CN109718258A - A kind of gumbo eye drops and preparation method thereof - Google Patents
A kind of gumbo eye drops and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a kind of gumbo eye drops and preparation method thereof.The gumbo eye drops includes following components: gumbo extract and solvent；The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-96%；The mass percentage that the solvent accounts for the gumbo eye drops is 4%-96%；The solvent is water or PBS.Gumbo eye drops of the invention has preferable eye of alleviating dry and astringent, tired, treats the effect of xerophthalmia, and irritation is small, price is suitable, and gumbo is resourceful, and the preparation process of gumbo eye drops is simple, has preferable application and popularization value.
Description
Technical field
The present invention relates to a kind of gumbo eye drops and preparation method thereof.
Background technique
In recent years, with universal, the intensity of people's eye of the increase of Air haze, smart phone and entertainment electronics appliances
It obviously increasing, eye health situation is also remarkably decreased, and more and more people are easy to feel that eyes are dry and astringent, tired, and it is uncomfortable, into
And generate xerophthalmia.The method of conventional treatment xerophthalmia is the treatment or alleviation xerophthalmia using eye drops, and presently commercially available
Eye drops in, there is a problem of that more such as expensive, effect is unobvious, some products are even there are also certain stimulation
Property, therefore develop new eye drops and have great importance.
Summary of the invention
The problems such as expensive, ineffective present invention aim to address the presently commercially available eye drops in this field, in turn
Provide a kind of gumbo eye drops and preparation method thereof.
Gumbo is Malvaceae Abelmoschus annual herb plant also known as okra, swordweed, purling eggplant, Abelmoschus esculentus, is one
Kind low fat, integration of drinking and medicinal herbs vegetables low in calories, without cholesterol.The present inventor is by the discovery of long-term observational study, autumn
Certain herbaceous plants with big flowers species resource is enriched, has a very wide distribution, and plant resources are easy to obtain, cheap, and on this basis, inventor is developed into
A kind of gumbo eye drops of function.The gumbo eye drops has the effect of preferably alleviating eye dry and astringent, tired, and irritation
Small, price is suitable, preparation is simple, has preferable application and popularization value.
The present invention is to solve technical problem by the following technical programs:
The present invention provides a kind of gumbo eye drops, wherein the gumbo eye drops includes following components: gumbo is extracted
Object and solvent；
The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-96%；
The mass percentage that the solvent accounts for the gumbo eye drops is 4%-96%；
The solvent is water or PBS；
The gumbo extract is made by following preparation method, the preparation method the following steps are included:
(1) gumbo pod is cleaned, chopping, homogenate, carries out water extraction, obtains crude extract；
(2) the crude extract ethanol precipitation for obtaining step (1) is centrifuged, dry, obtains jelly；
(3) jelly for obtaining step (2) dissolves, and dialyses in the distilled water of 15-20 times of volume, is freeze-dried, i.e.,
It can.
The gumbo eye drops, preferably, it is made of gumbo extract and solvent；Wherein,
The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-96%；
The mass percentage that the solvent accounts for the gumbo eye drops is 4%-96%；
The solvent is water or PBS.
The gumbo eye drops, preferably, including the following components: gumbo extract, solvent and auxiliary material；Wherein,
The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-95%；
The mass percentage that the solvent accounts for the gumbo eye drops is 4%-95%；
The mass percentage that the auxiliary material accounts for the gumbo eye drops is 0.1%-1%；
The auxiliary material includes preservative.
Wherein, the preservative is better selected from one of sodium benzoate, sorbic acid and ethyl hydroxy benzoate or a variety of, more preferably
Ground is sodium benzoate.
The gumbo eye drops, wherein
Gumbo pod described in step (1) is gumbo fruit, preferably the fresh pod of gumbo.
The operation that water described in step (1) extracts can be this field routine water extraction operation, be preferably comprised following steps:
The distilled water of gumbo pod and 5-10 times of volume after homogenate is mixed, stands 12-24 hour, centrifugation, removing sediment to get
The crude extract；The revolving speed of the centrifugation can be 2000-4000rpm, and the time of centrifugation can be 2-5min.
The mass percent concentration of ethyl alcohol described in step (2) can be 30%-40%, be more preferably 35%.
In step (2), the revolving speed of the centrifugation can be 2000-3500rpm, and the time of centrifugation can be 3-10min.
In step (2), the drying can be this field conventional drying methods, preferably be dried in vacuo, and the vacuum is dry
Dry temperature is preferably 50-60 DEG C.
The operation of dissolution described in step (3) can be this field routinely dissolution operation, preferably be dissolved in jelly
In the distilled water of a small amount of volume, wherein a small amount of volume is not specially limited, as long as the jelly can be dissolved.
In step (3), the method for the dialysis can be this field conventional dialysis procedures, preferably be carried out using dialysis membrane,
The dialysis membrane is preferably MD44 dialysis membrane, and by American Association, Carbide is provided.The time of the dialysis is preferably
24-48 hours；The dialysis further includes the operation that the liquid outside dialysis membrane is replaced using new distilled water, time of the replacement
Number preferably daily 3-4 times.
The gumbo eye drops, wherein
The PBS is phosphate buffered saline solution, and PBS of the present invention can be this field routine PBS, and preferably its pH is
7.2-7.4。
The water is preferably distilled water.
The pH value of the gumbo eye drops can be 4.60-4.70, be more preferably 4.65.
The gumbo extract accounts for the mass percentage preferred 5%-95% of the gumbo eye drops, more preferable 25% or
50%.
Gumbo of the present invention is Malvaceae Abelmoschus gumbo Hibiscus esculentus L..
The present invention also provides the preparation methods of the gumbo eye drops, wherein the preparation method comprises the following steps:
The gumbo extract and solvent are mixed.
The preparation method of the gumbo eye drops, wherein when the gumbo eye drops contains auxiliary material, preparation method
The following steps are included: by gumbo extract, solvent and the auxiliary materials and mixing.
In the present invention, the gumbo eye drops is for alleviating or treating xerophthalmia, treatment or application method are as follows: it is straight
Dropwise addition is connect in user's eyelid, 1-2 drop is added dropwise every time, is added dropwise 3-4 times daily.
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can any combination to get the present invention it is each preferably
Example.
The reagents and materials used in the present invention are commercially available.
The positive effect of the present invention is that:
Gumbo eye drops provided by the invention has preferable eye of alleviating dry and astringent, tired, treats the effect of xerophthalmia, pierces
Sharp property is small, price is suitable, and gumbo is resourceful, and the preparation process of gumbo eye drops is simple, has preferable application valence
Value.
Detailed description of the invention
Fig. 1 is the therapeutic effect of gumbo eye drops 3 in effect example 3 of the present invention, and wherein A is phenol red staple test knot
Fruit, B are BUT test result, and C is FL test result.
Fig. 2 is in effect example 3 of the present invention, and gumbo eye drops 3 treats the BUT test result of rat, and wherein A is treatment
Before, after B is treatment 1 week, after C is treatment 2 weeks.
Specific embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to the reality
It applies among a range.In the following examples, the experimental methods for specific conditions are not specified, according to conventional methods and conditions, or according to quotient
The selection of product specification.
Instrument used in the embodiment of the present invention include the following:
DT5-1B low speed centrifuge (Beijing epoch Bei Li centrifuge Co., Ltd)；
RE-52 Rotary Evaporators (Shanghai Yarong Biochemical Instrument Plant)；
TE1245 electronic balance (Beijing Sai Duolisi instrument system Co., Ltd)；
ALPHA2-4 freeze drying equipment (German CHRIST company)；
Dialysis membrane is MD44 film (American Association Carbide)；
PBS provides for Thermo Fischer Scient Inc..
Other instruments and reagent are commercially available.
The gumbo that the embodiment of the present invention uses originates from industry, is Malvaceae Abelmoschus gumbo Hibiscus
Esculentus L., after fresh picking clean dry it is spare.
Prepare the preparation of 1 gumbo extract of embodiment
Take 1 kilogram clean after gumbo pod, shred, homogenate, be added after 3 liters of distilled water stand 24 hours and be centrifuged, be centrifuged
Revolving speed be 3000rpm, be centrifuged 5 minutes, remove sediment, obtain 0.96 kilogram of crude extract.
The ethyl alcohol of 35% concentration is slowly added into crude extract, it is stirring while adding, it precipitates, centrifugation, the revolving speed of centrifugation is
3200rpm is centrifuged 4 minutes, collects sediment, sediment is dried in vacuo, 109 jellies are obtained.
Jelly is dissolved in 500 milliliters of water, is dialysed 48 hours in the distilled water of 20 times of volumes, wherein changing liquid daily
It 4 times, is freeze-dried after dialysis, obtains 35 grams of gumbo extracts.
Prepare the preparation of embodiment 2-5 gumbo eye drops
Gumbo eye drops of the present invention the preparation method comprises the following steps: gumbo extract and PBS are mixed.The wherein autumn
Certain herbaceous plants with big flowers extract is prepared in present invention preparation embodiment 1.The composition of the gumbo eye drops of preparation is according to described in following table.
Table 1
The effect expedition of gumbo eye drops in treatment xerophthalmia
What effect example of the invention was investigated is the effect of gumbo eye drops in treatment xerophthalmia.
Zoopery is as follows in the embodiment of the present invention:
1, experimental animal
The female Wistar rats (weight is about 180-210g, 10-12 week old) for selecting ocular and whole body health, by Shanghai
This Lake experimental animal Co., Ltd provides.Eyes ocular and anterior ocular segment inspection under slit-lamp are carried out to rat before experiment,
As a result normal.It is placed in standard sterile environment, rat feeding condition are as follows: room temperature (20-25 DEG C), relative humidity (50-60%), lamp
Optical analog day and night change (light application time 8:00-22:00).Clean feed is provided by animal center, and food and water supply are sufficient.Institute
There are zoopery and experiment condition to strictly observe " management of laboratory animal regulations ".
2, experimentation
2.1 dry eye models are established
Experimental rat eyes are dripped with 0.2% benzalkonium chloride eye drops, and 1 day 3 times, totally 1 week, thus induced rat dry eyes mould
Type.
2.2 evaluation index
2.2.1 breakup time of tear film (BUT)
0.1% Fluress drop is in rat conjunctival sac, after artificial auxiliary is twinkled 3 times, with slit-lamp cobalt blue light into
Row observation.Fissura palpebrae open to first breakdown point of tear film occur when a length of breakup time of tear film as a result, to judge tear film
Stability.
2.2.2 lacrimal secretion detection (phenol red staple test)
One end of phenol red staple is placed at rat lower tarsal conjunctiva capsule China and foreign countries 1/3, cotton thread reddens part after placing 20 seconds
Length is denoted as lacrimal secretion test result, thus the result can reflect ocular lacrimal secretion as unit of mm/20s.
2.2.3 corneal fluorescein dyeing (FL)
By 0.1% Fluress drop after rat conjunctival sac, observed under slit-lamp cobalt blue light.As a result can reflect
The integrality of skin contaminates display corneal epithelial defect position and range by fluorescence.
Standards of grading: 4 quadrants are classified as from corneal center, each quadrant score, 16 points are total score.0 point: without dotted
Dyeing；1 point: a small amount of dotted dyeing, but it is less than 30 points；2 points: dotted dyeing is greater than 30, but not disperse；3 points: dispersivity point
Shape dyeing, but dyed without patch shape；4 points: the dyeing of patch shape.
Effect example 1
What this effect example was investigated is (the gumbo extractive content of gumbo eye drops 1 prepared by present invention preparation embodiment 2
For the effect for 5%) treating dry eyes.Wherein the test method of phenol red staple, BUT and FL are as described above, test result is shown in Table 2.
Table 2
In this effect example, 5 rats totally 10 eyes are shared, average phenol red staple test result and tear before treatment
Film rupture time result is respectively 8.00 ± 1.22,2.07 ± 0.50.
From table 2 it can be seen that dry eyes inspection result makes moderate progress earlier above, there is statistics after gumbo eye drops 1 is treated 1 week
Meaning.After treating 2 weeks, phenol red staple test result is 11.99 ± 2.10, breakup time of tear film 3.14 ± 0.67, the two point
Not compared with before treatment, difference is statistically significant.
FL test: cornea of rats scoring is fallen to from 11.00 ± 0.99 before treatment after gumbo eye drops 1 is treated 1 week
9.14±0.70；After treating 2 weeks, the corneal dyeing scoring before cornea of rats dyeing scoring about 8.05 ± 0.75, with treatment has
Obvious statistical difference.Cornea of rats epithelium has diffused dotted dye before treating, and the fluorescent staining after treatment 2 weeks is bright earlier above
It is aobvious to improve.
Effect example 2
What this effect example was investigated is (the gumbo extractive content of gumbo eye drops 2 prepared by present invention preparation embodiment 3
For the effect for 25%) treating dry eyes.Wherein the test method of phenol red staple, BUT and FL are as described above, test result is shown in Table 3.
Table 3
In this effect example, 5 rats totally 10 eyes are shared, average phenol red staple test result and tear before treatment
Film rupture time result is respectively 8.93 ± 1.29,2.57 ± 0.70.
After gumbo eye drops 2 is treated 1 week, dry eyes inspection result makes moderate progress earlier above, but difference and not statistically significant.It controls
After treating 2 weeks, phenol red staple test result be 12.99 ± 2.20, breakup time of tear film 3.04 ± 0.66, the two respectively with treatment
Preceding comparison, difference are statistically significant.
FL test: cornea of rats scoring is fallen to from 10.55 ± 1.07 before treatment after gumbo eye drops 2 is treated 1 week
After 9.04 ± 0.75, treatment 2 weeks, the corneal dyeing scoring before scoring about 8.75 ± 0.70, with treatment has obvious statistics poor
It is different.Cornea of rats epithelium has diffused dotted dye before treating, and the fluorescent staining after treatment 2 weeks is clearly better earlier above.
Effect example 3
What this effect example was investigated is (the gumbo extractive content of gumbo eye drops 3 prepared by present invention preparation embodiment 4
For the effect for 50%) treating dry eyes.Wherein the test method of phenol red staple, BUT and FL are as described above, test result is shown in Table 4.
In this effect example, 5 rats totally 10 eyes are shared.Phenol red staple before the treatment of 50% gumbo eye drops group is surveyed
Test result and breakup time of tear film are respectively 9.33 ± 1.39；After 2.67 ± 0.80, treatment 1 week, test result is respectively 9.08
± 1.11,2.75 ± 0.83, it is clearly better before relatively treating.After treating the 2nd week, test result is respectively 13.9 ± 2.40,3.54
± 0.66 is preceding with obvious statistical significance with treatment.
Corneal dyeing: cornea of rats average score is 10.42 ± 1.27 before treating, and 1,2 after gumbo eye drops in treatment
Zhou Hou, scoring are respectively 10.04 ± 0.78,8.75 ± 0.75, are clearly better before relatively treating.Have after treating the 2nd week obvious
Statistical significance.
Table 4
In this effect example, cornea of rats BUT the result is shown in Figure 1, rat BUT observation chart is shown in Fig. 2.Breakup time of tear film two
For person respectively compared with before treatment, difference is statistically significant.
Effect example 4
What this effect example was investigated is (the gumbo extractive content of gumbo eye drops 4 prepared by present invention preparation embodiment 5
For the effect for 95%) treating dry eyes.Wherein the test method of phenol red staple, BUT and FL are as described above, test result is shown in Table 5.
Table 5
In this effect example, 5 rats totally 10 eyes, the phenol red staple test result average out to before rat treatment are shared
4.6 ± 1.77, breakup time of tear film is 1.4 ± 0.37.
After gumbo eye drops 4 is treated 1 week, test result is respectively 7.8 ± 1.33,3.2 ± 0.51, obvious before relatively treating
It improves.After treating 2 weeks, the phenol red staple result of rat rises to 12.8 ± 0.75, BUT and rises to 3.50 ± 0.60, before treatment
And the difference for the treatment of 1 week has obvious statistical significance.
FL test: cornea of rats average score is 11.4 ± 1.36 before treating, and is treated 1 week and 2 weeks through gumbo eye drops 4
Afterwards, scoring is respectively 7.8 ± 1.78,7 ± 1.09, is clearly better before relatively treating.
Contrast effect embodiment 1
What this contrast effect embodiment 1 was investigated is (95% gumbo of gumbo eye drops 4 prepared by present invention preparation embodiment 5
Eye drops) and paired observation of the PBS eye drops in the dry eye model that Wistar rat is induced.
1, experimental animal
The female rats (weight is about 180-210g, 10-12 week old) for selecting ocular and whole body health, by Shanghai Si Laike
Experimental animal Co., Ltd provides.Eyes ocular and anterior ocular segment inspection under slit-lamp are carried out to rat before experiment, as a result
Normally.It is placed in standard sterile environment, rat feeding condition are as follows: room temperature (20-25 DEG C), relative humidity (50-60%), light simulation
Day and night change (light application time 8:00-22:00).Clean feed is provided by animal center, and food and water supply are sufficient.Progress institute
There are zoopery and experiment condition to strictly observe " management of laboratory animal regulations ".
2, experimentation
2.1 dry eye models are established
Totally 15 Wistar rats are randomly divided into 95% gumbo eye drops group and PBS group.Eyes are dripped to be pricked with 0.2% benzene
Oronain eye drops 1 day 3 times, totally 1 week, induces Wistar rat dry eye model.
2.2 evaluation index
2.2.1 breakup time of tear film (BUT)
0.1% Fluress drop is in rat conjunctival sac, after artificial auxiliary is twinkled 3 times, with slit-lamp cobalt blue light into
Row observation.Fissura palpebrae open to first breakdown point of tear film occur when a length of breakup time of tear film as a result, to judge tear film
Stability.
2.2.2 lacrimal secretion detection (phenol red staple test)
One end of phenol red staple is placed at rat lower tarsal conjunctiva China and foreign countries 1/3, cotton thread reddens the length of part after placing 20 seconds
Degree is denoted as lacrimal secretion test result, can reflect ocular lacrimal secretion.
2.2.3 corneal fluorescein dyeing (FL)
By 0.1% Fluress drop after in rat conjunctival sac, observed under slit-lamp cobalt blue light.As a result can reflect
The integrality of epithelium contaminates display corneal epithelial defect position and range by fluorescence.
Standards of grading: 4 quadrants are classified as from corneal center, each quadrant score, 16 points are total score.0 point: without dotted
Dyeing；1 point: a small amount of dotted dyeing, but it is less than 30 points；2 points: dotted dyeing is greater than 30, but not disperse；3 points: dispersivity point
Shape dyeing, but dyed without patch shape；4 points: the dyeing of patch shape.
3, result
Dry eyes inspection: tear film rupture and phenol red staple test result before the treatment of 95% gumbo eye drops group Wistar rat
With treatment 1,2 week after result have obvious statistical difference.In addition, 95% gumbo eye drops group and PBS group are treating 2 Zhou Houqi tears
Film rupture time is clearly better with phenol red staple test result.
Corneal dyeing: after 95% gumbo eye drops group is treated 1,2 week, scoring is clearly better before relatively treating.This group of angle
Film dyeing scoring has significant difference with the scoring of PBS group after treatment 1,2 week, and Dry eye treatment group effect is preferable.Test result is shown in
Table 6.
Table 6
Claims (10)
1. a kind of gumbo eye drops, which is characterized in that the gumbo eye drops includes following components: gumbo extract and molten
Agent；
The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-96%；
The mass percentage that the solvent accounts for the gumbo eye drops is 4%-96%；
The solvent is water or PBS；
The gumbo extract is made by following preparation method, the preparation method the following steps are included:
(1) gumbo pod is cleaned, chopping, homogenate, carries out water extraction, obtains crude extract；
(2) the crude extract ethanol precipitation for obtaining step (1) is centrifuged, dry, obtains jelly；
(3) jelly for obtaining step (2) dissolves, and dialyses in the distilled water of 15-20 times of volume, is freeze-dried.
2. gumbo eye drops as described in claim 1, which is characterized in that the gumbo eye drops is by gumbo extract and molten
Agent composition.
3. gumbo eye drops as described in claim 1, which is characterized in that
Gumbo pod described in step (1) is gumbo fruit, the preferably fresh pod of gumbo；
And/or the operation that water described in step (1) extracts is the following steps are included: by the distillation of gumbo pod and 5-10 times of volume
Water mixes, and stands 12-24 hours, and centrifugation removes sediment；The revolving speed of the centrifugation is 2000-4000rpm, centrifugation
Time is 2-5min；
And/or the mass percent concentration of ethyl alcohol described in step (2) be 30%-40%, preferably 35%；
And/or in step (2), the revolving speed of the centrifugation is 2000-3500rpm, and the time of centrifugation is 3-10min；
And/or in step (2), the drying is vacuum drying, and the vacuum drying temperature is 50-60 DEG C；
It is described to be dissolved as jelly being dissolved in distilled water and/or in step (3)；
And/or in step (3), the dialysis is carried out using dialysis membrane；The time of the dialysis is 24-48 hours.
4. gumbo eye drops as described in claim 1, which is characterized in that the gumbo eye drops includes following components: the autumn
Certain herbaceous plants with big flowers extract, solvent and auxiliary material；Wherein,
The mass percentage that the gumbo extract accounts for the gumbo eye drops is 4%-95%；
The mass percentage that the solvent accounts for the gumbo eye drops is 4%-95%；
The mass percentage that the auxiliary material accounts for the gumbo eye drops is 0.1%-1%；
The auxiliary material includes preservative.
5. gumbo eye drops as claimed in claim 4, which is characterized in that the preservative be selected from sodium benzoate, sorbic acid and
One of ethyl hydroxy benzoate is a variety of, preferably sodium benzoate.
6. gumbo eye drops as described in claim 4 or 5, which is characterized in that the gumbo eye drops by gumbo extract,
Solvent and auxiliary material composition.
7. gumbo eye drops as described in claim 1, which is characterized in that
The PBS is phosphate buffered saline solution, pH 7.2-7.4；
And/or the water is distilled water；
And/or the pH value of the gumbo eye drops be 4.60-4.70, preferably 4.65.
8. the gumbo eye drops as described in claim 1,2,4 or 5, which is characterized in that the gumbo extract accounts for the gumbo
The mass percentage of eye drops is 5%-95%, preferably 25% or 50%.
9. a kind of preparation method of such as described in any item gumbo eye drops of claim 1-8, which is characterized in that the preparation side
Method is the following steps are included: the gumbo extract and solvent are mixed.
10. a kind of preparation method of gumbo eye drops as claimed in claim 9, which is characterized in that if the gumbo eye drip
Liquid includes auxiliary material, then preparation method includes the following steps: by gumbo extract, solvent and the auxiliary materials and mixing.
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