阅读说明：本技术 一种吸入用复方异丙托溴铵溶液五种有关物质同步检测法 (Synchronous detection method for five related substances of compound ipratropium bromide solution for inhalation ) 是由 张贵民 郭小为 刘阿利 于 2019-03-20 设计创作，主要内容包括：本发明涉及一种吸入用复方异丙托溴铵溶液五种有关物质同步检测法,属于药物质量测定方法技术领域,本发明采用高效液相色谱法进行检测,采用烷基键合硅胶为填料的色谱柱,以含庚烷磺酸钠的磷酸盐缓冲液为流动相A,以有机相为流动相B,检测波长为205-300nm。本发明的检测方法可以将异丙托溴铵和硫酸沙丁胺醇与其它杂质有效分离,具有操作简单,结果全面、准确可靠,专一性强的特点。(The invention relates to a synchronous detection method for five related substances in a compound ipratropium bromide solution for inhalation, which belongs to the technical field of a medicine quality detection method and adopts high performance liquid chromatography for detection, wherein a chromatographic column with alkyl bonded silica gel as a filler is adopted, a phosphate buffer solution containing sodium heptanesulfonate is taken as a mobile phase A, an organic phase is taken as a mobile phase B, and the detection wavelength is 205-300 nm. The detection method can effectively separate ipratropium bromide, salbutamol sulfate and other impurities, and has the characteristics of simple operation, comprehensive, accurate and reliable result and strong specificity.)

1. A synchronous detection method for five related substances of 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 (1RS) -2- [ (1, 1-dimethylethyl) amino ] -1- (4-hydroxyphenyl) ethanol, salbutamol impurity C is 3-deoxysalbutamol, salbutamol impurity D is 5- [ (1RS) -2- [ (1, 1-dimethylethyl) amino ] -1-hydroxyethyl ] -2-hydroxyphenyl, and salbutamol impurity F is 3,3' - [ oxybis (methylene) ] bis [ alpha- [ [ (1, 1-dimethylethyl) amino ] methyl ] -4-hydroxybenzyl alcohol; the detection method is high performance liquid chromatography and adopts gradient elution.

2. The method according to claim 1, characterized in that the high performance liquid chromatography comprises the following chromatographic conditions:

the filler of the chromatographic column is alkyl bonded silica gel;

the detection wavelength is 205nm-300 nm;

the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is a sodium heptanesulfonate phosphoric acid 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 ℃.

3. The method of claim 1, wherein the gradient elution is configured to:

4. the method of claim 1, wherein the gradient elution is configured to:

5. the method of claim 2, wherein the flow rate of the mobile phase is 1.0 ml/min.

6. The method of claim 2, wherein the packing material of the chromatography column is octadecylsilane bonded silica or octylsilane bonded silica.

7. The process according to claim 2, characterized in that the organic phase in the mobile phase is methanol or acetonitrile, preferably acetonitrile.

8. The method of claim 2, wherein the detection wavelength is 210 nm.

9. The method according to claim 2, wherein the pH value of the sodium heptanesulfonate phosphate buffer is 2.0-3.5, preferably 2.5.

10. The method according to claim 2, wherein the concentration of the sodium heptanesulfonate phosphate buffer is 0.006 mol/L.

Technical Field

The invention relates to the technical field of medicine quality determination 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 the main components are ipratropium bromide and salbutamol sulfate. The product is suitable for patients in need of combination of multiple bronchodilators, and can be used for treating reversible bronchospasm related to airway obstructive diseases. Wherein, the ipratropium bromide and the salbutamol are superposed to act on the muscarinic and beta 2 adrenergic receptors of the lung to generate the effect of bronchiectasis, and the curative effect is better than that of single administration.

Determination of related substances [ J ] in ipratropium bromide solution for inhalation by reversed phase high performance liquid chromatography, Chongqing medicine, 2007(11):1067-1068 ″, which discloses that chromatographic columns are phenomenex-C8(4.6 mm. times.250 mm,5 μm) and ultime XB-C8(4.6 mm. times.250 mm,5 μm), mobile phase A is acetonitrile-0.25% sodium heptanesulfonate solution (29:71) is adjusted to pH 3.2 by phosphoric acid, and mobile phase B is acetonitrile; the detection wavelength is 210 nm; the flow rate is 1.0 ml/min; the detection condition of the sample volume of 20 mul is used for detecting related substances in the ipratropium bromide solution, but salbutamol sulfate and related impurities can not be detected.

Literature' HPLC method for determining contents of two components of compound ipratropium bromide inhalation solution [ J]1067-1068' the method establishes a high performance liquid chromatography to simultaneously determine the contents of two components of ipratropium bromide and salbutamol sulfate in a compound ipratropium bromide inhalation solution, adopts an Inertsil ODS-3C18(150mm × 4.6.6 mm,5 μm) chromatographic column, and the mobile phase is 0.01mol.L^-1Potassium dihydrogen phosphate solution (pH 2.3 adjusted with phosphoric acid) (A) -acetonitrile (B), gradient elution, flow rate 1.0mL.min^-1The detection wavelengths are 220nm (for detecting ipratropium bromide) and 276nm (for detecting salbutamol), and the column temperature is 40 ℃. As a result: the sample injection amounts of ipratropium bromide and salbutamol are respectively 0.01-0.2 mg^-1And 0.05-1.0 mg.mL^-1The linear relationship was good within the range (r was 0.9999 and n was 5), and the recovery rate (n was 9) was 100.2% and 100.3%, respectively. Although the method detects the contents of the two components of the compound salbutamol, the method does not further detect the contents of related substances and has complex components between the related substances, and the method cannot obviously separate the related substances from the main components and cannot ensure the measurement sensitivity and specificity.

Because the related substance determination standard 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 various related substances of the compound ipratropium bromide solution for inhalation at one time is very important.

Disclosure of Invention

In view of the defects in the prior art, the invention aims to provide a synchronous detection method for five related substances in a compound ipratropium bromide solution for inhalation by improving and optimizing the aspects of gradient, mobile phase, column temperature, flow rate and the like of gradient elution, and the method can be used for quality control in the production of the compound ipratropium bromide solution for inhalation. The preparation operation of the mobile phase in the method is simple, the analysis method is stable, the sensitivity is high, the peak pattern is good, and various related substances can be detected at one time.

In order to achieve the purpose of the invention, the inventor finally and preferably selects the following technical scheme through a large number of experiments and creative efforts:

a synchronous detection method for five related substances of a compound ipratropium bromide solution for inhalation is disclosed, wherein the five related substances of the compound ipratropium bromide solution for inhalation are as follows: ipratropium bromide impurity C (2-phenyl-3-hydroxypropionic acid), salbutamol impurity B ((1RS) -2- [ (1, 1-dimethylethyl) amino ] -1- (4-hydroxyphenyl) ethanol), salbutamol impurity C (3-deoxysalbutamol), salbutamol impurity D (5- [ (1RS) -2- [ (1, 1-dimethylethyl) amino ] -1-hydroxyethyl ] -2-hydroxybenzene), salbutamol impurity F (3,3' - [ oxybis (methylene) ] bis [ α - [ [ (1, 1-dimethylethyl) amino ] methyl ] -4-hydroxybenzyl alcohol), said assay being high performance liquid chromatography, employing gradient elution.

Preferred chromatographic conditions for high performance liquid chromatography are:

the filler of the chromatographic column is alkyl bonded silica gel;

the detection wavelength is 205-300 nm;

the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A is a sodium heptanesulfonate phosphoric acid buffer solution, and the mobile phase B is an organic phase;

setting the flow rate of a 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 can be set as follows:

more preferred gradient elution may be set as:

the preferred alkyl-bonded silica gel for the filler is octadecylsilane bonded silica gel or octylsilane 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, and more preferably, the sodium heptanesulfonate phosphate buffer is 0.006 mol/L.

The pH value of the sodium heptanesulfonate phosphate buffer solution is 2.0-3.5, and more preferably, the pH value of the sodium heptanesulfonate phosphate buffer solution is 2.5.

Preferably, the flow rate of the mobile phase in the process of the invention is set at 1.0 ml/min.

It should be noted that the method of the present invention is described in detail in the specific examples, and the method of the present invention is verified, and the results show that the detection method of the present invention has the following unexpected effects compared with the prior art:

(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 by 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 condition replacement in the detection process, greatly improves the working efficiency and reduces the cost.

(3) The method has the advantages of high separation degree, good repeatability and durability, short analysis time, simple operation and stable and reliable result.

Therefore, the invention can be used for controlling the quality of the compound ipratropium bromide solution for inhalation and provides effective guarantee for the quality of the final finished product.

Drawings

FIG. 1 HPLC chromatogram of compound ipratropium bromide solution mixed impurity reference substance for inhalation

FIG. 2 HPLC chromatogram of related substances of compound ipratropium bromide solution for inhalation

FIG. 3 thermal destruction HPLC chromatogram of compound ipratropium bromide solution for inhalation

FIG. 4 photo-destructive HPLC profile of a solution of ipratropium bromide for inhalation

FIG. 5 HPLC chromatogram of blank adjuvant of compound ipratropium bromide solution for inhalation

FIG. 6 HPLC chromatogram of related substances of compound ipratropium bromide solution for inhalation

Detailed Description

The following are specific examples of the present invention and further describe the technical solutions of the present invention, but the scope of the present invention is not limited to these examples. All changes, modifications and equivalents that do not depart from the spirit of the invention are intended to be included within the scope thereof.