CN110579542B - Method for measuring ranitidine hydrochloride related substances by high performance liquid chromatography - Google Patents
Method for measuring ranitidine hydrochloride related substances by high performance liquid chromatography Download PDFInfo
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Abstract
The invention relates to the field of drug detection, in particular to a method for determining related substances of ranitidine hydrochloride by high performance liquid chromatography. The method adopts octadecylsilane chemically bonded silica as a filler; the mobile phase comprises a mobile phase A and a mobile phase B, wherein the mobile phase A and the mobile phase B are mixed solutions of modified phosphate buffer solution and acetonitrile, and the pH value of the modified phosphate buffer solution is 6.70 +/-0.05; the flow rate of the mobile phase is 1.1-1.3 ml/min; gradient elution was used. The method can separate related substances of ranitidine hydrochloride in a high performance liquid chromatogram; and the sensitivity and accuracy of detection of each component are further improved by optimizing the conditions. The method can better control the quality of the ranitidine hydrochloride, has high analysis speed, good specificity and high repeatability, is convenient for detecting and monitoring the quality of the ranitidine hydrochloride, and is beneficial to the safe popularization and application of the ranitidine hydrochloride.
Description
Technical Field
The invention relates to the field of drug detection, in particular to a method for determining related substances of ranitidine hydrochloride by high performance liquid chromatography.
Background
Ranitidine hydrochloride is histamine H2 receptor blocking agent, and can inhibit basic gastric acid secretion, stimulated gastric acid secretion and pepsin secretion. The acid inhibition strength of the compound is 5-8 times stronger than that of cimetidine. In 1977, ranitidine (AH19065) was first developed by ale n & Hanburys, inc, of john brad shoji, a subsidiary of gratin smith, and was first marketed in the uk at 10 months in 1981.
However, the ranitidine hydrochloride is easy to generate impurities when used as a medicine, 12 process impurities and degradation impurities are known, and the harm of the ranitidine hydrochloride to a human body cannot be evaluated, so that the impurities and the ranitidine are effectively separated, the amount of the impurities is controlled, and the risk is reduced. At present, the prior art does not disclose the determination method of 12 known impurities (namely, the impurity A-the impurity L described below) of ranitidine hydrochloride, so that the determination method of various processes and degradation impurities of ranitidine hydrochloride needs to be provided.
In view of the above, the present invention is particularly proposed.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides a method for determining related substances of ranitidine hydrochloride by using a high performance liquid chromatography.
Specifically, the method adopts octadecylsilane chemically bonded silica as a filler; the mobile phase consists of a mobile phase A and a mobile phase B, wherein the mobile phase A is prepared from a modified phosphate buffer solution and acetonitrile according to a volume ratio of 97-99: 1-3, wherein the mobile phase B is prepared from a modified phosphate buffer solution and acetonitrile according to a volume ratio of 77-79: 21-23, wherein the pH value of the modified phosphate buffer solution is 6.70 +/-0.05; adopting gradient elution, wherein the total volume of the mobile phase is 100 percent,
in 0-15 min, the volume of the mobile phase A is decreased from 100% to 0%, and the volume of the mobile phase B is increased from 0% to 100%;
in 15-23 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 23-24 min, the volume of the mobile phase A is increased from 0% to 100%, and the volume of the mobile phase B is decreased from 100% to 0%;
and at 24-30 min, the volume of the mobile phase A is 100%, and the volume of the mobile phase B is 0%.
The invention discovers that when the high performance liquid chromatography is adopted to detect the impurities in the ranitidine hydrochloride, the selection of the mobile phase has great influence on the separation of the impurities, and when the mobile phase A and the mobile phase B with fixed proportion are adopted to detect, the impurities are difficult to be well separated; it has been found through a number of attempts that ranitidine hydrochloride can be effectively separated from various impurities while shortening the running time when the elution method is adopted.
Furthermore, the invention discovers that when the pH value of the modified phosphate buffer solution is 6.70 +/-0.05, the separation effect is better; not only can separate ranitidine hydrochloride and 12 impurities, but also can shorten the separation time.
The related substances of the ranitidine hydrochloride comprise ranitidine hydrochloride and impurities, wherein the impurities are selected from one or more of ranitidine hydrochloride impurity A, impurity B, impurity C, impurity D, impurity E, impurity F, impurity G, impurity H, impurity I, impurity J, impurity K and impurity L.
The method of the invention can separate the 12 impurities simultaneously and realize ideal separation degree.
In order to further improve the accuracy and sensitivity of detection, the invention optimizes other conditions of the high performance liquid chromatography, and obtains the following preferred schemes (combining the preferred schemes can obtain the preferred embodiment of the invention):
preferably, the flow rate of the mobile phase is 1.1 to 1.3 ml/min.
Preferably, triethylamine is added into the phosphate buffer solution to prepare the modified phosphate buffer solution.
Preferably, the triethylamine accounts for 0.04-0.06% of the modified phosphate buffer solution.
Preferably, the preparation method of the modified phosphate buffer solution is as follows: dissolving phosphoric acid in water, adding triethylamine and 50% sodium hydroxide solution, and adding water to a constant volume; the pH is adjusted to 6.70. + -. 0.05 with phosphoric acid or 50% sodium hydroxide solution.
Preferably, the specification of the chromatographic column is 4.6mm × 250mm, 5 μm; preferably Welch-Xtimate-C18, 4.6 mm. times.250 mm, 5 μm.
In the present invention, preferably, a Yueuxu peak suppressor is added before the column.
As a preferred embodiment of the present invention, the method of the present invention employs the following conditions for measurement:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler; the specification is Welch-Xtimate-C18, 4.6mm multiplied by 250mm, 5 μm;
mobile phase A: the volume ratio is 97-99: 1-3 of a mixed solution of a modified phosphate buffer solution and acetonitrile;
mobile phase B: the volume ratio is 77-79: 21-23 of a mixed solution of a modified phosphate buffer solution and acetonitrile;
flow rate: 1.1-1.3 ml/min;
column temperature: 23-27 ℃;
detection wavelength: 228 nm-232 nm;
adopting gradient elution, wherein the total volume of the mobile phase is 100 percent,
in 0-15 min, the volume of the mobile phase A is decreased from 100% to 0%, and the volume of the mobile phase B is increased from 0% to 100%;
in 15-23 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 23-24 min, the volume of the mobile phase A is increased from 0% to 100%, and the volume of the mobile phase B is decreased from 100% to 0%;
and at 24-30 min, the volume of the mobile phase A is 100%, and the volume of the mobile phase B is 0%.
Preferably, the ranitidine hydrochloride is diluted to 0.8-1.2 mg/mL by using a solvent to be used as a test solution; preferably 1 mg/ml.
Preferably, the sample solution is diluted to 0.8-1.2 mu g/ml by a solvent to be used as a control solution; preferably 1. mu.g/ml.
Preferably, the ranitidine hydrochloride reference substance and each ranitidine hydrochloride impurity reference substance are diluted by a solvent to be used as a system applicability solution;
in the system applicability solution, the concentration of a ranitidine hydrochloride reference substance is 0.8-1.2 mg/ml, and the concentration of each ranitidine hydrochloride impurity reference substance is 0.8-1.2 mu g/ml; preferably, the concentration of the ranitidine hydrochloride reference substance is 1mg/ml, and the concentration of each ranitidine hydrochloride impurity reference substance is 1 mu g/ml.
Preferably, the solvent is water.
The invention has the beneficial effects that:
the method provided by the invention can separate related substances of ranitidine hydrochloride (comprising ranitidine hydrochloride and 12 impurities) in a high performance liquid chromatogram; and the sensitivity and accuracy of detection of each component are further improved by optimizing the conditions. The method can better control the quality of the ranitidine hydrochloride, has high analysis speed, good specificity and high repeatability, is convenient for detecting and monitoring the quality of the ranitidine hydrochloride, and is beneficial to the safe popularization and application of the ranitidine hydrochloride.
Drawings
FIG. 1 is a chromatogram of a system suitability solution in Experimental example 1.
FIG. 2 is a chromatogram of a system suitability solution of comparative example 1.
Detailed Description
The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Example 1
The embodiment provides a method for measuring related substances of ranitidine hydrochloride by high performance liquid chromatography, which adopts the following conditions for measurement:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler; the specification of the chromatographic column is Welch-Xtimate-C18, 4.6mm × 250mm, 5 μm; adding a Yueuxu peak suppressor in front of the column;
mobile phase A: the volume ratio is 98:2 (6.8 ml of phosphoric acid is dissolved in 1900ml of water, 1ml of triethylamine and 8.6ml of 50% sodium hydroxide solution are added, the water is added to reach 2000ml, and the pH value is adjusted to 6.70 +/-0.05 by using phosphoric acid or 50% sodium hydroxide solution) and acetonitrile;
mobile phase B: the volume ratio is 78: 22, a mixed solution of a modified phosphate buffer solution and acetonitrile;
flow rate: 1.2 ml/min;
column temperature: 25 ℃;
detection wavelength: 230 nm;
adopting gradient elution, wherein the total volume of the mobile phase is 100 percent,
in 0-15 min, the volume of the mobile phase A is decreased from 100% to 0%, and the volume of the mobile phase B is increased from 0% to 100%;
in 15-23 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 23-24 min, the volume of the mobile phase A is increased from 0% to 100%, and the volume of the mobile phase B is decreased from 100% to 0%;
and at 24-30 min, the volume of the mobile phase A is 100%, and the volume of the mobile phase B is 0%.
Ranitidine hydrochloride was diluted to 1mg/mL with a solvent (water) as a test solution.
The test solution was diluted to 1. mu.g/ml with a solvent (water) as a control solution.
Diluting a ranitidine hydrochloride reference substance, a ranitidine hydrochloride impurity A reference substance, an impurity B reference substance, an impurity C reference substance, an impurity D reference substance, an impurity E reference substance, an impurity F reference substance, an impurity G reference substance, an impurity H reference substance, an impurity I reference substance, an impurity J reference substance, an impurity K reference substance and an impurity L reference substance by using a solvent (water) to serve as a system applicability solution;
in the system applicability solution, the concentration of a ranitidine hydrochloride reference substance is 1mg/ml, and the concentration of each ranitidine hydrochloride impurity reference substance is 1 mu g/m.
The specific preparation process of each solution and the test verification process and the result of the method are shown in experimental examples 1-3.
Experimental example 1 System suitability test
Preparation of a test solution: taking about 100mg of ranitidine hydrochloride, precisely weighing, placing in a 100ml measuring flask, adding a proper amount of water to dissolve, diluting with water to a scale, and shaking up to be used as a test solution.
Preparation of control solution: precisely measuring a proper amount of a test solution, and quantitatively diluting the test solution with a solvent to prepare a solution containing 1 mu g of ranitidine hydrochloride in 1ml as a control solution.
Preparation of system suitability solution: the method comprises the following steps of precisely weighing appropriate amounts of a ranitidine hydrochloride impurity A reference substance, an impurity B reference substance, an impurity C reference substance, an impurity D reference substance, an impurity E reference substance, an impurity F reference substance, an impurity G reference substance, an impurity H reference substance, an impurity I reference substance, an impurity J reference substance, an impurity K reference substance, an impurity L reference substance and a ranitidine hydrochloride reference substance, adding a solvent to dissolve and dilute into a mixed solution containing 1 mu G of each impurity reference substance and 1mg of the ranitidine hydrochloride reference substance per 1ml, and taking the mixed solution as a system applicability solution.
Preparing an impurity positioning solution: the method comprises the following steps of precisely weighing a proper amount of each of a ranitidine hydrochloride impurity A reference substance, an impurity B reference substance, an impurity C reference substance, an impurity D reference substance, an impurity E reference substance, an impurity F reference substance, an impurity G reference substance, an impurity H reference substance, an impurity I reference substance, an impurity J reference substance, an impurity K reference substance, an impurity L reference substance and a ranitidine hydrochloride reference substance, respectively adding a solvent (water) to dissolve and dilute the materials to obtain 10 mu G/mL solution serving as each impurity positioning solution;
and (3) determination: octadecylsilane chemically bonded silica is used as a filler for a chromatographic column, a mobile phase A is modified phosphate buffer solution (6.8 ml of phosphoric acid is dissolved in 1900ml of water, 1ml of triethylamine and 8.6ml of 50% sodium hydroxide solution are added, water is added to the mixture to reach 2000ml, the pH value is adjusted to 6.70 +/-0.05) -acetonitrile (98:2) by using phosphoric acid or 50% sodium hydroxide solution, a mobile phase B is modified phosphate buffer solution-acetonitrile (78: 22), the column temperature is 25 ℃, the flow rate is 1.2ml per minute, the detection wavelength is 230nm, and gradient elution is adopted.
And precisely measuring 10 mu l of each of the positioning solution and the system applicability solution, injecting into a high performance liquid chromatograph, and recording a chromatogram. The results are shown in Table 1, and the chromatographic chart of the applicability of the ranitidine hydrochloride system is shown in FIG. 1.
TABLE 1 specificity-localization test results
And (4) conclusion: the solvent does not interfere the determination of related substances of known impurities in the test solution, the impurities are well separated from the main peak, and the tailing factors and the theoretical pedal number meet the determination requirements of the related substances.
Experimental example 2 Linear and Range testing
Solvent: water (W)
Linear stock solution: taking about 10mg of each of a ranitidine hydrochloride impurity A reference substance, an impurity B reference substance, an impurity C reference substance, an impurity D reference substance, an impurity E reference substance, an impurity F reference substance, an impurity G reference substance, an impurity H reference substance, an impurity I reference substance, an impurity J reference substance, an impurity K reference substance, an impurity L reference substance and a ranitidine hydrochloride reference substance, respectively placing the ranitidine hydrochloride impurity A reference substance, the impurity B reference substance, the impurity C reference substance, the impurity D reference substance, the impurity E reference substance, the impurity F reference substance, the impurity G reference substance, the impurity H reference substance, the impurity I reference substance, the impurity J reference substance, the impurity K reference substance, the impurity L reference substance and the ranitidine hydrochloride reference substance into 100ml measuring bottles, adding a proper amount of solvent for ultrasonic dissolution and dilution to scale, shaking up uniformly, taking the obtained solution as each storage solution, precisely taking a proper amount, and diluting to prepare a series of linear solutions.
Precisely measuring 10 μ l of each solution, injecting into a liquid chromatograph, and recording chromatogram; the results are shown in Table 2.
TABLE 2 Linear and Range test results
And (4) conclusion:
(1) the impurity A is in the range of 0.050 mu g/ml-9.072 mu g/ml (equivalent to 0.005% -0.907% of the concentration of the sample), the linear regression equation is that y is 20284.8518x +379.9061r is 0.9999, and the linear regression is remarkable.
(2) The impurity B is in the range of 0.099 μ g/ml to 2.966 μ g/ml (corresponding to 0.010% to 0.297% of the sample concentration), and the linear regression equation is that y is 12912.0761x +328.7296r is 0.9995, and the linear regression is significant.
(3) The impurity C is in the range of 0.050 μ g/ml to 3.000 μ g/ml (equivalent to 0.005% to 0.300% of the concentration of the test article), the linear regression equation is that y is 24143.1315x +4.6672r is 0.9996, and the linear regression is remarkable.
(4) The impurity D is in the range of 0.047 mug/ml-2.830 mug/ml (equivalent to 0.005% -0.283% of the concentration of the sample), the linear regression equation is that y is 22414.7354x +207.1704r is 0.9998, and the linear regression is remarkable.
(5) The impurity E is in the range of 0.049 μ g/ml to 2.912 μ g/ml (corresponding to 0.005% to 0.291% of the concentration of the test sample), the linear regression equation is that y is 27979.1928x +104.8356r is 0.9997, and the linear regression is significant.
(6) The impurity F is in the range of 0.020 mu g/ml-2.758 mu g/ml (equivalent to 0.002% -0.276% of the concentration of the sample), the linear regression equation is that y is 61023.8172 x-1631.8976 r is 0.9996, and the linear regression is remarkable.
(7) The impurity G is in the range of 0.050 μ G/ml to 3.000 μ G/ml (corresponding to 0.005% to 0.300% of the sample concentration), the linear regression equation is y 21414.0623 x-278.4059 r 0.9995, and the linear regression is remarkable.
(8) The impurity H is in the range of 0.049 μ g/ml to 2.966 μ g/ml (corresponding to 0.005% to 0.297% of the sample concentration), and the linear regression equation is that y is 10730.8585x +249.6983r is 0.9998, and the linear regression is significant.
(9) The impurity I is in the range of 0.037 μ g/ml to 2.801 μ g/ml (equivalent to 0.004% to 0.280% of the concentration of the test article), the linear regression equation is that y is 26377.0006x +12.8052r is 1.0000, and the linear regression is remarkable.
(10) The impurity J is in the range of 0.074 μ g/ml to 4.459 μ g/ml (equivalent to 0.007% to 0.446% of the concentration of the test article), the linear regression equation is that y is 10846.1499x +98.3079r is 0.9997, and the linear regression is remarkable.
(11) The impurity K is in the range of 0.103-3.078 mug/ml (equivalent to 0.010-0.308% of the concentration of the sample), the linear regression equation is that y is 9170.2334x +30.4182r is 0.9997, and the linear regression is remarkable.
(12) The impurity L is in the range of 0.038-2.866 mug/ml (equivalent to 0.004-0.287% of the concentration of the sample), the linear regression equation is that y is 19689.1161x +85.7104r is 0.9998, and the linear regression is remarkable.
(13) The linear regression equation of ranitidine hydrochloride is that y is 22430.5661 x-402.7950 r is 0.9999 in the range of 0.055 mu g/ml-3.233 mu g/ml (equivalent to 0.006% -0.323% of the concentration of the test sample), and the linear regression is obvious.
Experimental example 3 recovery test
Solvent: water;
impurity reference substance: precisely weighing about 10mg each of ranitidine hydrochloride impurity A, impurity B, impurity C, impurity D, impurity E, impurity F, impurity G, impurity H, impurity I, impurity J, impurity K and impurity L, respectively placing the ranitidine hydrochloride impurity A, the impurity B, the impurity C, the impurity D, the impurity E, the impurity F, the impurity G, the impurity H, the impurity I, the impurity J, the impurity K and the impurity L into 100ml measuring bottles, dissolving the ranitidine hydrochloride impurity A, the impurity B, the impurity C, the impurity D, the impurity E, the impurity F, the impurity G, the impurity H, the impurity I, the impurity J, the impurity K and the impurity L by using a diluent, fixing the volume and shaking up the ranitidine hydrochloride impurity A, the impurity B, the impurity C, the impurity D, the impurity E, the impurity F, the impurity G, the impurity H, the impurity I, the impurity J, the impurity K and the impurity L to obtain reference substance stock solutions of each impurity. Precisely measuring 3ml of impurity A reference substance stock solution, 1.5ml of impurity J reference substance stock solution and 1ml of each of other impurity stock solutions, placing the impurity A reference substance stock solutions, the impurity J reference substance stock solutions and the other impurity stock solutions in the same 100ml measuring flask, diluting the impurity A reference substance stock solutions to scales by using a diluent, and shaking up the impurity A reference substance stock solutions and the impurity J reference substance stock solutions to obtain the impurity A reference substance stock solutions; 2 parts are prepared in parallel.
Preparation of an accuracy solution:
50% accuracy solution: taking a proper amount of ranitidine hydrochloride (about equivalent to 200mg of ranitidine), precisely weighing, placing in a 200ml measuring flask, adding a proper amount of water to dissolve, precisely adding 3ml of impurity A reference substance stock solution, 1.5ml of impurity J reference substance stock solution and 1ml of other impurity reference substance stock solutions, adding a diluent to dilute to a scale, and shaking uniformly to obtain the ranitidine hydrochloride; 3 parts are prepared in parallel.
100% accuracy solution: taking a proper amount of ranitidine hydrochloride (about equivalent to 100mg of ranitidine), precisely weighing, placing into a 100ml measuring flask, adding a proper amount of water to dissolve, precisely adding 3ml of impurity A reference substance stock solution, 1.5ml of impurity J reference substance stock solution and 1ml of other impurity reference substance stock solutions, adding a diluent to dilute to a scale, and shaking uniformly to obtain the ranitidine hydrochloride; 3 parts are prepared in parallel.
200% accuracy solution: taking a proper amount of ranitidine hydrochloride (about equivalent to 50mg of ranitidine), precisely weighing, placing in a 50ml measuring flask, adding a proper amount of water to dissolve, precisely adding 3ml of impurity A reference substance stock solution, 1.5ml of impurity J reference substance stock solution and 1ml of other impurity reference substance stock solutions, adding a diluent to dilute to a scale, and shaking uniformly to obtain the ranitidine hydrochloride; 3 parts are prepared in parallel.
Preparing a background solution:
taking a proper amount of ranitidine hydrochloride (about equivalent to 100mg of ranitidine), placing the ranitidine hydrochloride into a 100ml measuring flask, adding water to dissolve and dilute the ranitidine hydrochloride to a scale, shaking up, filtering, and taking a subsequent filtrate as a test solution (newly prepared for clinical use);
the solutions were measured precisely at 10. mu.l each and injected into a liquid chromatograph, and the results are shown in tables 3 to 15.
Table 3 verification of related materials method-accuracy background solution results
| Name (R) | Peak area (A) | Content (%) |
| Impurity A | 10688 | 0.06 |
| Impurity B | Not detected out | / |
| Impurity C | Not detected out | / |
| Impurity D | Not detected out | / |
| Impurity E | Not detected out | / |
| Impurity F | Not detected out | / |
| Impurity G | Not detected out | / |
| Impurity H | Not detected out | / |
| Impurity I | Not detected out | / |
| Impurity J | Not detected out | / |
| Impurity K | Not detected out | / |
| Impurity L | Not detected out | / |
Impurity A is detected in the background solution, and the background amount is required to be deducted when the analytical method of the product is used for accurately calculating.
TABLE 4 verification of the substance method-results on recovery of impurity A
Table 5 verification of related materials methods-results on recovery of impurity B
TABLE 6 verification of related substances method-results on recovery of impurity C
TABLE 7 verification of related substances method-results on recovery of impurity D
TABLE 8 verification of related substances methods-results on recovery of impurity E
TABLE 9 verification of related substances method-results on recovery of impurity F
TABLE 10 verification of related materials methods-impurity G recovery results
TABLE 11 verification of related materials methods-results on recovery of impurity H
TABLE 12 verification of related substances methods-results on recovery of impurity I
TABLE 13 verification of related substances methods-results on recovery of impurity J
TABLE 14 verification of related materials methods-results on recovery of impurity K
TABLE 15 verification of related materials methods-results on recovery of impurity L
And (4) conclusion: the impurity recovery test results show that the recovery rates of 9 accurate samples of the impurity A, the impurity B, the impurity C, the impurity D, the impurity E, the impurity F, the impurity G, the impurity H, the impurity I, the impurity J, the impurity K and the impurity L are all between 90% and 108%, the average recovery rates are respectively 102.5%, 99.2%, 101.6%, 100.1%, 101.0%, 98.7%, 103.4%, 99.0%, 101.1%, 102.9%, 103.0% and 100.0, and tables 4 to 15 show that the recovery rates among impurity groups meet the determination requirements of the product on various known impurities, and the method has good accuracy.
Comparative example 1
The comparative example provides a method for determining related substances of ranitidine hydrochloride by high performance liquid chromatography, and the method is only different from the method in example 1 in that:
a chromatographic column: octadecylsilane bonded silica gel as a packing (Kromasil C184.6 x 150mm, 5 μm or equivalent performance column);
mobile phase A: the volume ratio is 98:2 (a mixture of 6.8ml of phosphoric acid dissolved in 1900ml of water, 8.6ml of 50% sodium hydroxide solution, 2000ml of water, pH adjusted to 7.10. + -. 0.05 with phosphoric acid or 50% sodium hydroxide solution) and acetonitrile;
mobile phase B: the volume ratio is 78: 22 of phosphate buffer solution and acetonitrile;
flow rate: 1.5 ml/min;
column temperature: 35 ℃ is carried out.
The applicability chromatogram of the ranitidine hydrochloride system is shown in figure 2; as can be seen from fig. 2, comparative example 1 failed to effectively separate impurities.
Experiments prove that the impurity tailing can be effectively improved by adding triethylamine; after the pH value is adjusted, the separation degree between the main peak and the degraded impurities is improved; the column temperature is increased, and the separation degree among impurities is improved; the chromatographic condition of the invention can effectively separate impurities, impurities and main peaks.
Although the invention has been described in detail hereinabove by way of general description, specific embodiments and experiments, it will be apparent to those skilled in the art that many modifications and improvements can be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. A method for measuring related substances of ranitidine hydrochloride by high performance liquid chromatography is characterized in that octadecylsilane chemically bonded silica is used as a filling agent; the mobile phase consists of a mobile phase A and a mobile phase B, wherein the mobile phase A is prepared from a modified phosphate buffer solution and acetonitrile according to a volume ratio of 97-99: 1-3, wherein the mobile phase B is prepared from a modified phosphate buffer solution and acetonitrile according to a volume ratio of 77-79: 21-23, wherein the pH value of the modified phosphate buffer solution is 6.70 +/-0.05; adopting gradient elution, wherein the total volume of the mobile phase is 100 percent,
in 0-15 min, the volume of the mobile phase A is decreased from 100% to 0%, and the volume of the mobile phase B is increased from 0% to 100%;
in 15-23 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 23-24 min, the volume of the mobile phase A is increased from 0% to 100%, and the volume of the mobile phase B is decreased from 100% to 0%;
in 24-30 min, the volume of the mobile phase A is 100%, and the volume of the mobile phase B is 0%;
the method is used for detecting ranitidine hydrochloride and 12 impurities thereof, wherein the impurities are impurity A, impurity B, impurity C, impurity D, impurity E, impurity F, impurity G, impurity H, impurity I, impurity J, impurity K and impurity L, and the structural formulas of the impurities are as follows:
adding triethylamine into a phosphate buffer solution to prepare the modified phosphate buffer solution, wherein the triethylamine accounts for 0.04-0.06% of the modified phosphate buffer solution; the column was Welch-Xtimate-C18, 4.6mm X250 mm, 5 μm.
2. The method according to claim 1, wherein the flow rate of the mobile phase is 1.1 to 1.3 ml/min.
3. The method according to claim 1, characterized in that the following conditions are used for the determination:
a chromatographic column: octadecylsilane chemically bonded silica is used as a filler; the specification is Welch-Xtimate-C18, 4.6mm multiplied by 250mm, 5 μm;
mobile phase A: the volume ratio is 97-99: 1-3 of a mixed solution of a modified phosphate buffer solution and acetonitrile;
mobile phase B: the volume ratio is 77-79: 21-23 of a mixed solution of a modified phosphate buffer solution and acetonitrile;
flow rate: 1.1-1.3 ml/min;
column temperature: 23-27 ℃;
detection wavelength: 228 nm-232 nm;
adopting gradient elution, wherein the total volume of the mobile phase is 100 percent,
in 0-15 min, the volume of the mobile phase A is decreased from 100% to 0%, and the volume of the mobile phase B is increased from 0% to 100%;
in 15-23 min, the volume of the mobile phase A is 0%, and the volume of the mobile phase B is 100%;
at 23-24 min, the volume of the mobile phase A is increased from 0% to 100%, and the volume of the mobile phase B is decreased from 100% to 0%;
and at 24-30 min, the volume of the mobile phase A is 100%, and the volume of the mobile phase B is 0%.
4. The method according to any one of claims 1 to 3, wherein the sample solution is prepared by diluting ranitidine hydrochloride with a solvent to 0.8 to 1.2 mg/mL.
5. The method of claim 4, wherein the sample solution is diluted with a solvent to 0.8-1.2 μ g/ml as a control solution.
6. The method according to any one of claims 1 to 3 or 5, wherein a ranitidine hydrochloride reference substance and each ranitidine hydrochloride impurity reference substance are diluted with a solvent as a system suitability solution;
in the system applicability solution, the concentration of a ranitidine hydrochloride reference substance is 0.8-1.2 mg/ml, and the concentration of each ranitidine hydrochloride impurity reference substance is 0.8-1.2 mu g/ml.
7. The method of claim 4, wherein the ranitidine hydrochloride control and each of the ranitidine hydrochloride contaminant controls are diluted with a solvent as a system compatible solution;
in the system applicability solution, the concentration of a ranitidine hydrochloride reference substance is 0.8-1.2 mg/ml, and the concentration of each ranitidine hydrochloride impurity reference substance is 0.8-1.2 mu g/ml.
8. The method of claim 4, wherein the solvent is water.
9. The method of claim 6, wherein the solvent is water.
10. The method according to claim 5 or 7, wherein the solvent is water.
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| CN111272893A (en) * | 2020-02-28 | 2020-06-12 | 佛山手心制药有限公司 | Detection method and application of ranitidine hydrochloride |
| CN112526005A (en) * | 2020-10-14 | 2021-03-19 | 江苏汉斯通药业有限公司 | High performance liquid chromatography analysis method for nitrosamines in ranitidine drugs |
| CN114878707B (en) * | 2022-04-27 | 2023-11-07 | 湖南省药品检验检测研究院 | High performance liquid chromatography for determining N-nitrosodimethylamine in ranitidine and solid preparation thereof |
| CN115389673B (en) * | 2022-09-01 | 2024-05-14 | 湖南省药品检验检测研究院 | Method for determining genotoxic impurities in ranitidine hydrochloride by liquid chromatography-mass spectrometry |
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