value (X ± SD)	The active % of NK
				Herba Solidaginis	2mg/kg ip×6	11680±820 **	19.8
Herba Solidaginis	4mg/kg ip×6	9520±970 **	34.6
				SHUANGHUANLIAN	60mg/kg ip×6	10768±615 **	26.1
Matched group	Coordinative solvent	14572±780

^*Compare with matched group P＜0.01

4.4. Herba Solidaginis is to the influence (in vitro method) of NK cells in mice

4.4.1. supply test agent:

Title: Herba Solidaginis injection

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+Tween 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1mI contains volatile oil 4mg, but injection for intravenous is used.

4.4.2. animal: source, strain, the quality certification:

Mice, C ₅₇BL/6,19-21g, male, Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.The quality certification: No. 005, the moving Guan Huidi of middle section.

4.4.3. contrast medicine: SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

4.4.4. other material:

The YAC-1 cell: pharmacological room of Shanghai Institute of Pharmaceutical Industry goes down to posterity.

³H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml.

1640 culture medium: Difco product

4.4.5. experimental technique:

C ₅₇The BL/6 mice, get spleen under the aseptic condition, adjusting cell concentration is 1 * 106/ml action effect cell, other gets specimen and adjusts that concentration is 1,0.1,0.01,0.001gg/ml, SHUANGHUANLIAN adjusts that concentration is 6,0.6,0.06,0.006gg/mI, and other gets the YAC-1 cell of cultivating 24 hours and adjusts cell concentration is 1 * 10 ¹/ ml as target cell, is that 1: 100 ratio adds on the 96 porocyte culture plates with target cell and effector lymphocyte's ratio, adds ³Three multiple holes, 37 ℃, 5%CO are all established for every group in H-TdR0.5 μ ci/ hole ₂Cultivated 24 hours under the condition, collecting cell is surveyed the CPM value, calculates specificity percentage rate (pi) expression NK cytoactive.

4.4.6. result:

The results are shown in Table 14, Herba Solidaginis presents different effects to NK cells in mice at variable concentrations.

Table 14 Herba Solidaginis is to the influence (external) of NK cells in mice

Group	Concentration	The CPM value (X ± SD)	The active % of NK
				Herba Solidaginis	0.001μl/ml	3298±398 **
0.01μl/ml	5596±96 **
			0.1μl/ml		1010±119	72.5
1μl/ml	109±37	97.0
			SHUANGHUANLIAN		0.006μg/ml	3717±290 **
0.06μg/ml	4197±256 **
				0.6μg/ml	1486±89	59.5
6μg/ml	127±11	96.5
				Matched group	N.S.	3678±278

^*Compare with matched group P＜0.01

4.5. the influence that Herba Solidaginis produces mice IL-2

Summary: Herba Solidaginis presents inhibitory action when the low dosage to mice IL-2.

4.5.1 experiment purpose:

Observe Herba Solidaginis produces IL-2 to mice influence.

4.5.2 supply test agent:

Title: Herba Solidaginis

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Composition: Herba Solidaginis+temperature 80 with 1: 8 mixed after, the water solublity injection that obtains, every injection 1ml contains volatile oil 4mg, but injection for intravenous is used.

Dosage: 2,4mg/kg

Consumption: 0.5ml/20g mice

Usage: ip * 6

4.5.3 animal: source, strain, the quality certification:

Mice, C ₅₇BL/6,19-21g, female, Chinese Academy of Sciences's Shanghai Experimental Animal Center provides.

The quality certification: No. 005, the moving Guan Huidi of middle section.

4.5.4. contrast medicine: SHUANGHUANLIAN, Harbin No.2 Traditional Chinese Pharmacetical Plant, 600mg/ props up, 60mg/kg,

Lot number 980372.

4.5.5. other material:

Ctll cell: pharmacological room of Shanghai Institute of Pharmaceutical Industry provides for oneself.

Culture medium: 1640, the Difco product

³H-TdR: Shanghai nuclear research institute, radioactive concentration 1mci/ml.

4.5.6. experimental technique:

C ₅₇20 of BL/6 mices are divided into 4 groups at random, and Herba Solidaginis is respectively 2mg/kg, 4mg/kg, SHUANGHUANLIAN 60mg/kg and matched group, control group administered physiological saline, animal after administration finishes, is put to death in the every group of equal ip of medicine * 6, get spleen under the aseptic condition, separating Morr. cell, and to adjust cell concentration be 1 * 107/mi, every hole adds cell suspension 2ml and ConA 50 μ g/ml (final concentration) on 24 porocyte culture plates, 37 ℃, 5%CO ₂Cultivated under the condition 24 hours, and collected supernatant as testing sample, with IL-2 dependent cells CTLL with ³H-TdR mixes method and surveys CPM value, the CPM value of comparative control group and administration group.

4.5.7 experimental result:

The results are shown in Table 15, Herba Solidaginis produces IL-2 to mice and produces inhibitory action.

Table 15 Herba Solidaginis produces the influence (in the body) of IL-2 to mice

Group	Dosage * approach	The CPM value (X ± SD)
			Herba Solidaginis	2mg/kg ip×6	3890±311 **
Herba Solidaginis	4mg/kg ip×6	5090±149 *
			SHUANGHUANLIAN	60mg/kg ip×6	3776±596 **
Matched group	Coordinative solvent	6150±143

^*P＜0.05, ^*Compare with matched group P＜0.01

The research of Herba Solidaginis antivirus action

Herba Solidaginis external discovery under 250pg/ml concentration has the obvious suppression effect to influenza virus and adenovirus, does SHUANGHUANLIAN simultaneously and compares (1mg/ml concentration) and also do not see that SHUANGHUANLIAN has tangible effect.

The research of Herba Solidaginis antifungic action

External under 200 μ g/ml concentration with Herba Solidaginis, to Candida albicans, neogenesis cryptococcus, trichophyton, the equal unrestraint effect of Gypsum Fibrosum trichophyton, make SHUANGHUANLIAN simultaneously under 1mg/ml concentration, also do not see the obvious suppression effect.

Herba Solidaginis is to the inhibitory action of antibacterial

Herba Solidaginis has bacteriostasis to Diplococcus pneumoniae under 30 μ g/ml concentration, SHUANGHUANLIAN also has bacteriostasis being low to moderate under the 30 μ g/ml concentration.

Herba Solidaginis does not all have the obvious suppression effect to staphylococcus aureus, escherichia coli etc.

The research of Herba Solidaginis acute toxicity

With 20 mices of Herba Solidaginis Intravenous injection solution 0.5ml (containing 2.5mg volatile oil) intravenous injection (body weight 18-20g, each 10 of male and female), observe a week, no animal dead, not seeing has any abnormal response, so the LD of Herba Solidaginis ₅₀Greater than 125mg/kg.

Herba Solidaginis lipomul pharmacodynamics test

Summary: the mice analgesic test proved that it has significant analgesia role after Herba Solidaginis was prepared into lipomul.

1. experiment purpose:

The Herba Solidaginis lipomul is to the analgesic activity of mice acetic acid twisting method.

2. testing sample:

Title: Herba Solidaginis lipomul

The unit of providing: Shanghai Institute of Pharmaceutical Industry's mesothecium

Content: every 1mi contains the 8mg Herba Solidaginis

3. animal:

Source, strain, the quality certification:

Mice, the Kunming kind, 19-21g, male, Shanghai Institute of Pharmaceutical Industry's Animal House provides, the quality certification: Shanghai is moving closes the card word No. 107.

4. experimental technique:

40 of Kunming mouses, be divided into 4 groups at random, every group 10, be divided into Herba Solidaginis lipomul 2mg/kg, 4mg/kg, be iv * 1, aspirin 100mg/kg, ip * 1, matched group gives normal saline, after the administration half an hour ip 0.7% acetic acid 10ml/kg, that surveyed mice in 10 minutes in 5 minutes turns round the body number of times.

5. experimental result:

The results are shown in Table 16, mice iv still can obviously suppress the pain reaction that mice causes chemical irritant one acetic acid after giving lipomul, and result and Herba Solidaginis aqueous injection result are similar.

Table 16 Herba Solidaginis lipomul is to the influence of mice acetic acid twisting method

Group	Dosage * approach	Turn round body number of times (X ± SD)	Suppression ratio %
				The Herba Solidaginis lipomul	2mg/kg ip×6	9.0±5.45	57.7
The Herba Solidaginis lipomul	4mg/kg ip×6	4.5±5.9	78.9
				Aspirin	100mg/kg ip×6	4.2±3.2	80.3

Matched group

Coordinative solvent

21.3±5.6

The above results shows that fatty emulsion of goldenrod for injection has effects such as analgesic, antiinflammatory of analgesia and adjusting immunologic function, and the medicine valency is more honest and cleaner, and bigger economic benefit and social benefit will be arranged.

Description of drawings

Fig. 1 granule number scattergram (1)

TSM ultra-fine grain Particle Size Analyzer, Shanghai University of Science and Technology
			Shanghai Institute of Pharmaceutical Industry's sample name: YZHH date: 06-03-2002 time: 14:56:40 sequence number: 1 D32=0.5376 Dv05=0.554 Dv01=0.459 Dv09=0.624 specific surface area: 11.161
Size (μ m) distribution % accumulation % 0.05-0.17 0.00 0.00 0.17-0.29 0.10 0.10 0.29-0.41 3.04 3.14 0.41-0.53 30.79 33.92 0.53-0.65 61.94 95.86 0.65-0.76 4.14 100.00 0.76-0.88 0.00 100.00 0.88-1.00 0.00 100.00 1.00-1.12 0.00 100.00 1.12-1.24 0.00 100.00 1.24-1.36 0.00 100.00 0.36-1.48 0.00 100.00 1.48-1.60 0.00 100.00 1.60-1.72 0.00 100.00 1.72-1.84 0.00 100.00 1.84-1.95 0.00 100.00 1.95-2.07 0.00 100.00	Size (μ m) distribution % accumulation % 2.07-2.19 0.00 100.00 2.19-2.31 0.00 100.00 2.31-2.43 0.00 100.00 2.43-2.55 0.00 100.00 2.55-2.67 0.00 100.00 2.67-2.79 0.00 100.00 2.79-2.91 0.00 100.00 2.91-3.03 0.00 100.00 3.03-3.14 0.00 100.00 3.14-3.26 0.00 100.00 3.26-3.38 0.00 100.00 3.38-3.50 0.00 100.00 3.50-3.62 0.00 100.00 3.62-3.74 0.00 100.00 3.74-3.86 0.00 100.00 3.86-3.98 0.00 100.00 3.98-4.10 0.00 100.00	Size (μ m) distribution % accumulation % 4.10-4.22 0.00 100.00 4.22-4.33 0.00 100.00 4.33-4.45 0.00 100.00 4.45-4.57 0.00 100.00 4.57-4.69 0.00 100.00 4.69-4.81 0.00 100.00 4.81-4.93 0.00 100.00 4.93-5.05 0.00 100.00 5.05-5.17 0.00 100.00 5.17-5.29 0.00 100.00 5.29-5.41 0.00 100.00 5.41-5.52 0.00 100.00 5.52-5.64 0.00 100.00 5.64-5.76 0.00 100.00 5.76-5.88 0.00 100.00 5.88-6.00 0.00 100.00 6.00-0.00 0.00 0.00

Fig. 2 granule number scattergram (2)

TSM ultra-fine grain Particle Size Analyzer, Shanghai University of Science and Technology
			Shanghai Institute of Pharmaceutical Industry's sample name: YZHH date: 06-03-2002 time: 14:48:20 sequence number: 1 D32=0.5375 Dv05=0.554 Dv01=0.459 Dv09=0.624 specific surface area: 11.162
Size (μ m) distribution % accumulation % 0.05-0.17 0.00 0.00 0.17-0.29 0.10 0.10 0.29-0.41 3.04 3.14 0.41-0.53 30.79 33.92 0.53-0.65 61.94 95.86	Size (μ m) distribution % accumulation % 2.07-2.19 0.00 100.00 2.19-2.31 0.00 100.00 2.31-2.43 0.00 100.00 2.43-2.55 0.00 100.00 2.55-2.67 0.00 100.00	Size (μ m) distribution % accumulation % 4.10-4.22 0.00 100.00 4.22-4.33 0.00 100.00 4.33-4.45 0.00 100.00 4.45-4.57 0.00 100.00 4.57-4.69 0.00 100.00

0.65-0.76 4.14 100.00 0.76-0.88 0.00 100.00 0.88-1.00 0.00 100.00 1.00-1.12 0.00 100.00 1.12-1.24 0.00 100.00 1.24-1.36 0.00 100.00 0.36-1.48 0.00 100.00 1.48-1.60 0.00 100.00 1.60-1.72 0.00 100.00 1.72-1.84 0.00 100.00 1.84-1.95 0.00 100.00 1.95-2.07 0.00 100.00

2.67-2.79 0.00 100.00 2.79-2.91 0.00 100.00 2.91-3.03 0.00 100.00 3.03-3.14 0.00 100.00 3.14-3.26 0.00 100.00 3.26-3.38 0.00 100.00 3.38-3.50 0.00 100.00 3.50-3.62 0.00 100.00 3.62-3.74 0.00 100.00 3.74-3.86 0.00 100.00 3.86-3.98 0.00 100.00 3.98-4.10 0.00 100.00

4.69-4.81 0.00 100.00 4.81-4.93 0.00 100.00 4.93-5.05 0.00 100.00 5.05-5.17 0.00 100.00 5.17-5.29 0.00 100.00 5.29-5.41 0.00 100.00 5.41-5.52 0.00 100.00 5.52-5.64 0.00 100.00 5.64-5.76 0.00 100.00 5.76-5.88 0.00 100.00 5.88-6.00 0.00 100.00 6.00-0.00 0.00 0.00

Fig. 3 granule number scattergram (3)

TSM ultra-fine grain Particle Size Analyzer, Shanghai University of Science and Technology
			Shanghai Institute of Pharmaceutical Industry's sample name: YZHH date: 06-03-2002 time: 14:52:37 sequence number: 1 D32=0.5376 Dv05=0.554 Dv01=0.459 Dv09=0.624 specific surface area: 11.160
Size (μ m) distribution % accumulation % 0.05-0.17 0.00 0.00 0.17-0.29 0.10 0.10 0.29-0.41 3.04 3.14 0.41-0.53 30.79 33.92 0.53-0.65 61.94 95.86 0.65-0.76 4.14 100.00 0.76-0.88 0.00 100.00 0.88-1.00 0.00 100.00 1.00-1.12 0.00 100.00 1.12-1.24 0.00 100.00 1.24-1.36 0.00 100.00 0.36-1.48 0.00 100.00 1.48-1.60 0.00 100.00 1.60-1.72 0.00 100.00 1.72-1.84 0.00 100.00 1.84-1.95 0.00 100.00 1.95-2.07 0.00 100.00	Size (μ m) distribution % accumulation % 2.07-2.19 0.00 100.00 2.19-2.31 0.00 100.00 2.31-2.43 0.00 100.00 2.43-2.55 0.00 100.00 2.55-2.67 0.00 100.00 2.67-2.79 0.00 100.00 2.79-2.91 0.00 100.00 2.91-3.03 0.00 100.00 3.03-3.14 0.00 100.00 3.14-3.26 0.00 100.00 3.26-3.38 0.00 100.00 3.38-3.50 0.00 100.00 3.50-3.62 0.00 100.00 3.62-3.74 0.00 100.00 3.74-3.86 0.00 100.00 3.86-3.98 0.00 100.00 3.98-4.10 0.00 100.00	Size (μ m) distribution % accumulation % 4.10-4.22 0.00 100.00 4.22-4.33 0.00 100.00 4.33-4.45 0.00 100.00 4.45-4.57 0.00 100.00 4.57-4.69 0.00 100.00 4.69-4.81 0.00 100.00 4.81-4.93 0.00 100.00 4.93-5.05 0.00 100.00 5.05-5.17 0.00 100.00 5.17-5.29 0.00 100.00 5.29-5.41 0.00 100.00 5.41-5.52 0.00 100.00 5.52-5.64 0.00 100.00 5.64-5.76 0.00 100.00 5.76-5.88 0.00 100.00 5.88-6.00 0.00 100.00 6.00-0.00 0.00 0.00

The specific embodiment

Embodiment 1

1. write out a prescription:

Herba Solidaginis oil 10g

Injection phospholipid 15g

Injection Oleum Glycines 100g

Glycerol 25g

Water for injection adds to 1000ml

2. preparation method:

(1) Herba Solidaginis oil that takes by weighing recipe quantity is gone in the injection Oleum Glycines of recipe quantity, and mixing is oil phase;

(2) the injection phospholipid that takes by weighing recipe quantity is gone in the homogenizer, adds the glycerol of proper amount of water for injection and recipe quantity, starts stirring, and phospholipid is dissolved fully, is water;

(3) under high-speed stirred, oil phase is added aqueous phase with wire, treat that oil phase finishes, continue to stir 10 minutes, promptly get milky colostric fluid;

(4) colostrum is put in the loading hopper of two step homogenizers, machined, when treating in the pipeline no bubble, first low pressure is adjusted to 5-15kg/cm ²: after treating that low pressure is stable, with high voltage adjusting to 30-50kg/cm ², after the circulation for several times Emulsion is transferred in the liquid receiver continuously;

(5) sodium hydroxide solution with 0.1N is adjusted to emulsion about pH7, and reuse water for injection is diluted to 1000ml;

(6) emulsion filters with No. 3 sintered filter funnels;

(7) carrying out intermediate mass detects;

(8) with the emulsion embedding in proper container;

(9) emulsion after the embedding is through 100 ℃ of flowing steam sterilizations 30 minutes, promptly.

3. result:

(1) content of effective

(2) pH value: 5.0-7.5

(3) size and the scope of breast grain: on average below 1 μ m, the result as shown in Figure 1 for the size of the breast grain of this product.

Embodiment 2

1. write out a prescription:

Herba Solidaginis oil 50g

Injection phospholipid 20g

Injection Oleum Glycines 150g

Glycerol 25g

Water for injection adds to 1000ml

2. preparation method: with embodiment 1

3. result:

(1) content of effective:

(2) pH value: 5.0-7.5

(3) size and the scope of breast grain: on average below 1 μ m, the result as shown in Figure 2 for the size of the breast grain of this product.

Embodiment 3

1. write out a prescription:

Herba Solidaginis oil 80g

Injection phospholipid 20g

Injection Oleum Glycines 200g

Glycerol 25g

Water for injection adds to 1000ml

2. preparation method: with embodiment 1

3. result:

(1) content of effective:

(2) pH value: 5.0-7.5

(3) size and the scope of breast grain: on average below 1 μ m, the result as shown in Figure 3 for the size of the breast grain of this product.

Claims (3)

1. fatty emulsion of goldenrod for injection is characterized in that this Emulsion is that active ingredient and pharmaceutic adjuvant are formed with the Herba Solidaginis oil, comprising:

Herba Solidaginis oil 1.0-100g

Injection phospholipid 3-50g

Injection Oleum Glycines 50-300g

Glycerol 25g

Water for injection adds to 1000ml

This product active component is a Herba Solidaginis oil, and concentration is 0.1-10%; Adjuvant is an injection Oleum Glycines, and concentration is 5-30% (W/W); Injection phospholipid, concentration are 0.3-5% (W/W); Glycerol and water for injection; Wherein, injection Oleum Glycines is as oil phase, and injection phospholipid is as surfactant, and glycerol is as isotonic agent, and water for injection is as water.

2. the preparation method of a fatty emulsion of goldenrod for injection as claimed in claim 1 is characterized in that this method comprises the following steps:

1. take by weighing above-mentioned Herba Solidaginis oil and go in the injection Oleum Glycines, mixing is oil phase;

2. take by weighing injection phospholipid and go in the homogenizer, add proper amount of water for injection and glycerol, start stirring, phospholipid is dissolved fully, be water;

3. under high-speed stirred, oil phase is added aqueous phase with wire, treat that oil phase finishes, continue to stir 10 minutes, promptly get milky colostric fluid;

4. colostrum is put in the loading hopper of two step homogenizers, machined, when treating in the pipeline no bubble, first low pressure is adjusted to 5-15kg/cm ²After treating that low pressure is stable, with high voltage adjusting to 30-50kg/cm ², after the circulation for several times Emulsion is transferred in the liquid receiver continuously;

5. the sodium hydroxide solution with 0.1N is adjusted to emulsion about pH7, and reuse water for injection is diluted to 1000ml;

6. emulsion filters with No. 3 sintered filter funnels;

7. carrying out intermediate mass detects;

8. with the emulsion embedding in proper container;

9. the emulsion after the embedding is through 100 ℃ of flowing steam sterilizations 30 minutes, promptly.

A Herba Solidaginis as claimed in claim 1 have in preparation that analgesia is analgesic, the application in the fat emulsion for injection of antiinflammatory and adjusting immunologic function.

Images