Summary of the invention
The object of the invention is, a kind of preparation method of eyebrow generating liquid is provided, this method is to utilize membrane technology further to develop the extract of water on the basis of Chinese patent 921148089, be that Folium Isatidis, Semen Nigellae, Radix Polygoni Multiflori, Cacumen Platycladi, Fructus Chebulae are mixed, add water, extract merge extractive liquid,, standing over night gets supernatant; Thin up again carries out ultrafiltration with the hollow-fibre membrane of composite polypropylene in room temperature, merges ultrafiltrate, the reuse NF membrane concentrates, and in concentrated solution, adds ultramicro active charcoal, HANSHENGJIAO powder, glycerol, antiseptic, essence successively, mix, can obtain eyebrow generating liquid.The eyebrow generating liquid that obtains by this method can solve the eyebrow rareness, fall the demand of eyebrow and the vast colony of scarce eyebrow, satisfies people to pursuit beautiful and high spiritual quality life.Thereby solve instability, very easily variable color and stratified problem in the environment of southern high-temperature humidity.This eyebrow generating liquid is the liquid of blackish green, has the unique fragrance abnormal smells from the patient.
The preparation method of a kind of eyebrow generating liquid of the present invention follows these steps to carry out:
A, Folium Isatidis 250-500g, Semen Nigellae 100-300g, Radix Polygoni Multiflori 150-400g, Cacumen Platycladi 150-400g, Fructus Chebulae 100-300g are ground into fine powder mix, add the water of 8 times of amounts, extract 3 times, each 0.5-1.5 hour, merge extractive liquid,, standing over night gets supernatant;
B, add the water dilution that 3-6 doubly measures again, carry out ultrafiltration in pressure 0.06-0.12MPa room temperature, repeat 2-5 time with the hollow-fibre membrane of composite polypropylene, the merging ultrafiltrate, the reuse NF membrane concentrates;
C, with in the step b concentrated solution, add ultramicro active charcoal 7-12g, HANSHENGJIAO powder 1-3g, glycerol 8-12ml, antiseptic 15-30ml, essence 0.2-0.8ml successively, stirred 24-48 hour, can obtain eyebrow generating liquid.
The hollow-fibre membrane specification of step b composite polypropylene is 6000-50000.
The volume ratio of step b NF membrane concentrated solution and crude drug is 1: 1.
The HANSHENGJIAO powder expands with 50ml is water-soluble among the step c.
The preparation method of a kind of eyebrow generating liquid of the present invention, the animal experiment that the eyebrow generating liquid that obtains by this method has carried out the application of living eyebrow:
Experiment material
Laboratory animal
Inbred line C57BL/6 mice (whole body is a black hair mice), female, in 6~8 ages in week, body weight 15~20g is bought by Test Animal Centre, Academy of Military Medical Sciences, P.L.A, animal credit number: SCXK-(army) 2007-004.Select the mice that skin is pink, hair is in resting stage.
Tried thing
Administration group (eyebrow generating liquid): eyebrow generating liquid source: by the eyebrow generating liquid of the inventive method acquisition.
Matched group (distilled water): distilled water.
Positive controls (ZHANGGUANG 101): buy commercially available hair growth promoting commodity (ZHANGGUANG101 YUFAJI, lot number 20080701, BeiJing ZhangGuang's 101 development in science and technology company limiteies).
Reagent
Sodium sulfide: three subsidiary factories of Tianjin chemical reagent six factories, lot number 20061106;
Formaldehyde: Xi'an chemical reagent factory, lot number 070530.
Test method:
Inducing of trophophase
With 8% sodium sulfide solution depilation, the unhairing area is about 4cm to preceding 24 hours of administration with mouse back
2, wipe the hair that comes off with wet cloth after 2 minutes, guarantee that local hair is taken off fully and not damaged.
Animal grouping and administration
Get 24 of depilation mices, be divided into three groups of matched group (distilled water), positive controls (ZHANGGUANG 101) and administration groups (eyebrow generating liquid) at random, every group 8, each group is all directly smeared matched group (distilled water), positive controls (ZHANGGUANG 101) and administration group (eyebrow generating liquid) at the depilation position, smear at regular time and quantity once every day, smear about 0.20ml, observe every day, continuous 23 days at every turn.
Observation index
Observe the mice color of the leather every day and change, respectively record:
The time of every mice depilation district skin color from pink to the beginning blackening (my god);
Skin begin blackening to hair grow skin surface time (my god).
Histopathologic examination
After educating hair experiment end, put to death mice with disconnected vertebra method;
The position of smearing at each group mouse back cuts skin sample;
Pathology slice probing method is fixed with 10% formalin routinely, dehydration, specimens paraffin embedding slices, hematoxylin-eosin staining;
The histology that light microscopic is observed the mouse skin hair follicle down changes, and counting high dermis hair follicle number.
Statistical procedures
Experimental data is with mean ± standard deviation (x ± S) expression, t check between the employing group.
The result:
Mouse back depilation district skin color changes
Matched group (distilled water) mice becomes black (promptly entering trophophase by resting stage) in the 9th to the 10 day skin in depilation back by pink, the 14th day hair in depilation back grows the skin table, part mouse skin color became grey black (promptly enter catagen by trophophase, promptly hair growth is slow) in the 23rd day;
Positive controls (ZHANGGUANG 101) mice becomes black (promptly entering trophophase by resting stage) in the 7th to the 8 day skin in depilation back by pink, and the 10th day hair in depilation back grows the skin table, and the 23rd day mouse skin still is black (trophophase, promptly hair continues growth).
Administration group (eyebrow generating liquid) mice becomes black (promptly entering trophophase by resting stage) in the 6th to the 7 day skin in depilation back by pink, and the 9th day hair in depilation back grows the skin table, and the 23rd day mouse skin still is black (trophophase, promptly hair continues growth).
Statistical result shows:
Administration group (eyebrow generating liquid) compares with matched group (distilled water), but administration group highly significant shortens the time (P<0.01) of C57BL/6 mice depilation back skin color blackening;
Administration group (eyebrow generating liquid) compares with matched group (distilled water), and administration group (eyebrow generating liquid) can significantly shorten skin and begin blackening grows epidermis to hair time (P<0.05).Experimental result sees Table 1.
Table 1 eyebrow generating liquid is to the influence of the variation of mouse back depilation district skin color (x ± SD)
Annotate: administration group (eyebrow generating liquid) compares with matched group (distilled water), * * P<0.01 * P<0.05,
Histopathologic examination:
Histopathological examination is the result show,
Administration group (eyebrow generating liquid), matched group (distilled water) and positive controls (ZHANGGUANG 101) mouse back skin texture are all complete, and epidermis is not seen necrosis;
Corium is not seen cell infiltration;
Each group is all seen the normal hair follicle of structure;
With 100 times of visual field calculating numbers, administration group (eyebrow generating liquid) and positive controls (ZHANGGUANG 101) hair follicle digital display work are more than matched group (distilled water).
Statistical result shows:
Administration group (eyebrow generating liquid) compares with matched group (distilled water), and administration group (eyebrow generating liquid) high dermis hair follicle number obviously increases, and (P<0.05) sees Table 2.
Table 2 eyebrow generating liquid is to mouse back test block skin follicle counting (x ± SD)
Annotate: administration group (eyebrow generating liquid) compares with matched group (distilled water), and administration group (eyebrow generating liquid) hair follicle digital display work is more than matched group (distilled water) * P<0.05; Administration group (eyebrow generating liquid) does not have significant difference with positive controls (ZHANGGUANG 101) hair follicle number.
The C57BL/6 mice educates a mao back dermatopathology and observes
Experiment conclusion:
Conclusion
Administration group (eyebrow generating liquid) is to C57BL/6 mice hair growth influence experiment, and the result shows:
Administration group (eyebrow generating liquid) compares with matched group (distilled water), the time (promptly promoting the generation of trophophase melanocyte) of administration group (eyebrow generating liquid) back skin color blackening but the shortening C57BL/6 mice of highly significant loses hair or feathers;
Administration group (eyebrow generating liquid) also can significantly shorten skin and begin blackening grows skin surface to hair time.Confirm that administration group (eyebrow generating liquid) has remarkable trichogenous effect;
Histopathological examination is the result show, administration group (eyebrow generating liquid) compares with matched group (distilled water), and the mouse back skin texture is complete; Epidermis is not seen necrosis; Corium is not seen cell infiltration; Administration group and matched group are all seen the normal hair follicle of structure;
Administration group (eyebrow generating liquid) hair follicle is obviously more than matched group (distilled water).
Prompting administration group (eyebrow generating liquid) is safe and effective to dermatologic.
Mice educates after hair experiment finishes, and disconnected vertebra is put to death and got butt, and butt tissue shows that through the pathological examination result administration group (eyebrow generating liquid) compares with matched group (distilled water) and positive controls (ZHANGGUANG 101):
The mouse back skin texture is complete, and epidermis is not seen necrosis;
Corium is not seen cell infiltration;
The normal hair follicle of all high-visible structure;
Administration group (eyebrow generating liquid) hair follicle is obviously more than matched group (distilled water).
Prompting administration group (eyebrow generating liquid) to dermatologic safety and hair follicle obviously more than other groups.
Give birth to the clinical trial that eyebrow is used by the eyebrow generating liquid that the method for the invention obtains as preparation:
Experiment material
The experimenter
The volunteer that eyebrow is sparse
Tried thing
Eyebrow generating liquid obtains by the method for the invention.
Test method:
Volunteer 10 people that eyebrow is sparse, after clean every night, rub with finger and smeared 2 times along the direction of eyebrow by the reagent product after supercilium 1-2 minute, operating period is 23 days, observe the variation of length, density and the color and luster of medication front and back eyebrow, length, density and color and luster to eyebrow are marked, and estimate with 0 minute (poor), 2 minutes (generally), 4 minutes (better) and 6 minutes (fine) respectively.
Statistical procedures
Experimental data is with mean ± standard deviation (x ± S) expression, t check in the employing group.
Table 3 eyebrow generating liquid is to the influence of eyebrow growth
Annotate: administration (eyebrow generating liquid) after 23 days with administration (eyebrow generating liquid) 0 day relatively,
*P<0.05;**P<0.01。
Tried the evaluation of thing effectiveness
According to being tried the scoring of thing, tried the back 23 days total scoring T of each experimenter to eyebrow length, density and color and luster
nDeduct 0 day total scoring T
0(T=T
n-T
0) 1. T≤2 be invalid; 2. 2<T≤6 are effective; 3. T>6 are healing, effective percentage %=(effectively+cure)/10 * 100%, invalid 3 examples of result, effective 4 examples are cured 3 examples, and effective percentage is 70%.
Experiment conclusion
Administration (eyebrow generating liquid) after 23 days with administration (eyebrow generating liquid) 0 day relatively, administration (eyebrow generating liquid) after 23 days eyebrow length significantly more than 0 day * P<0.05 of administration (eyebrow generating liquid); Administration (eyebrow generating liquid) after 23 days with administration (eyebrow generating liquid) 0 day relatively, have to promote eyebrow density to increase very significantly and the effect of color and luster blackening.**P<0.01。
Administration (eyebrow generating liquid) is after 23 days, is 70% to the total effective rate of eyebrow length, density and color and luster.
The specific embodiment
Embodiment 1
A, Folium Isatidis 250g, Semen Nigellae 300g, Radix Polygoni Multiflori 400g, Cacumen Platycladi 400g, Fructus Chebulae 300g are ground into fine powder mix, add the water of 8 times of amounts, extracts 3 times, each 1.5 hours, merge extractive liquid,, standing over night must supernatant;
B, adding the water dilution of 3 times of amounts again, is that 6000 hollow-fibre membrane carries out ultrafiltration in pressure 0.06MPa room temperature with the composite polypropylene specification, repeat 2 times, and the merging ultrafiltrate, the reuse NF membrane concentrates, and the volume ratio of concentrated solution and crude drug is 1: 1
C, with in the step b concentrated solution, add successively ultramicro active charcoal 7g, HANSHENGJIAO powder 1g with 50ml water-solublely expand, glycerol 8ml, antiseptic 15ml, essence 0.2ml, stirred 24 hours, can obtain eyebrow generating liquid, this eyebrow generating liquid is the liquid of blackish green, has the unique fragrance abnormal smells from the patient
Embodiment 2
A, Folium Isatidis 350g, Semen Nigellae 150g, Radix Polygoni Multiflori 250g, Cacumen Platycladi 250g, Fructus Chebulae 150g are ground into fine powder mix, add the water of 8 times of amounts, extracts 3 times, each 1.0 hours, merge extractive liquid,, standing over night must supernatant;
B, adding the water dilution of 4 times of amounts again, is that 10000 hollow-fibre membrane carries out ultrafiltration in pressure 0.08MPa room temperature with the composite polypropylene specification, repeat 3 times, and the merging ultrafiltrate, the reuse NF membrane concentrates, and the volume ratio of concentrated solution and crude drug is 1: 1;
C, with in the step b concentrated solution, add successively ultramicro active charcoal 9g, HANSHENGJIAO powder 2g with 50ml water-solublely expand, glycerol 10ml, antiseptic 20ml, essence 0.4ml, stirred 30 hours, can obtain eyebrow generating liquid, this eyebrow generating liquid is the liquid of blackish green, has the unique fragrance abnormal smells from the patient.
Embodiment 3
A, Folium Isatidis 500g, Semen Nigellae 100g, Radix Polygoni Multiflori 150g, Cacumen Platycladi 150g, Fructus Chebulae 100g are ground into fine powder mix, add the water of 8 times of amounts, extracts 3 times, each 0.5 hour, merge extractive liquid,, standing over night must supernatant;
B, adding 6 times water dilution again, is that 50000 hollow-fibre membrane carries out ultrafiltration in pressure 0.12MPa room temperature with the composite polypropylene specification, repeat 4 times, and the merging ultrafiltrate, the reuse NF membrane concentrates, and the volume ratio of concentrated solution and crude drug is 1: 1;
C, with in the step b concentrated solution, add successively ultramicro active charcoal 12g, HANSHENGJIAO powder 3g with 50ml water-solublely expand, glycerol 10ml, antiseptic 30ml, essence 0.8ml, stirred 48 hours, can obtain eyebrow generating liquid, this eyebrow generating liquid is the liquid of blackish green, has the unique fragrance abnormal smells from the patient.
Embodiment 4
A, Folium Isatidis 430g, Semen Nigellae 250g, Radix Polygoni Multiflori 350g, Cacumen Platycladi 350g, Fructus Chebulae 250g are ground into fine powder mix, add the water of 8 times of amounts, extracts 3 times, each 1.2 hours, merge extractive liquid,, standing over night must supernatant;
B, adding the water dilution of 5 times of amounts again, is that 30000 hollow-fibre membrane carries out ultrafiltration in pressure 0.10MPa room temperature with the composite polypropylene specification, repeat 4 times, and the merging ultrafiltrate, the reuse NF membrane concentrates, and the volume ratio of concentrated solution and crude drug is 1: 1;
C, with in the step b concentrated solution, add successively ultramicro active charcoal 10g, HANSHENGJIAO powder 2.5g with 50ml water-solublely expand, glycerol 12ml, antiseptic 25ml, essence 0.6ml, stirred 36 hours, can obtain eyebrow generating liquid, this eyebrow generating liquid is the liquid of blackish green, has the unique fragrance abnormal smells from the patient.