CN108120782A - A kind of assay method of ivermectin chewable tablets dissolution rate - Google Patents
A kind of assay method of ivermectin chewable tablets dissolution rate Download PDFInfo
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- G—PHYSICS
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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Abstract
The present invention provides a kind of assay methods of ivermectin chewable tablets dissolution rate.Assay method of the present invention can significantly increase the ivermectin component peak area measured, so as to add the sensitivity of detection, so that the repeatability of this method, reappearance, accuracy have larger improvement, it is the dissolution determination method of really suitable ivermectin chewable tablets.Meanwhile assay method of the present invention is suitable for a variety of brands, the high performance liquid chromatograph of model and chromatographic column, improves the durability of dissolution method measure.
Description
Technical field
The present invention relates to field of chemical detection, in particular to a kind of measure side of ivermectin chewable tablets dissolution rate
Method.
Background technology
After solid pharmaceutical preparation oral medication, the absorption of drug depends on drug from the dissolution in preparation or release and in stomach and intestine
The infiltration in road.Since the dissolution and dissolving of drug have material impact to absorbing, dissolution in vitro experiment exists for drug
The prediction of internal behavior also has certain practical significance.
Based on above-mentioned consideration, oral solid formulation dissolution in vitro test method is established, there is following effect：Evaluate preparation batch
Between quality uniformity；Instruct the research and development of novel formulation；(such as prescription, production technology, Workplace after some changes occur for product
Change and production technology amplification), the uniformity of confirmation drug quality and curative effect.
When establishing dissolution rate quality standard, it should be taken into account dissolubility, permeability, dissolved corrosion and the medicine of drug for power
The factors such as learn, to ensure the uniformity of product quality before and after the uniformity of quality, change and technique amplification between drug batch.
Ivermectin chewable tablets is big ring Inner esters antiparasitic agents, for preventing dog nematosis, acariasis and parasitics elder brother
Parasitosis, main component are ivermectin.Ivermectin can be distributed to most tissues very well after absorbing, so ivermectin nozzle
Chewing the dissolution rate of active ingredient in piece becomes the pharmaceutically-active key factor of limitation ivermectin chewable tablets performance.Thus, for
The measure of ivermectin chewable tablets dissolution rate, it may have significance.
However, due to the very low only 0.8mg of the content specification of ivermectin chewable tablets, and《The Ministry of Agriculture of the People's Republic of China, MOA
Announce No. 2548》Dissolution medium is 500ml in dissolution detection method under ivermectin chewable tablets quality standard, has been
Many brand, the minimum dissolution medium amounts of model digestion instrument, so inevitably resulting in the upper machine concentration of test solution too
Low (being only 1.6 μ g/ml, close to the quantitative limit of notification number dissolution detection method), hence this method sensitivity is poor,
Larger harmful effect is caused to the stability of testing result, accuracy, repeatability.And the Ministry of Agriculture announces No. 2548 Yi Wei
The brand of high performance liquid chromatograph, model used in dissolution detection method under rhzomorph chewable tablets quality standard also have special want
Ask, found in routine testing, it is fine with 2695 detection result of Shimadzu 20A, waters, but with 1260 precision of Agilent then compared with
Difference.Therefore, if according to bulletin method to ivermectin chewable tablets carry out dissolution determination, can generate reappearance, accuracy,
The problems such as durability is poor.
In view of this, it is special to propose the present invention.
The content of the invention
The first object of the present invention is to provide a kind of assay method of ivermectin chewable tablets dissolution rate, the measure
Method has many advantages, such as that sensitive height, accuracy are high, reproducible and durability is good.
In order to realize the above-mentioned purpose of the present invention, spy uses following technical scheme：
A kind of assay method of ivermectin chewable tablets dissolution rate, the assay method include：
The ivermectin content in ivermectin chewable tablets sample is detected using high performance liquid chromatography；
Wherein, in the high performance liquid chromatography, object of reference of the ivermectin reference substance for dissolution test is used, and is adopted
With following condition：
Sample size 45~55 μ L, 238~254nm of Detection wavelength.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, the reference substance solution with
Test solution sample size is 50 μ l.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, the high performance liquid chromatography
Using following condition：
Mobile phase is that acetonitrile-methanol-water mixed solution is mobile phase, and flow velocity 1.0ml/min, column temperature is 30 DEG C, theoretical
Plate number presses H2B1aPeak meter is not less than 2000, ivermectin H2B1aWith H2B1bThe separating degree at peak is not less than 3.0.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, acetonitrile, methanol in mobile phase
And the volume ratio of water is (50~60)：(30~40)：(10~20).
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, acetonitrile, methanol in mobile phase
And the volume ratio of water is 53：35：12.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, the assay method includes：
In the high performance liquid chromatography detection, ivermectin chewable tablets is taken to be dissolved out with sodium dodecyl sulfate solution, made
For test solution；
In the high performance liquid chromatography detection, ivermectin reference substance is taken, methanol is added to dissolve and quantify, then with 12
Sodium alkyl sulfate solution dilutes, as reference substance solution.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, the dodecyl sulphate
The mass concentration of sodium solution is 0.4%, and test sample dissolution dosage is 500ml.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, ivermectin chewable tablets is molten
The rotating speed gone out is 75r/min, dissolution time 45min.
Preferably, in the assay method of ivermectin chewable tablets dissolution rate of the present invention, Yi Wei in reference substance solution
The content of rhzomorph is 2 μ g/ml.
Compared with prior art, beneficial effects of the present invention are：
1. assay method of the present invention can significantly increase the ivermectin component peak area measured, so as to add the spirit of detection
Sensitivity so that the repeatability of this method, reappearance, accuracy have larger improvement, are really to be suitble to ivermectin chewable tablets
Dissolution determination method.
2. ivermectin chewable tablets dissolution method of the present invention is suitable for a variety of brands, the high-efficient liquid phase color of model
Spectrometer and chromatographic column improve the durability of dissolution method.
Specific embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment, but those skilled in the art will
Understand, the following example is merely to illustrate the present invention, and is not construed as limiting the scope of the invention.It is not specified in embodiment specific
Condition person, the condition suggested according to normal condition or manufacturer carry out.Reagents or instruments used without specified manufacturer is
The conventional products that can be obtained by commercially available purchase.
Being analyzed etc. for drug evaluation and materia medica in view of dissolution rate has important reference significance, and existing bulletin
Standard method can not all meet the practical problems such as actual demand in precision, accuracy and repeatability, and the present invention provides one
The assay method of kind of ivermectin chewable tablets dissolution rate, by the preferred of sample size and Detection wavelength, establishing a kind of Gao Ling
The good dissolution determination method of sensitivity, high accuracy, repeatability and durability.
Specifically, the method for the present invention includes the following steps：
It is prepared by test solution：It is filtered after the dissolution of ivermectin chewable tablets；Preferably, the dissolution of ivermectin chewable tablets is
It is what dissolution medium carried out by 0.4% sodium dodecyl sulfate solution of mass concentration；
The dodecyl sulphur for being 0.4% by the ivermectin chewable tablets input 500ml mass concentrations that content specification is 0.8mg
After acid sodium solution, the disintegration of piece and the dissolution of ivermectin are carried out by chew under leaching condition；And the rotating speed of stirring and dissolving is
75r/min, time 45min；
Then appropriate dissolution fluid is taken, is filtered, it is test sample solution to take subsequent filtrate；
It is prepared by reference substance solution：Preferably, in this step, it is to add in 10mg ivermectins in 10ml volumetric flasks, adds first
Alcohol dissolves and is diluted to scale, then shakes up；Then, appropriate acquired solution is taken, then with the dodecane that mass concentration is 0.4%
Base metabisulfite solution dilutes, and the concentration of ivermectin in gained reference substance solution is caused to be diluted to 2 μ g/ml.
After prepared by test sample solution and reference substance solution, i.e., the detection of dissolution rate, tool are carried out using high performance liquid chromatography
Body step is with reference to as follows：
Reference substance solution and each 45~55 μ l of test solution, preferably 50 μ l injection high performance liquid chromatography are drawn respectively
Instrument carries out ivermectin content detection using high performance liquid chromatography, chromatogram is recorded, by external standard method with calculated by peak area Yi Wei
Rhzomorph stripping quantity；
The chromatographic condition of high performance liquid chromatography：Using octadecylsilane binding silica gel as the filler of chromatographic column, with second
Nitrile-methanol-water mixed solution be mobile phase, 238,245 or 254nm of Detection wavelength；It is furthermore preferred that Detection wavelength is
245nm；
Flow velocity is 1.0ml/min, and column temperature is 30 DEG C, and number of theoretical plate presses H2B1aPeak meter is not less than 2000, ivermectin H2B1a
With H2B1bThe separating degree at peak is not less than 3.0；
Likewise it is preferred that, the volume ratio of acetonitrile, methanol and water is (50~60) in mobile phase：(30~40)：(10~
20)；Preferably, the volume ratio of acetonitrile, methanol and water is 53 in mobile phase：35：12.
And can substantially be learnt by assay method as above, sample introduction is improved using big absorbing wavelength simultaneously in the present invention
The mode of amount is detected, so that the peak area bigger of detection, and the sensitivity of determination data of the present invention is improved, and
So that the method for the present invention have it is good can accuracy, reappearance and durability.
Embodiment 1
Chromatographic condition and system suitability：It is filler with octadecylsilane chemically bonded silica；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 245nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Take ivermectin chewable tablets to be detected, according to dissolution rate and drug release determination method (《Chinese veterinary pharmacopoeia》
0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is 75 turns per minute, according to
Method operates, and during through 45 minutes, takes solution appropriate, filters,
Subsequent filtrate is taken as test solution；
It is another to take ivermectin reference substance 10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale,
It shakes up, precision measures in right amount, the solution for containing 2 μ g in every 1ml is made of dissolution medium, as reference substance solution.
It is accurate respectively to measure test solution and each 50 μ l injections liquid chromatograph of reference substance solution, chromatogram is recorded, is pressed
External standard method is with the stripping quantity of calculated by peak area every.Limit is the 75% of labelled amount, should meet regulation.
1 dissolution determination method of embodiment is investigated：
1st, Detection wavelength is definite
Ivermectin reference substance solution is taken, using ultraviolet spectrophotometry, is scanned in the range of 190nm~600nm, is invented
People has found that ivermectin has absorption maximum at wavelength 238nm, 245nm, i.e. ivermectin detects energy ratio at two wavelength
The dissolution rate Detection wavelength (254nm) that the Ministry of Agriculture is announced under No. 2548 ivermectin chewable tablets quality standard obtains larger peak
Area improves detection sensitivity.But it by further investigating, finds to detect the dissolution of ivermectin chewable tablets at 238nm wavelength
In the collection of illustrative plates of degree, ivermectin peak and adjacent peak baseline separation effect are poor, and best separation can be obtained in 245nm wavelength detectings
Effect, therefore preferably 245nm is optimal wavelength.
2. sample size determines
Screening is optimized to different sample sizes respectively, it, will be with portion Yi Wei bacterium according to the method and condition of embodiment 1
Plain chewable tablets test solution repeats sample introduction 6 times, measures peak area respectively.
Sample size screening experiment result：
From testing result as above, sample test solution peak area under the conditions of 50 μ l sample sizes is most consistent, side
Method sensitivity is best, when sample size is less than 50 μ l, more deviates 50 μ l, and RSD is bigger, shows that sample size is smaller, between every pin
Relative standard deviation is bigger, is difficult to guarantee the accuracy of detection；And when sample size be more than 50 μ l when, RSD has change trend, this be because
It is excessive for sample size, cause overload phenomenon, peak shape substantially broadens, and theoretical pedal number and separating degree are deteriorated, and detection efficiency declines,
Therefore selected 50 μ l are optimal sample size.
3. specificity is tested
The preparation of negative control solution：Other raw materials and the auxiliary material in addition to ivermectin are weighed in prescription ratio, according to her
The preparation process and test solution for tieing up rhzomorph chewable tablets prepare method below item and negative control solution are made.
Precision draws test solution, reference substance solution, negative control solution, each 50 μ l of mobile phase, is established by the present invention
Content assaying method chromatographic condition sample introduction.
Result of the test shows that test sample chromatographic peak, and test sample chromatographic peak occurs on a corresponding position with reference substance
Middle baseline is steady, separating degree is good；Negative control, mobile phase occur on the position without chromatographic peak, show mobile phase and feminine gender
It is noiseless to impinging upon ivermectin retention time.
4. the stability of sample solution
Take test solution, point stand 0,2,4,6,10 it is small when, using identical chromatographic conditions, sample introduction 50 μ l are as a result as follows
Shown in table：
According to chromatographic peak area measured in such as upper table, test sample group RSD is 1.24% ＜ 3.0%, prompts reference substance
And test solution is good in 10h internal stabilities.
5. linear relationship
Precision weighs ivermectin reference substance 10mg and is placed in 10ml measuring bottles, and methanol is added to be allowed to be completely dissolved rear constant volume, is shaken
Even, precision measures 8ml and is placed in 250ml measuring bottles, with dissolution medium constant volume, shakes up, and the solution conduct for containing 32 μ g in every 1ml is made
Storing solution.The storing solution 3,4,5,6,7ml are measured again respectively in 100ml measuring bottles, scale is diluted to dissolution medium, shakes up,
Obtain 60%, 80%, 100%, 120%, 140% Series Measurement strength solution.By the chromatographic condition that the present invention establishes, sample introduction 50
μ l, determination sample chromatographic peak area, the results are shown in table below：
Using the peak area in as above form and concentration data as coordinate, standard curve is established, obtaining equation is：Y=
37.688X-1.3(R2=0.9995), prompt ivermectin concentrations in the range of the μ g/ml of 0.96 μ g/ml~2.24 with peak area
There are good linear relationships.
6. precision test
By as above test 5. linear relationships experiment in test concentrations be adjusted to 100% solution it is (dense i.e. containing ivermectin
Spend for the solution of 1.6 μ g/ml), under identical chromatographic conditions, repeat sample introduction 6 times, 50 μ l, measure peak area respectively every time, investigate
Instrument precision, and the relative standard deviation RSD obtained by 6 groups of data is calculated, the results are shown in table below：
From as above list data result, the standard deviation for repeating to test six times is 1.51% ＜ 3.0%, it can be seen that
The dissolution determination method precision that the present invention optimizes is good, Precision test result meet regulation (sample size 0.32%,
Between 0.1%~1%, according to《Chinese Pharmacopoeia》Version in 2015《Drug standard analysis method verification guide principle》It is accurate
RSD tolerance intervals regulation is spent, 3%) RSD is not greater than
7. repetitive test
Selection after accurately weighed, according to test sample item operation preparation solution, is used with 6 parts of lot number ivermectin chewable tablets
Identical chromatographic conditions, 50 μ l of sample introduction gather the chromatogram of 6 parts of samples, calculate the relative standard deviation RSD of solution, as a result as follows
Shown in table：
The standard deviation measured by 6 parts of samples of property results showed that repeated as above is 2.72% ＜ 3.0%,
It can be seen that the dissolution determination method repeatability that the present invention optimizes is good, repetitive test result also complies with rule
Fixed (sample size 0.32%, between 0.1%~1%, according to《Chinese Pharmacopoeia》2015
Year version《Drug standard analysis method verification guide principle》Repeated RSD tolerance intervals regulation,
3%) RSD is not greater than.
8. reappearance test
Selection is with 6 parts of lot number ivermectin chewable tablets, by another analyst on different experiments room, different working days,
Using different liquid chromatographs, different chromatographic columns is operated with " repetitive test ", measures the dissolution of ivermectin chewable tablets
Degree, the results are shown in table below：
It is 0.38% ＜ 6.0% by the RSD values of results showed that analyst's A, B analysis result as above, difference is
0.52%, meet regulation (sample size 0.32%, between 0.1%~1%, according to《Chinese Pharmacopoeia》Version in 2015《Drug
Quality standard analysis method verification guide principle》Reappearance RSD tolerance intervals provide 6%) RSD is not greater than.
It can be seen that the dissolution determination method reappearance that the present invention optimizes is good.
9. accuracy test
Ivermectin reference substance and full auxiliary material mixture are weighed in 80%, 100%, 120% prescription ratio precision, each 3 parts,
It is uniformly mixed, each 3 parts of the accuracy test print of three kinds of concentration gradients is made according to the preparation process of ivermectin chewable tablets, by this
The dissolution determination method of invention optimization measures ivermectin dissolution rate, calculates the rate of recovery, the results are shown in table below：
By results showed that as above, the rate of recovery and average recovery rate in 90%~108%, RSD within 3%,
Meet regulation (sample size 0.32%, between 0.1%~1%, according to《Chinese Pharmacopoeia》Version in 2015《Drug quality mark
Quasi- analysis method verification guide principle》Rate of recovery limit tolerance interval provides that rate of recovery limit should be 90%~108%)
It can be seen that the dissolution determination method accuracy that the present invention optimizes is good.
10. serviceability test
Take appropriate with a collection of ivermectin chewable tablets, investigate change in flow ± 0.2ml/min respectively, column temperature change ± 5 DEG C,
Methanol ratio changes ± 5%, different chromatographic columns, the efficient liquid of different model in acetonitrile ratio variation ± 5%, mobile phase in mobile phase
The variation of instrument chromatographic behavior during chromatography, observation base separation situation, calculate it is each under the conditions of the data obtained average value and
RSD values, serviceability test result are as follows：
By in results showed that ivermectin chewable tablets dissolution determination method serviceability test as above, flow velocity, column
Temperature, chromatographic column, high performance liquid chromatograph model, the small variations for flowing phase composition, ivermectin peak and adjacent peak reach base
Line separates, and the equal ＜ 3% of RSD values of ivermectin dissolution rate is surveyed under the conditions of each.It can be seen that the dissolution rate that the present invention optimizes is surveyed
Determine method good tolerance.
11. quantitative limit
Ivermectin reference substance solution is taken, is dissolved with dissolution medium, and is diluted step by step, adjusting reference substance solution concentration makes her
The response of dimension rhzomorph is about 10 times of noise level, show that quantifying for ivermectin is limited to 0.75 μ g/ml, and in reference substance she
The concentration of dimension rhzomorph is about 1.6 μ g/ml, and the concentration of ivermectin is about 2.0 μ g/ml in test sample, it can be seen that the present invention is excellent
The dissolution determination method quantitative limit of change can meet detection needs.
12. ivermectin dissolution determination in sample
The dissolution determination method optimized by the present invention measures ivermectin in three batches of ivermectin chewable tablets and dissolves out respectively
Degree, it is as a result as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 95.8 | 97.5 | 96.6 |
| 2 | 96.7 | 98.0 | 98.4 |
| 3 | 97.3 | 95.7 | 95.9 |
| Average value (%) | 96.6 | 97.1 | 97.0 |
| RSD (%) | 0.78 | 1.25 | 1.33 |
The result shows that the ivermectin dissolution rate RSD of three batches of samples is respectively less than 3%, difference is small in batch.
Embodiment 2
Chromatographic condition is filler with octadecylsilane chemically bonded silica with system suitability；With acetonitrile-methanol-water
(53：35：12) it is mobile phase；Detection wavelength 238nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin H2B1a
With H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aThe 0.8 of peak
Times.
Assay method：Take ivermectin chewable tablets sample to be measured, according to dissolution rate and drug release determination method (《Chinese veterinary drug
Allusion quotation》0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is 75 turns per minute,
It operates in accordance with the law, during through 45 minutes, takes solution appropriate, filter, take subsequent filtrate as test solution；Separately take ivermectin reference substance
10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale, is shaken up, precision measures in right amount, is situated between with dissolution
The solution for containing 2 μ g in every 1ml is made in matter, as reference substance solution.Each accurate 50 μ l that measure inject liquid chromatograph, record chromatography
Figure, by external standard method with the stripping quantity of calculated by peak area every.
Ivermectin dissolution rate in three batches of ivermectin chewable tablets is measured respectively by the dissolution determination method of embodiment 2,
As a result it is as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 92.6 | 98.5 | 97.7 |
| 2 | 97.8 | 91.7 | 99.0 |
| 3 | 96.6 | 92.6 | 93.4 |
| Average value (%) | 95.7 | 94.3 | 96.7 |
| RSD (%) | 2.85 | 3.92 | 3.03 |
As above measurement result shows that the ivermectin dissolution rate RSD of three batches of samples is unstable or even also greater than 3%, batch
Interior difference is big, this is because when detecting ivermectin chewable tablets dissolution rate at 238nm wavelength, inferior separating effect causes to detect
Result error is exceeded.
It can be seen that for compared to 238nm Detection wavelengths, the measurement result of 245nm Detection wavelengths is more accurate.
Embodiment 3
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 254nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Ivermectin chewable tablets is taken as test article, according to dissolution rate and drug release determination method (《Chinese veterinary drug
Allusion quotation》0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is 75 turns per minute,
It operates in accordance with the law, during through 45 minutes, takes solution appropriate, filter, take subsequent filtrate as test solution；Separately take ivermectin reference substance
10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale, is shaken up, precision measures in right amount, is situated between with dissolution
The solution for containing 2 μ g in every 1ml is made in matter, as reference substance solution.Each accurate 50 μ l that measure inject liquid chromatograph, record chromatography
Figure, by external standard method with the stripping quantity of calculated by peak area every.
Ivermectin dissolution rate in three batches of ivermectin chewable tablets is measured respectively by the dissolution determination method of embodiment 3,
As a result it is as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 93.6 | 91.1 | 94.4 |
| 2 | 92.8 | 92.3 | 95.9 |
| 3 | 94.6 | 93.7 | 92.8 |
| Average value (%) | 93.7 | 92.4 | 94.4 |
| RSD (%) | 0.96 | 1.41 | 1.64 |
The result shows that the ivermectin dissolution rate RSD of three batches of samples is respectively less than 3%, difference is small in batch, but compared with embodiment 1
RSD values it is big, this is because 245nm is the maximum absorption wavelength of ivermectin, thus the peak area detected at 254nm wavelength compared with
245nm is small, causes method sensitivity poor, it is larger to measure result error.
Comparative example 1
According to《The Ministry of Agriculture of the People's Republic of China, MOA announces No. 2548》Middle method is to the dissolution rate of ivermectin chewable tablets
It is detected, actual conditions is as follows：
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 254nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Ivermectin chewable tablets is taken as sample to be tested, according to dissolution rate and drug release determination method (《Chinese beast
Pharmacopeia》0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is per minute 75
Turn, operate in accordance with the law, during through 45 minutes, take solution appropriate, filter, take subsequent filtrate as test solution；Separately take ivermectin pair
It is accurately weighed according to product 10mg, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale, is shaken up, precision measures in right amount, and use is molten
Go out medium and the solution for containing 2 μ g in every 1ml is made, as reference substance solution.Each accurate 20 μ l that measure inject liquid chromatograph, record
Chromatogram, by external standard method with the stripping quantity of calculated by peak area every.
Ivermectin dissolution rate in three batches of ivermectin chewable tablets is measured respectively by the dissolution determination method of comparative example 1,
As a result it is as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 85.6 | 73.3 | 79.7 |
| 2 | 92.8 | 95.5 | 88.4 |
| 3 | 97.7 | 80.3 | 95.6 |
| Average value (%) | 92.0 | 83.0 | 87.9 |
| RSD (%) | 6.61 | 13.67 | 9.06 |
It from testing result as above, is measured according to the method for comparative example 1, the ivermectin dissolution of three batches of samples
Degree RSD is all higher than 3%, widely different in batch, and it is very big to measure result error, this is because with this method and high performance chromatograph model
When detecting ivermectin chewable tablets dissolution rate, the upper machine concentration of the method is close to caused by quantitative limit so that measures peak area mistake
Small, precision is very poor.
Comparative example 2
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 245nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Ivermectin chewable tablets is taken as detected sample, according to dissolution rate and drug release determination method (《China
Veterinary drug allusion quotation》0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is per minute
It 75 turns, operates in accordance with the law, during through 45 minutes, takes solution appropriate, filter, take subsequent filtrate as test solution；Separately take ivermectin
Reference substance 10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale, is shaken up, precision measures appropriate, uses
The solution for containing 2 μ g in every 1ml is made in dissolution medium, as reference substance solution.Each accurate 30 μ l that measure inject liquid chromatograph, note
Chromatogram is recorded, by external standard method with the stripping quantity of calculated by peak area every.
Ivermectin dissolution rate in three batches of ivermectin chewable tablets is measured respectively by the dissolution determination method of comparative example 2,
As a result it is as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 87.4 | 96.2 | 86.3 |
| 2 | 95.6 | 92.8 | 90.1 |
| 3 | 92.6 | 89.7 | 93.5 |
| Average value (%) | 91.9 | 92.9 | 90.0 |
| RSD (%) | 4.52 | 3.50 | 4.00 |
It from testing result as above, is measured according to the method for comparative example 2, the ivermectin dissolution of three batches of samples
Degree RSD is all higher than 3%, and difference is big in batch, hence it is evident that it is larger to measure result error than embodiment 1.This is because 30 μ l sample sizes compared with
It is small, cause to measure that peak area is too small, and method sensitivity is poor, and it is also larger to measure result error.
Comparative example 3
Chromatographic condition and system suitability：Using octadecylsilane chemically bonded silica as filler；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 245nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Ivermectin chewable tablets is taken as detected sample, according to dissolution rate and drug release determination method (《China
Veterinary drug allusion quotation》0,931 second method of annex), using 0.4% sodium dodecyl sulfate solution 500ml as dissolution medium, rotating speed is per minute
It 75 turns, operates in accordance with the law, during through 45 minutes, takes solution appropriate, filter, take subsequent filtrate as test solution；Separately take ivermectin
Reference substance 10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale, is shaken up, precision measures appropriate, uses
The solution for containing 2 μ g in every 1ml is made in dissolution medium, as reference substance solution.Each accurate 70 μ l that measure inject liquid chromatograph, note
Chromatogram is recorded, by external standard method with the stripping quantity of calculated by peak area every.
Ivermectin dissolution rate in three batches of ivermectin chewable tablets is measured respectively by the dissolution determination method of comparative example 2,
As a result it is as follows：
| Piece number | 20170801 | 20170802 | 20170803 |
| 1 | 98.4 | 96.4 | 92.9 |
| 2 | 99.2 | 90.7 | 99.6 |
| 3 | 91.5 | 98.6 | 90.1 |
| Average value (%) | 96.4 | 95.2 | 94.2 |
| RSD (%) | 4.39 | 4.28 | 5.18 |
It from testing result as above, is measured according to the method for comparative example 3, the ivermectin dissolution of three batches of samples
Degree RSD is all higher than 3%, and difference is big in batch, hence it is evident that it is larger to measure result error than embodiment 1.This is because 70 μ l sample sizes compared with
Greatly, cause overload phenomenon occur, peak shape broadens, and detection efficiency declines, and separating effect is poor, and dissolution rate testing result is than true
Value has suspicion bigger than normal.
To sum up the measurement result of embodiment 1-5 and comparative example 1-3 understand, ivermectin chewable tablets dissolution determination it is excellent
The condition is selected to be：
Chromatographic condition and system suitability：It is filler with octadecylsilane chemically bonded silica；With acetonitrile-methanol-
Water (53：35：12) it is mobile phase；Detection wavelength 245nm.Number of theoretical plate presses H2B1aPeak is calculated not less than 2000, ivermectin
H2B1aWith H2B1bThe separating degree at peak should be not less than 3.0, ivermectin H2B1bThe retention time at peak is about ivermectin H2B1aPeak
0.8 times.
Assay method：Ivermectin chewable tablets to be detected is taken, using 0.4% sodium dodecyl sulfate solution 500ml as dissolution
Medium, rotating speed are 75 turns per minute, are operated in accordance with the law, during through 45 minutes, take solution appropriate, filter, take subsequent filtrate as test sample
Solution；
It is another to take ivermectin reference substance 10mg, it is accurately weighed, it puts in 10ml measuring bottles, methanol is added to dissolve and is diluted to scale,
It shakes up, precision measures in right amount, the solution for containing 2 μ g in every 1ml is made of dissolution medium, as reference substance solution.
It is accurate respectively to measure test solution and each 45~55 μ l injections liquid chromatograph of reference substance solution, record chromatography
Figure, by external standard method with the stripping quantity of calculated by peak area every.
And according to assay method as above can not only Accurate Determining ivermectin chewable tablets dissolution rate, and with good
Good sensitivity and repeatability, and with good durability and reappearance.
Although illustrate and describing the present invention with specific embodiment, it will be appreciated that without departing substantially from the present invention's
Many other change and modification can be made in the case of spirit and scope.It is, therefore, intended that in the following claims
Including belonging to all such changes and modifications in the scope of the invention.
Claims (9)
1. a kind of assay method of ivermectin chewable tablets dissolution rate, which is characterized in that the assay method includes：
The ivermectin content in ivermectin chewable tablets sample is detected using high performance liquid chromatography；
Wherein, in the high performance liquid chromatography, object of reference of the ivermectin reference substance for dissolution test is used, and using such as
Lower condition：
Sample size 45~55 μ L, 238~254nm of Detection wavelength.
2. the assay method of ivermectin chewable tablets dissolution rate according to claim 1, which is characterized in that the reference substance
Solution and test solution sample size are 50 μ l.
3. the assay method of ivermectin chewable tablets dissolution rate according to claim 1, which is characterized in that the efficient liquid
Phase chromatography uses following condition：
Mobile phase is that acetonitrile-methanol-water mixed solution is mobile phase, and flow velocity 1.0ml/min, column temperature is 30 DEG C, number of theoretical plate
By H2B1aPeak meter is not less than 2000, ivermectin H2B1aWith H2B1bThe separating degree at peak is not less than 3.0.
4. the assay method of ivermectin chewable tablets dissolution rate according to claim 3, which is characterized in that second in mobile phase
The volume ratio of nitrile, methanol and water is (50~60)：(30~40)：(10~20).
5. the assay method of ivermectin chewable tablets dissolution rate according to claim 4, which is characterized in that second in mobile phase
The volume ratio of nitrile, methanol and water is 53：35：12.
6. the assay method of ivermectin chewable tablets dissolution rate according to claim 1, which is characterized in that the measure side
Method includes：
In the high performance liquid chromatography detection, ivermectin chewable tablets is taken to be dissolved out with sodium dodecyl sulfate solution, as confession
Test sample solution；
In the high performance liquid chromatography detection, ivermectin reference substance is taken, methanol is added to dissolve and quantify, then with dodecyl
Metabisulfite solution dilutes, as reference substance solution.
7. the assay method of ivermectin chewable tablets dissolution rate according to claim 6, which is characterized in that the dodecane
The mass concentration of base metabisulfite solution is 0.4%, and test sample dissolution dosage is 500ml.
8. the assay method of ivermectin chewable tablets dissolution rate according to claim 6, which is characterized in that ivermectin nozzle
The rotating speed for chewing piece dissolution is 75r/min, dissolution time 45min.
9. the assay method of ivermectin chewable tablets dissolution rate according to claim 6, which is characterized in that reference substance solution
The content of middle ivermectin is 2 μ g/ml.
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