CN109432012A - A kind of preparation method of baicalein medicament-carried nano stick - Google Patents
A kind of preparation method of baicalein medicament-carried nano stick Download PDFInfo
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Abstract
The present invention relates to a kind of preparation methods of baicalein medicament-carried nano stick, belong to pharmaceutical field, it the described method comprises the following steps: weighing baicalein respectively first and pectin is added in glass container, add phosphate buffer solution, glass container is immersed in 50 DEG C of waters bath with thermostatic control, magnetic agitation mixes, and obtains mixed liquor I；Ca (OH) aqueous solution is added dropwise into mixed liquor I again, reheats 1h under the conditions of 50 DEG C；Then NaHCO aqueous solution is added dropwise again, reheats 3h under the conditions of 50 DEG C, obtains mixed liquor II；Baicalein finally is added into mixed liquor II, continuing magnetic force stirring for 24 hours, is fitted into bag filter after cooling, is dialysed for 24 hours in 20 DEG C of phosphate buffer solution, obtain baicalein medicament-carried nano stick under the conditions of 50 DEG C.Preferable targeting, histocompatbility, less toxic side effect and slow-release function are shown using baicalein medicament-carried nano stick prepared by the preparation method.
Description
Technical field
The invention belongs to pharmaceutical fields, and in particular, to a kind of preparation method of baicalein medicament-carried nano stick.
Background technique
Nanometer medicine-carried system refers to prepared nanostructure partial size between 1~1000nm, drug is encapsulated, dispersion,
It is adsorbed in polymer and is formed by integral framework.Flavone compound refers to that with chromone ring and phenyl ring be the one of basic structure
The general name of class compound is the derivative of chromone or chromogen alkane, with flavones (2- phenyl chromone) be parent nucleus and derivative one
Class yellow pigment.Nano medicament carrying system is a kind of novel form of administration, and the size and body protein of drug-carrying nanometer particle are similar,
Easily enter in tissue fluid through cell membrane without being repelled by body, there is good histocompatbility.By drug encapsulation
Get up to realize slow releasing function and stablize the effect of drug, reduces drug localized clusters degree, it is possible to reduce to the bad of body
Reaction.By the way that the carrier material for having special compatibility to body different tissues is surface modified or is encapsulated in drug
In, target-oriented drug can be improved.And carrier material is easy to get, and is essentially all natural and synthesis high polymer, and
And preparation process mild condition, method is terse, and not needing special precision instrument and equipment can be made into.Both at home and abroad for nano drug-carrying
The research of system has very big effect, and process route is reliable and stable, method is consummate, quality is stable, is suitable for industrialized production and life
Demand living, has direct drug therapy potentiality.
Baicalein (baicalein) is flavone compound, is one of the principle active component of Lamiaceae plant radix scutellariae.Tool
Have antibacterial, it is antiviral, inhibit inflammatory reaction, liver protection, cholagogue, diuresis, anticancer the effects of, have good clinical value.
Baicalein oral toxicity is extremely low, but its intravenous injection is more much bigger than oral administration gavage toxicity.In view of baicalein intravenous injection toxicity,
It is prepared into that carrier system can have targeting, good histocompatbility, less toxic side effect and to have slow-release function etc. excellent
Point.How to prepare nanoscale pharmaceutical system and then needs stringent preparation process.
Summary of the invention
For above situation, the purpose of the present invention is to provide a kind of preparation methods of baicalein medicament-carried nano stick, use
The baicalein medicament-carried nano stick of preparation method preparation show preferable targeting, histocompatbility, less toxic side effect and
Slow-release function.
To achieve the goals above, the present invention use the specific scheme is that
A kind of preparation method of baicalein medicament-carried nano stick, comprising the following steps:
Step 1: weighing 0.01g baicalein respectively and 0.1g pectin is added in glass container, phosphate buffer solution is added
Glass container is immersed in 50 DEG C of waters bath with thermostatic control by 20mL, and magnetic agitation 0.5h is mixed, and obtains mixed liquor I；
Step 2: the Ca (OH) that 8mL concentration is 0.02mol/L is added dropwise into mixed liquor I2Aqueous solution, under the conditions of 50 DEG C
Reheat 1h；Then the NaHCO that 8mL concentration is 0.03mol/L is added dropwise again3Aqueous solution reheats 3h under the conditions of 50 DEG C,
Obtain mixed liquor II；
Step 3: 0.01g baicalein is added into mixed liquor II, continuing magnetic force stirring for 24 hours, is subsequently placed in 20 under the conditions of 50 DEG C
DEG C room temperature water-bath in cooling 0.5 hour, obtain reaction mixture；The reaction mixture is fitted into bag filter, at 20 DEG C
It dialyses in the phosphate buffer solution of 500mL for 24 hours, obtains baicalein medicament-carried nano stick.
It is advanced optimized as to above scheme, the pH of the phosphate buffer solution is 6.7.
Advanced optimized as to above scheme, the phosphate buffer solution the preparation method comprises the following steps: weighing respectively
7.957g sodium dihydrogen phosphate and 17.549g disodium hydrogen phosphate powder, it is dissolved in distilled water respectively, then moves to 500ml capacity
In bottle, 500mL is settled to get phosphate buffer solution with distilled water dilution.
The utility model has the advantages that
The release that drug can effectively be delayed using baicalein medicament-carried nano stick prepared by the method for the present invention, is had certain
Slow releasing function, additionally it is possible to reduce toxic side effect, and improve the bioavilability of drug.The encapsulating of baicalein rod-like nano medicine-carried system
Rate is high, and the encapsulation rate of pectin baicalein nanometer rods reaches as high as 84.17%.And either encapsulation rate or drugloading rate are all relatively steady
It is fixed.Tablets in vitro test proves that the release of drug reaches 93.0%, and the drug accumulation release rate after dialysis for 24 hours is
67.54%.In addition, the adhesiveness of nano medicament carrying system is very big and partial size very little, anelasticity when not only improving local application
Increase, is also beneficial to increase time of contact and the contact area of drug and gastrointestinal wall, improves the biological utilisation of Oral drug absorption
Degree, therefore medicament-carried nano system can be used as the novel form of slow releasing pharmaceutical, is able to extend circulation time in vivo, works well.
The method of the invention provides theoretical basis to illustrate the dosage form research of certain pathogenesis, new drug, and baicalein will have wider
Wealthy application prospect.
Detailed description of the invention
Fig. 1 is pectin baicalein Electronic Speculum transmission plot；
Specific embodiment
Below in conjunction with the embodiment of the present invention, technical scheme in the embodiment of the invention is clearly and completely described.
Embodiment 1
1, testing program
A kind of preparation method of baicalein medicament-carried nano stick, comprising the following steps:
(1) 7.957g sodium dihydrogen phosphate and 17.549g disodium hydrogen phosphate powder are weighed respectively, it is dissolved in appropriate steaming respectively
It in distilled water, then moves in 500ml volumetric flask, it is molten for 6.7 phosphoric acid buffer to get pH value to be settled to 500mL with distilled water dilution
Liquid；
(2) accurately weighing 0.01g baicalein is that 0.10g pectin is added in 50mL volumetric flask, addition magneton, then plus
Enter the PBS solution 20mL that pH is 6.7, be then submerged in 50 DEG C of waters bath with thermostatic control, carry out magnetic agitation, heats 0.5 hour；It prepares
Ca(OH)28mL is added dropwise into above-mentioned solution in solution (0.02mol/L), is stirred at 50 DEG C 1 hour；Then, will
NaHCO38mL is added dropwise into above-mentioned reaction system in solution (0.03mol/L) dropwise, stirs other 3 hours at 50 DEG C, then plus
Enter 0.010g baicalein.The mixed system carries out continuing stirring 24 hours at 50 DEG C, then fast in 20 DEG C of room temperature water-bath
Quickly cooling but 0.5 hour.Finally, reaction mixture is fitted into bag filter, dialyse 24 hours in 20 DEG C of 500mLPBS solution,
Medicament-carried nano stick can be obtained.
2, the measurement of encapsulation rate
An important factor for encapsulation rate is evaluation preparation process thereof and quality good or not.Encapsulation rate, which refers to, to be contained in liposome
The ratio of the total dosage of dose Zhan, specific assay method is as follows for encapsulation rate in this experiment: precision measures medicament-carried nano stick sample
Solution 1mL is placed in the interior liquid of work in bag filter using the PBS buffer solution of pH 7.4 as external solution and changes a water every 12h, dialyses
24h.After dialysis, take out in liquid, add 9mL demulsifier (ethyl alcohol: ether=7: 2) be demulsified after, measure the absorbance of curcumin, will
The sample absorbance measured substitutes into absorbance-concentration relationship equation:
In A=0.000174354+0.03087c,
The concentration that medicament-carried nano stick can be measured, further according to encapsulation rate calculation formula:
The medication amount encapsulated in encapsulation rate (EE%)=system/dispensing total amount × 100%, then computational envelope rate such as the following table 1
It is shown.
Table 1: the encapsulation rate of sample
Measurement result: encapsulation rate is up to 84.17%.
3, the measurement of drugloading rate
Three groups of tests are carried out simultaneously, take 3mL sample to be placed in culture dish with microscale sampler, by sample in freeze drier
Product freeze-drying, the time 2-3 days, takes out sample after freeze-drying, scrapes freeze-dried powder, weighs in electronic balance to get medicament-carried nano stick
Total weight, drugloading rate is calculated by following formula:
Content/microballoon of drug or total weight × 100% of vesica in drugloading rate (DL%)=microballoon or vesica
Measurement result is as shown in table 2.
Table 2: the drugloading rate of sample
| Drugloading rate/% | Drugloading rate/% | Drugloading rate/% | Average value/% | |
| Sample | 18.74 | 16.88 | 18.54 | 18.05±5.65 |
Measurement result: drugloading rate is up to 18.74%.
4, the Drug Releasing Test of baicalein
1. baicalein sterling is dialysed:
Baicalein sterling 0.100g is weighed with electronic balance precision, is dissolved in the PBS solution that pH value is 6.7
In the volumetric flask of 100mL, it is configured to the baicalein titer of 1mg/mL.It takes 2.0mL to be placed in bag filter with pipette, then will
It releases the drug in the PBS solution that bag filter comprising titer is 6.7 as the pH value of 100mL, while 37 DEG C of constant temperature gas bath oscillators
Middle shaking.
Then the primary absorbance for measuring drug in the outer PBS solution of bag filter of sampling at regular intervals, records and calculates
The baicalein cumulative release amount of each period.
2. the Drug Releasing Test of baicalein nanometer rods:
Using pectin as the nanometer rods of carrier: being configured to the nano drug-carrying solution of 1mg/mL, while carrying out three groups of tests, with shifting
Liquid pipe removes 2.0mL and is placed in bag filter, in the PBS solution for being 6.7 as the pH value of 100mL by the bag filter sealed, simultaneously
It is shaken in 37 DEG C of gas bath constant temperature oscillators, then the outer sample absorbance of the primary measurement bag filter of sampling at regular intervals, together
When new PBS solution is injected into wide-mouth bottle, record and calculate the cumulative release amount of each time, and draw baicalein nanometer rods
Cumulative release amount curve, records and calculates the release of drug.As a result as shown in table 3 below.
Table 3: pectin carrier baicalein Drug Releasing Test result table
| Number | Time interval/h | Absorbance/A | Concentration/μ g/mL | Preparation/% |
| 1 | 0 | 0 | 0 | 0 |
| 2 | 1 | 0.109 | 3.525 | 17.625 |
| 3 | 2 | 0.184 | 5.952 | 29.76 |
| 4 | 4 | 0.262 | 8.476 | 42.38 |
| 5 | 6 | 0.304 | 9.845 | 49.225 |
| 6 | 8 | 0.335 | 10.854 | 54.27 |
| 7 | 12 | 0.354 | 11.478 | 57.39 |
| 8 | 16 | 0.370 | 11.986 | 59.93 |
| 9 | 20 | 0.401 | 12.974 | 64.87 |
| 10 | 24 | 0.417 | 13.508 | 67.54 |
Measurement result: after baicalein standard solution discharges 2 hours in PBS buffer solution, drug penetrates bag filter
1.86mg, release reach 93.0%.Pectin carrier baicalein nanometer rods dialysis for 24 hours after drug accumulation release rate be
67.54%.
5, stability test
Sample 1 and 2 after dilution is placed under the conditions of room temperature is protected from light and is saved two weeks, has seen whether precipitating or flocculate
It generates, and detects absorbance under ultraviolet-uisible spectrophotometer, and calculate whether its encapsulation rate changes, as a result such as table 4
Shown
Table 4: baicalein stability test result table
| Whether there is or not flocculent deposits | Encapsulation rate/% before placing | Encapsulation rate/% after two weeks | |
| Sample 1 | Nothing | 83.44 | 77.26 |
| Sample 2 | Nothing | 76.19 | 71.53 |
Measurement result: the nanometer rods of preparation generate after room temperature storage without flocculent deposit, and encapsulation rate reduces 5%
Left and right.It follows that drug has apparent leakage, especially its leakage can be more obvious when the temperature increases.If will preparation
Nanosuspension be added freeze drying protectant, after freeze-drying, place 1 month, almost without leakage phenomenon, stability is preferable.
6, high resolution TEM detects
The nano drug-carrying solution example of preparation is placed in transmission electron microscope and is observed, particle size and form are observed,
As a result it is illustrated in fig. 1 shown below.
Fig. 1 is using pectin as the transmission electron microscope picture of the nano medicament carrying system of carrier, it can be seen that single pectin carrier nanometer
Stick construction profile is spindle, and partial size has stronger affinity between 100-300nm, to hydrone, to construct not similar shape
State medicine-carried system provides the foundation and may.
The above content is interpreted as illustrative, to be not intended to limit the present invention protection scope.Protection scope of the present invention with
It is to those skilled in the art, right under the premise of without departing substantially from spirit and scope of the present invention subject to the content of claims
Some nonessential modifications and adaptations that the present invention makes still fall within protection scope of the present invention.
Claims (3)
1. a kind of preparation method of baicalein medicament-carried nano stick, it is characterised in that: the following steps are included:
Step 1: weighing 0.01g baicalein respectively and 0.1g pectin is added in glass container, phosphate buffer solution is added
Glass container is immersed in 50 DEG C of waters bath with thermostatic control by 20 mL, and magnetic agitation 0.5h is mixed, and obtains mixed liquor I；
Step 2: the Ca (OH) that 8 mL concentration are 0.02 mol/L is added dropwise into mixed liquor I2Aqueous solution, in 50 DEG C of conditions
Lower reheating 1h；Then the NaHCO that 8 mL concentration are 0.03 mol/L is added dropwise again3Aqueous solution adds again under the conditions of 50 DEG C
Hot 3h obtains mixed liquor II；
Step 3: 0.01g baicalein is added into mixed liquor II, continuing magnetic force stirring for 24 hours, is subsequently placed under the conditions of 50 DEG C
It is 0.5 hour cooling in 20 DEG C of room temperature water-bath, obtain reaction mixture；The reaction mixture is fitted into bag filter, at 20 DEG C
500 mL phosphate buffer solution in dialyse for 24 hours, obtain baicalein medicament-carried nano stick.
2. a kind of preparation method of baicalein medicament-carried nano stick as described in claim 1, it is characterised in that: the phosphoric acid buffer
The pH of solution is 6.7.
3. a kind of preparation method of baicalein medicament-carried nano stick as claimed in claim 1 or 2, it is characterised in that: the phosphoric acid
Buffer solution the preparation method comprises the following steps: 7.957 g sodium dihydrogen phosphates and 17.549 g disodium hydrogen phosphate powder are weighed respectively, by its point
It is not dissolved in distilled water, then moves in 500 ml volumetric flasks, it is molten to get phosphoric acid buffer to be settled to 500 mL with distilled water dilution
Liquid.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111638188A (en) * | 2020-05-27 | 2020-09-08 | 东北农业大学 | Method for measuring entrapment rate of lipid liposome |
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| CN1744827A (en) * | 2003-01-31 | 2006-03-08 | 帝斯曼知识产权资产管理有限公司 | The composition that comprises the novelty of carotenoid |
| CN104173292A (en) * | 2014-09-09 | 2014-12-03 | 重庆医科大学 | Novel insoluble drug-loading calcium pectate nanoparticle and preparation method thereof |
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| CN1744827A (en) * | 2003-01-31 | 2006-03-08 | 帝斯曼知识产权资产管理有限公司 | The composition that comprises the novelty of carotenoid |
| CN104173292A (en) * | 2014-09-09 | 2014-12-03 | 重庆医科大学 | Novel insoluble drug-loading calcium pectate nanoparticle and preparation method thereof |
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| CN111638188A (en) * | 2020-05-27 | 2020-09-08 | 东北农业大学 | Method for measuring entrapment rate of lipid liposome |
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