CN111721845B - Synchronous detection method for five related substances of compound ipratropium bromide solution for inhalation - Google Patents
Synchronous detection method for five related substances of compound ipratropium bromide solution for inhalation Download PDFInfo
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- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
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Abstract
The invention relates to a synchronous detection method of five related substances of a compound ipratropium bromide solution for inhalation, belonging to the technical field of drug quality determination methods. The detection method can effectively separate the ipratropium bromide, the salbutamol sulfate and other impurities, and has the characteristics of simple operation, comprehensive, accurate and reliable result and strong specificity.
Description
Technical Field
The invention relates to the technical field of medicine quality measurement methods, in particular to a synchronous detection method for five related substances of a compound ipratropium bromide solution for inhalation.
Background
The compound ipratropium bromide solution for inhalation is colorless or almost colorless clear liquid, and comprises ipratropium bromide and salbutamol sulfate as main components. The product is suitable for patients in need of multiple bronchodilators for use in combination, and is used for treating reversible bronchospasm associated with airway obstruction diseases. Wherein, ipratropium bromide and salbutamol are added to act on the muscarinic and beta 2 adrenergic receptors of the lung to produce bronchodilatory effect, and the curative effect is better than that of single administration.
The use of phosphoric acid to adjust pH to 3.2 in the case of chromatography columns of phenomenomex-C8 (4.6mm.times.250 mm,5 μm) and of ultimate XB-C8 (4.6mm.times.250 mm,5 μm), mobile phase A being acetonitrile-0.25% sodium heptanesulfonate solution (29:71), mobile phase B being acetonitrile, is disclosed in "reversed-phase high performance liquid chromatography for determination of related substances in ipratropium bromide solution for inhalation [ J ]. Chongqing medicine, 2007 (11): 1067-1068"; the detection wavelength is 210nm; the flow rate is 1.0ml/min; the 20. Mu.l sample volume was used to detect the relevant substances in the ipratropium bromide solution, but salbutamol sulfate and its related impurities could not be detected.
Literature HPLC method for determining content of two components of compound ipratropium bromide inhalation solution [ J ]]The method establishes a high performance liquid chromatography to synchronously measure the contents of the two components of ipratropium bromide and salbutamol sulfate in the compound ipratropium bromide inhalation solution according to the drug analysis and 2011 (5): 1067-1068 ". The method uses Inertsil ODS-3C18 (150 mm. Times.4.6 mm,5 μm) chromatographic column with mobile phase of 0.01mol. L -1 Potassium dihydrogen phosphate solution (pH 2.3 adjusted with phosphoric acid) (A) -acetonitrile (B), gradient elution, flow rate 1.0mL. Min -1 The detection wavelength is 220nm (detection of ipratropium bromide) and 276nm (detection of sand)Butylamine alcohol), column temperature 40 ℃. Results: the sample injection amounts of the ipratropium bromide and the salbutamol are respectively 0.01-0.2 mg.mL -1 And 0.05 to 1.0mg.mL -1 The linear relationship was good in the range (r was 0.9999, n=5) and the recovery (n=9) was 100.2% and 100.3%, respectively. Although the method detects the content of two components of the compound sartanol, the content of related substances in the compound sartanol is not further detected, the components of the related substances are complex, the related substances and the main components cannot be obviously separated by adopting the method, and the sensitivity and the specificity of the measurement cannot be ensured.
Because the detection standard of the related substances of the compound ipratropium bromide solution for inhalation has the defects, the research of a detection method which has strong specificity and high sensitivity and can detect the related substances of the compound ipratropium bromide solution for inhalation at one time is very important.
Disclosure of Invention
In view of the defects existing in the prior art, the invention aims to provide a synchronous detection method for five related substances of a compound ipratropium bromide solution for inhalation, which can be used for quality control in the production of the compound ipratropium bromide solution for inhalation by improving and optimizing the gradient, mobile phase, column temperature, flow rate and other aspects of gradient elution. The preparation operation of the mobile phase in the method is simple, the analysis method is stable, the sensitivity is high, the peak type is good, and various related substances can be detected at one time.
In order to achieve the object of the present invention, the inventors have finally found out, through a great deal of experiments, the following technical solutions by performing inventive work:
the synchronous detection method of five related substances of the compound ipratropium bromide solution for inhalation comprises the following steps: ipratropium bromide impurity C (2-phenyl-3-hydroxypropionic acid), salbutamol impurity B ((1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1- (4-hydroxyphenyl) ethanol), salbutamol impurity C (3-deoxysalbutamol), salbutamol impurity D (5- [ (1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1-hydroxyethyl ] -2-hydroxyphenyl), salbutamol impurity F (3, 3' - [ oxybis (methylene) ] bis [ alpha- [ [ (1, 1-dimethylethyl) amino ] methyl ] -4-hydroxybenzyl alcohol), and the detection method is high performance liquid chromatography, and gradient elution is adopted.
The preferred chromatographic conditions for high performance liquid chromatography are:
the filler of the chromatographic column is alkyl bonding silica gel;
the detection wavelength is 205-300nm;
the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is sodium heptanesulfonate phosphate buffer solution, and the mobile phase B is an organic phase;
setting the flow rate of the mobile phase to be 0.9-1.1 ml/min, and controlling the column temperature to be 30-40 ℃;
preferably, the gradient elution in the method may be set as follows:
more preferred gradient elution may be set to:
。
preferably the alkyl-bonded silica gel of the filler is octadecyl silane-bonded silica gel or octyl silane-bonded silica gel.
Preferably the organic phase in the mobile phase is methanol or acetonitrile; more preferably acetonitrile.
Preferably, the mobile phase is sodium heptanesulfonate phosphate buffer-acetonitrile, more preferably, the sodium heptanesulfonate phosphate buffer is 0.006mol/L.
The pH value of the sodium heptanesulfonate phosphate buffer is 2.0-3.5, and more preferably, the pH value of the sodium heptanesulfonate phosphate buffer is 2.5.
Preferably, the mobile phase flow rate in the process of the present invention is set to 1.0ml/min.
It should be noted that, in the specific embodiments, the method of the present invention is described in detail, and the method of the present invention is verified, which indicates that, compared with the prior art, the detection method of the present invention has the following unexpected effects:
(1) The method can effectively separate all five impurities of the compound ipratropium bromide solution for inhalation, and the purity of the main peak completely meets the requirements specified in pharmacopoeia.
(2) The method is simple and quick, can separate various related substances in the compound ipratropium bromide solution for inhalation under the same high performance liquid chromatography condition, avoids frequent liquid phase replacement in the detection process, greatly improves the working efficiency and reduces the cost.
(3) The invention has high separation degree, good repeatability and durability, short analysis time, simple operation and stable and reliable result.
Therefore, the invention can be used for quality control of the compound ipratropium bromide solution for inhalation, and provides effective guarantee for the quality of the final finished product.
Drawings
FIG. 1 HPLC (high Performance liquid chromatography) chart of compound ipratropium bromide solution mixed impurity reference substance for inhalation
FIG. 2 HPLC (high Performance liquid chromatography) chart of related substances of compound ipratropium bromide solution for inhalation
FIG. 3 thermal destructive HPLC profile of Compound ipratropium bromide solution for inhalation
FIG. 4 photodisruption HPLC spectrum of Compound ipratropium bromide solution for inhalation
FIG. 5 HPLC (high Performance liquid chromatography) spectrum of compound ipratropium bromide solution blank auxiliary material for inhalation
FIG. 6 HPLC (high Performance liquid chromatography) chart of related substances of compound ipratropium bromide solution for inhalation
Detailed Description
The following are specific examples of the present invention, and the technical solutions of the present invention are further described, but the scope of the present invention is not limited to these examples. All changes and equivalents that do not depart from the gist of the invention are intended to be within the scope of the invention.
Example 1
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is regulated to 2.5 to be a mobile phase A, acetonitrile is used as a mobile phase B, the detection wavelength is 210nm, the column temperature is 35 ℃, and the flow rate is 1.0ml/min.
The gradient elution is set as follows:
the test steps are as follows:
wherein ipratropium bromide and related impurity controls are purchased from Zhejiang Yougang pharmaceutical chemical company, inc.; salbutamol sulphate and related impurity controls were purchased from beijing wavile chemical company limited; the ipratropium bromide solution for inhalation is produced by Shandong New era pharmaceutical Co., ltd (lot number 06418003).
The preparation method comprises the steps of taking proper amounts of ipratropium bromide, salbutamol sulfate impurity C (2-phenyl-3-hydroxypropionic acid), salbutamol impurity B ((1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1- (4-hydroxyphenyl) ethanol), salbutamol impurity C (3-deoxysalbutamol), salbutamol impurity D (5- [ (1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1-hydroxyethyl ] -2-hydroxyphenyl) and salbutamol impurity F (3, 3' - [ oxybis (methylene) ] bis [ alpha- [ [ (1, 1-dimethylethyl) amino ] methyl ] -4-hydroxybenzyl alcohol), preparing a solution of 10 mug/ml by using 0.006mol/L sodium heptanesulfonate phosphoric acid-acetonitrile buffer solution, precisely measuring 25 mug, injecting the solution into a liquid chromatograph, and recording a chromatogram, wherein the HPLC diagram of the mixed solution is shown in the figure 1.
And injecting 25 mu l of the mobile phase mixed solution into a liquid chromatograph to obtain a spectrogram with a response value of 0-55min always about 0mAU, wherein the obtained liquidus line is basically coincident with a base line, which indicates that the detection of the relative substances of the flow has no influence.
From the spectra of fig. 1 with ipratropium bromide and salbutamol sulfate: the retention time of the main peak of ipratropium bromide is about 32min, the retention time of the main peak of salbutamol sulfate is about 14min, and the separation degree of each impurity is good.
The following table 1 of peak-out times and corresponding substances can be obtained according to fig. 1 corresponding to the localization peaks of the respective impurities.
TABLE 1 response time and corresponding substance list
| Time (min) | Corresponding substances | Relative retention time |
| 1.165 | Bromide ion | 0.04 |
| 10.195 | Ipratropium bromide C | 0.31 |
| 13.820 | Salbutamol sulfate | 0.42 |
| 17.301 | Salbutamol impurity B | 0.53 |
| 23.834 | Salbutamol impurity C | 0.73 |
| 27.240 | Salbutamol impurity D | 0.84 |
| 32.306 | Isopropiontolium bromide | 1.00 |
| 39.003 | Salbutamol impurity F | 1.20 |
2. Determination of substances related to the compound ipratropium bromide solution for inhalation: a proper amount of compound ipratropium bromide solution for inhalation is taken, and a solution containing 0.2mg/ml of ipratropium bromide is prepared by using a mixed solution of sodium heptanesulfonate phosphate buffer solution and acetonitrile, and the HPLC chart is shown in figure 2 according to the method of example 1.
The following table 2 of peak-out times versus corresponding substances can be obtained according to fig. 2 corresponding to the localization peaks of the respective impurities.
TABLE 2 response time and corresponding substances
As can be seen from fig. 2: the main peak and each impurity peak can be well separated and synchronized, and the peak purity of the main peak meets the regulation.
3. Compound ipratropium bromide solution thermal destruction for inhalation
The inhalation compound ipratropium bromide solution is taken and destroyed for 10 days at 60 ℃, and the HPLC chart is shown in figure 3 according to the method of example 1.
The peak times in fig. 3 are shown in table 3.
TABLE 3 peak time of characteristic peaks of FIG. 3
As can be seen from fig. 3 and the response times of the chromatogram peaks: under the condition of thermal damage, the compound ipratropium bromide solution for inhalation has more degradation products, and the main peak and the adjacent impurity peaks can be well separated and synchronized, and the peak purity of the main peak meets the regulations.
4. Photodisruption of compound ipratropium bromide solution for inhalation
The inhalation compound ipratropium bromide solution is subjected to destruction under 4500Lx light intensity for 10 days, and the HPLC chart is shown in figure 4.
The peak times in fig. 4 are shown in table 4.
Table 4 peak time of characteristic peaks in fig. 4
As can be seen from the response times of the characteristic peaks of fig. 4 and 4: under the condition of light damage, the compound ipratropium bromide solution for inhalation has more degradation products, and the main peak and the adjacent impurity peaks can be well separated and synchronized, and the peak purity of the main peak meets the regulations.
From the above test results, it can be seen that: the detection method can well separate ipratropium bromide, salbutamol sulfate and impurity peaks thereof, and the peak purity of a main peak meets the requirement, so the method can be used for quality control of compound ipratropium bromide solution for inhalation.
EXAMPLE 2 specificity experiments
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is regulated to 2.5 to be a mobile phase A, acetonitrile is used as a mobile phase B, the detection wavelength is 210nm, the column temperature is 35 ℃, and the flow rate is 1.0ml/min.
The gradient elution is set as follows:
1. the compound ipratropium bromide solution blank auxiliary material (from Shandong New era pharmaceutical Co., ltd.) is inhaled, and the HPLC chart is shown in figure 5 according to the method of example 2.
As can be seen from fig. 5: the blank auxiliary material has no interference to the determination of related substances of the sample.
2. Determination of substances related to the compound ipratropium bromide solution for inhalation: a proper amount of compound ipratropium bromide solution for inhalation is taken, and a solution containing 0.2mg/ml of ipratropium bromide is prepared by using a sodium heptanesulfonate phosphate buffer solution-acetonitrile mixed solution, and the HPLC chart is shown in figure 6 according to the method of example 2.
As can be seen from fig. 6: the inspection of relevant substances of the compound ipratropium bromide solution sample for inhalation meets the regulation, and the peak purity of the main peak meets the requirement simultaneously.
Example 3 method durability test
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 215nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate was adjusted to 2.5 to mobile phase A, methanol was used as mobile phase B, and the column temperature was 30℃and the flow rate was 0.9ml/min.
The gradient elution is set as follows:
and preparing a mixed solution of 10 mug/ml of each of ipratropium bromide, salbutamol sulfate, ipratropium bromide impurity C, salbutamol impurity B, salbutamol impurity C, salbutamol impurity D and salbutamol impurity F by using a sodium heptanesulfonate phosphate buffer solution-methanol mixed solution, precisely weighing 25 mug of each mixed solution, and injecting the mixed solution into a liquid chromatograph to record a chromatogram.
Under the condition, the retention time of the main peak of ipratropium bromide is about 33min, the retention time of the main peak of salbutamol sulfate is about 14min, and the separation degree of each impurity is good.
Example 4 method durability test
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is regulated to 2.5 by phosphoric acid to be a mobile phase A, acetonitrile is used as a mobile phase B, the flow rate is 1.1ml/min, the detection wavelength is 210nm, and the column temperature is 40 ℃.
The gradient elution is set as follows:
and preparing a mixed solution of 10 mug/ml of each of ipratropium bromide, salbutamol sulfate, ipratropium bromide impurity C, salbutamol impurity B, salbutamol impurity C, salbutamol impurity D and salbutamol impurity F by using a sodium heptanesulfonate phosphate buffer solution-acetonitrile mixed solution, precisely weighing 25 mug of each mixed solution, and injecting the mixed solution into a liquid chromatograph to record a chromatogram.
Under the condition, the retention time of the main peak of ipratropium bromide is about 31min, the retention time of the main peak of salbutamol sulfate is about 12min, and the separation degree of each impurity is good.
Example 5 method durability test
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is regulated to 2.0 by phosphoric acid to be a mobile phase A, acetonitrile is used as a mobile phase B, the flow rate is 1.0ml/min, the detection wavelength is 210nm, and the column temperature is 35 ℃.
The gradient elution is set as follows:
and preparing a mixed solution of 10 mug/ml of each of ipratropium bromide, salbutamol sulfate, ipratropium bromide impurity C, salbutamol impurity B, salbutamol impurity C, salbutamol impurity D and salbutamol impurity F by using a sodium heptanesulfonate phosphate buffer solution-acetonitrile mixed solution, precisely weighing 25 mug of each mixed solution, and injecting the mixed solution into a liquid chromatograph to record a chromatogram.
Under the condition, the retention time of the main peak of ipratropium bromide is about 32min, the retention time of the main peak of salbutamol sulfate is about 14min, and the separation degree of each impurity is good.
Example 6 method durability test
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is regulated to 3.5 by phosphoric acid to be a mobile phase A, acetonitrile is used as a mobile phase B, the flow rate is 1.1ml/min, the detection wavelength is 205nm, and the column temperature is 35 ℃.
The gradient elution is set as follows:
and preparing a mixed solution of 10 mug/ml of each of ipratropium bromide, salbutamol sulfate, ipratropium bromide impurity C, salbutamol impurity B, salbutamol impurity C, salbutamol impurity D and salbutamol impurity F by using a sodium heptanesulfonate phosphate buffer solution-acetonitrile mixed solution, precisely weighing 25 mug of each mixed solution, and injecting the mixed solution into a liquid chromatograph to record a chromatogram.
Under the condition, the retention time of the main peak of ipratropium bromide is about 32min, the retention time of the main peak of salbutamol sulfate is about 14min, and the separation degree of each impurity is good.
Example 7 impurity correction factor calculation
Instrument and conditions: waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; the pH of the buffer solution containing 0.006mol/L sodium heptanesulfonate is adjusted to 3.5 by phosphoric acid to be a mobile phase A, acetonitrile is used as a mobile phase B, the detection wavelength is 210nm, and the column temperature is 35 ℃.
The gradient elution is set as follows:
mixing ipratropium bromide, salbutamol sulfate, ipratropium bromide impurity C, salbutamol impurity B, salbutamol impurity C, salbutamol impurity D and salbutamol impurity F with sodium heptanesulfonate phosphate buffer solution-acetonitrile to obtain 0.5 μg/ml of each mixed solution, preparing 1 μg/ml, 2 μg/ml, 3 μg/ml, 4 μg/ml, 6 μg/ml, 8 μg/ml and 10 μg/ml of mixed solution by the same method, precisely weighing 25 μl of each mixed solution, injecting into a liquid chromatograph, and recording a chromatogram. The results are shown in Table 3 and Table 4.
TABLE 3 ipratropium bromide and impurities thereof
TABLE 4 salbutamol and its impurities
From the test results in tables 3 to 4, it can be seen that: the detection method can well separate ipratropium bromide and salbutamol sulfate from impurities thereof, and the peak purity of main peaks accords with the regulation.
Comparative example 1:
the liquid chromatography conditions used: LC-10AT liquid chromatograph.
Chromatographic column: phenomenex-C8 (4.6mm.times.250 mm,5 μm) and ultimonate XB-C8 (4.6mm.times.250 mm,5 μm)
Mobile phase: a:0.25% sodium heptanesulfonate solution (29:71) pH was adjusted to 3.2 with phosphoric acid, B: acetonitrile
Sample injection amount: 20 μl of
Detection wavelength: 210nm of
Flow rate: 1.0ml/min
The detection method comprises the following steps:
taking an ipratropium bromide solution for inhalation, and diluting the solution which is prepared by one time by using a mobile phase A solution as a sample solution; precisely measuring 1ml of the sample solution, placing in a 100ml measuring flask, diluting to scale with the mobile phase A solution, and shaking to obtain a control solution.
Taking 20 mu l of control solution, injecting into a liquid chromatograph, adjusting the sensitivity of the instrument to ensure that the main peak height is 20% of the full range, taking 20 mu l of sample solution, injecting into the liquid chromatograph, and recording a chromatogram.
According to the detection result, the condition can detect the ipratropium bromide and related partial impurities thereof, but the information of salbutamol sulfate and the impurities thereof cannot be detected, and the separation degree of the ipratropium bromide detected by the method and the impurities thereof is not good, so the method is not suitable for detecting the related substances of the compound ipratropium bromide for inhalation.
Comparative example 2:
sample preparation methods are consistent as in example 1
Chromatographic conditions:
NertsilODS-3C18 (150 mm. Times.4.6 mm,5 μm) column; mobile phase: 0.01mol.L -1 Potassium dihydrogen phosphate solution (pH is regulated to 2.3 by phosphoric acid) (A) -acetonitrile (B), gradient elution (0-10 min, 5-20% B; 10-15 min,20% B; 15-16 min, 20-5% B; 16-25 min,5% B); flow rate 1.0mL/min -1 The method comprises the steps of carrying out a first treatment on the surface of the Detection wavelength: 220nm (detection of ipratropium bromide) and 276nm (detection of salbutamol sulfate); column temperature: 40 ℃; sample injection amount: 20. Mu.L.
Reference substance solution
Precisely weighing reference substances ipratropium bromide 10mg and salbutamol sulfate 60mg (about equivalent to salbutamol sulfate 50 mg), placing into a 50mL measuring flask, adding water to dissolve and dilute to scale, and shaking to obtain mixed reference substance stock solution; precisely measuring 2.5mL, placing into a 25mL measuring flask, adding water to dilute to a scale, and shaking uniformly to obtain the product.
Test solution
Taking an ipratropium bromide solution for inhalation, precisely measuring 5mL, placing into a 25mL measuring flask, adding the mobile phase A to dilute to a scale, and shaking uniformly to obtain the ipratropium bromide solution.
As a result, only the components of salbutamol sulfate and ipratropium bromide, which are the main components, can be detected from the detection pattern, and the contents of other substances other than the main components cannot be detected, so that the present detection method is not suitable for the detection of substances of interest in ipratropium bromide for inhalation.
Comparative example 3
Waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; and (3) regulating the pH value of the buffer solution containing 0.006mol/L sodium heptanesulfonate to 2.5 to obtain a mobile phase A, and taking acetonitrile as a mobile phase B, wherein the volume ratio of the mobile phase A to the mobile phase B is 90:10, the detection wavelength is 210nm, the column temperature is 35 ℃, the flow rate is 1.0ml/min, the sample injection amount is 25 mu L, and the elution is isocratic.
Both control and test were prepared as in example 1
The method can detect ipratropium bromide and part of impurities thereof, but cannot detect the impurities of salbutamol sulfate.
Comparative example 4
Waters liquid chromatography system, 2998 detector, column: waters Xbridge C18 (150X 4.6mm,3.5 μm); detection wavelength: 210nm; and (3) regulating the pH value of a sodium heptanesulfonate buffer solution to 2.5 by using phosphoric acid with the concentration of 0.006mol/L to obtain a mobile phase A, and using acetonitrile as a mobile phase B, wherein the volume ratio of the mobile phase A to the mobile phase B is 90:10, the detection wavelength is 210nm, the column temperature is 35 ℃, the flow rate is 1.0ml/min, the sample injection amount is 25 μl, and the gradient elution is carried out.
The gradient elution is set as follows:
the preparation of the test and control products was the same as in example 1
By detecting the sample and the control separately by the method of comparative example 4, the detection method can detect a part of the relevant substances and the main component of ipratropium bromide for inhalation, but the separation degree of the relevant substances from the main component is not high, and the method cannot be used for quality control of the relevant substances of ipratropium bromide for inhalation.
Claims (1)
1. A synchronous detection method for five related substances in a compound ipratropium bromide solution for inhalation is characterized in that the related substances are as follows: ipratropium bromide impurity C is 2-phenyl-3-hydroxypropionic acid, salbutamol impurity B is (1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1- (4-hydroxyphenyl) ethanol, salbutamol impurity C is 3-deoxysalbutamol, salbutamol impurity D is 5- [ (1 RS) -2- [ (1, 1-dimethylethyl) amino ] -1-hydroxyethyl ] -2-hydroxyphenyl, salbutamol impurity F is 3,3' - [ oxybis (methylene) ] bis [ alpha- [ [ (1, 1-dimethylethyl) amino ] methyl ] -4-hydroxybenzyl alcohol; the detection method is a high performance liquid chromatography method, and the high performance liquid chromatography method comprises the following chromatographic conditions:
the chromatographic column is Waters Xbridge C18; wherein the filler of the chromatographic column is octadecylsilane chemically bonded silica gel;
the detection wavelength is 210nm;
the flow rate of the mobile phase is 1.0ml/min;
the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is sodium heptanesulfonate phosphate buffer solution, and the mobile phase B is acetonitrile; the pH value of the sodium heptanesulfonate phosphate buffer solution is 2.5, and the concentration of the sodium heptanesulfonate phosphate buffer solution is 0.006mol/L; controlling the column temperature to be 30-40 ℃;
gradient elution is adopted, and the gradient elution is set as follows:
。
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| CN114034782B (en) * | 2021-09-23 | 2024-01-30 | 安徽四环科宝制药有限公司 | Detection method of salbutamol sulfate related substances |
| CN115372502B (en) * | 2022-07-27 | 2023-08-01 | 石家庄四药有限公司 | Method for detecting compound ipratropium bromide medicament for intra-respiratory tract inhalation |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108627597A (en) * | 2018-05-25 | 2018-10-09 | 成都倍特药业有限公司 | A kind of detection method of the salbutamol sulfate in relation to substance |
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| Publication number | Priority date | Publication date | Assignee | Title |
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Non-Patent Citations (1)
| Title |
|---|
| HPLC法测定复方异丙托溴铵吸入溶液两组分的含量;刘斌;《药物分析杂志》;20111231;第31卷(第5期);920-922 * |
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