CN109799292B - Mesalazine enema related substance determination method - Google Patents
Mesalazine enema related substance determination method Download PDFInfo
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Abstract
The invention provides a method for measuring related substances of mesalazine enema and discloses a method for analyzing related substances of mesalazine enema. The method adopts high performance liquid chromatography, uses octadecylsilane chemically bonded silica as chromatographic column filler, adopts an ultraviolet detector, selects proper organic phase, buffer solution and water as mobile phase, and adopts gradient elution, thereby effectively solving the separation problem of mesalazine and impurities. The method has strong specificity and good repeatability, and can ensure the controllability of the process and the degradation impurities in the mesalazine.
Description
Technical Field
The invention relates to the technical field of determination of related substances of medicines, in particular to a method for determining related substances of a mesalazine enema.
Background
Ulcerative colitis is a disease existing all over the world, in recent years, the incidence rate of ulcerative colitis tends to increase in China, and sulfasalazine (SASP) is mainly used for clinical treatment, although the ulcerative colitis has a certain curative effect, the incidence rate of side effects is high, and the ulcerative colitis cannot be applied to patients with sulfanilamide allergy. Mesalazine was first developed and marketed by Hokka pharmaceutical factory in Germany, and compared with SASP, its main chemical structure is that 5-aminosalicylic acid active ingredient is retained, and sulfapyridine group is removed, so that it overcomes the defect of SASP, and is a new medicine for treating ulcerative colitis, and has been extensively used in Western countries. At present, due to the influence of a plurality of factors such as the deterioration of living environment and living habits of people, the incidence rate of ulcerative colitis is increased year by year, no better medicine is used for treating the ulcerative colitis before the emergence of mesalazine, and the clinical popularization is continuously realized with the advantages of good curative effect, few adverse reactions and the like after the emergence of the mesalazine.
The main pharmacological actions of mesalazine are to directly inhibit colonic peroxidase and inhibit the synthesis and release of inflammatory mediators including prostaglandin, leukotriene and the like; inhibiting the activity of platelet activating factor; free radical scavenging, intestinal mucosa injury and irritation reducing, SASP side effect avoiding, and is suitable for patients with SASP and glucocorticoid ineffectiveness or intolerance. The effective rate is obviously higher than SASP from the symptom relief and colonoscope change. In addition, the level of TNF-a and IL-8 in the serum of a patient can be obviously reduced, which is probably one of the action mechanisms of the medicine for treating ulcerative colitis.
The anti-inflammatory mechanism of action of mesalazine is not fully understood. In vitro studies show that mesalazine has certain influence on the content of intestinal mucosa prostaglandin, has the function of clearing active oxygen free radicals, and may play a certain role in inhibiting lipoxygenase. After rectal administration of mesalazine enema, mesalazine acts mainly locally on the intestinal mucosa and submucosal tissues.
The related substances are mainly starting materials, intermediates, polymers, side reaction products brought in during the production process, degradation products in the storage process and the like. The related substances are closely related to the quality, safety and curative effect of the medicine, and the existence of the related substances may reduce the curative effect of the medicine and even cause toxic and side effects, so the types and the contents of the related substances in the medicine must be controlled by a proper detection and analysis method to ensure the quality of the medicine.
The detection method for collecting and carrying related substances of mesalazine in USP (USP), BP (back propagation) standards and import quality labels at present is found through research on the detection method: firstly, the USP standard adopts isocratic elution, and can not completely separate each impurity in mixed impurities; secondly, the interference of gradient peaks at the positions of a main peak and an impurity peak in the BP standard is obvious, the impurities A and C independently adopt another chromatographic condition, and perchloric acid is added into a mobile phase in the chromatographic condition for detecting the impurities A and C, so that a chromatographic column is easily damaged; and thirdly, the phenomenon of incomplete detection of impurities exists in the imported quality standard, and the separation degree of partial impurities is not high, so that accurate detection cannot be realized. The method can detect the related substances of the mesalazine under the same chromatographic condition, can effectively separate impurities, and has important significance for quality control.
Disclosure of Invention
In order to solve the technical problems, the invention provides the mesalazine enema liquid which has good specificity and separation degree, can effectively separate possible degradation impurities and process impurities from main components, has high sensitivity and can be applied to the quality control of the mesalazine enema liquid.
The inventor identifies 11 impurities aiming at the synthesis process and the degradation condition of mesalazine.
The inventor screens under conditions such as gradient elution procedures and the like, takes the requirements of 11 related substances into consideration, finally establishes a proper related substance analysis method, enables the baseline separation between the main component and each related substance and between each related substance to be achieved, and achieves the aim of the invention.
The embodiment of the invention firstly provides a method for measuring related substances of mesalazine, which adopts a high performance liquid chromatography for measurement, wherein the chromatographic conditions of the high performance liquid chromatography comprise:
octadecylsilane chemically bonded silica is used as a filler for the chromatographic column;
dissolving 4.35g of dipotassium phosphate in water to 1000ml, adjusting the pH to 7.2) -TBAH solution with phosphoric acid (80 ml of 10% tetrabutylammonium hydroxide aqueous solution, diluting with water to 200ml, adjusting the pH to 7.5 with sulfuric acid solution (1 → 2) and methanol-water (250: 50: 100: 600) as mobile phase A, dissolving 4.35g of dipotassium phosphate in water to 1000ml, adjusting the pH to 7.2) -TBAH solution with phosphoric acid (80 ml of 10% tetrabutylammonium hydroxide aqueous solution, diluting with water to 200ml and adjusting the pH to 7.5 with sulfuric acid solution (1 → 2) and methanol-water (250: 50: 500: 200) as mobile phase B;
the gradient elution procedure was:
the column temperature was 40 ℃; the detection wavelength is 230 nm; the flow rate was 1.0 mL/min.
Taking a proper amount of the product, precisely weighing, adding 0.01mol/L hydrochloric acid for dissolving, and diluting to prepare a solution containing about 1.0mg of mesalazine in each 1ml, wherein the solution is used as a test solution. Precisely measuring a proper amount of a test solution, adding 0.01mol/L hydrochloric acid, and diluting to obtain a solution containing 1 mu g of mesalazine in each 1ml, wherein the solution is used as a control solution.
Precisely weighing appropriate amount of impurity control substances, respectively, adding 0.01mol/L hydrochloric acid for dissolving, and quantitatively diluting to obtain mixed solution containing about 1 μ g of each impurity in 1ml as impurity control substance solution.
Precisely weighing appropriate amount of impurity reference substance and mesalazine reference substance respectively, adding 0.01mol/L hydrochloric acid for dissolving, and quantitatively diluting to obtain mixed solution containing about 1 μ g per 1ml, and using the mixed solution as system applicability solution.
The sample volume of the test solution, the control solution and the impurity control solution is 10 mul.
Drawings
FIG. 1 is a chromatogram of an elution procedure 1 in optimization of a method for determining related substances in mesalazine enema according to the present invention;
FIG. 2 is a chromatogram of an elution procedure 2 in optimization of a method for determining related substances in mesalazine enema according to the present invention;
FIG. 3 is an elution program 3 chromatogram (standard chromatogram in the present invention) in the optimization of the method for determining related substances in mesalazine enema according to the present invention;
FIG. 4 is a chromatogram of an elution procedure 4 in the optimization of the method for determining related substances in mesalazine enema according to the present invention;
FIG. 5 is a chromatogram of an elution procedure 5 in optimization of a method for determining related substances in mesalazine enema according to the present invention;
FIG. 6 is a chromatogram of USP standard in comparison with the method using the method for measuring substances related to mesalazine enema liquid of the present invention;
fig. 7 is a chromatogram of BP2017 standard in comparison with a method for determining related substances in mesalazine enema according to the present invention;
FIG. 8 is a chromatogram of imported registration standards in comparison with a method for determining substances related to mesalazine enema according to the present invention;
FIG. 9 is a chromatogram of a system suitability test solution determined using the method for determining substances related to mesalazine enema of the present invention;
FIG. 10 is a chromatogram of a special blank test solution determined by the method for determining substances related to mesalazine enema according to the present invention;
FIG. 11 is a chromatogram of a specific test solution determined by the method for determining substances related to mesalazine enema according to the present invention; the following are specifically mentioned: since the HPLC device used by the applicant is a web version, the workstation does not have any image editing and exporting functions, and can only be directly printed into a paper file, the drawing does not affect the interpretation of the chromatogram by the skilled person because of the inclination of the workstation and the printing device.
Detailed Description
The invention provides a method for measuring mesalazine related substances in order to measure the mesalazine related substances. In the technical scheme, the mesalazine is well separated from adjacent impurities and various impurities by optimizing the high performance liquid chromatography conditions. The technical solution of the present invention is specifically described below with reference to the accompanying drawings.
In the following embodiments, the drugs, reagents and instruments used are as follows:
TABLE 1 control information
| Numbering | Code | Name (R) | Batch number | Content (wt.) | Source |
| 1 | Impurity A | 4-aminophenols | L310R67 | 99.65％ | Bailingwei-medicine |
| 2 | Impurity C | 2-aminophenols | LB30R39 | 98.66％ | Bailingwei-medicine |
| 3 | Impurity E | 4-aminosalicylic acid | LGA0O08 | 98.78％ | Bailingwei-medicine |
| 4 | Impurity F | 3-aminosalicylic acid | L610P41 | 97.55％ | Bailingwei- |
| 5 | Impurity G | 2, 5-Dihydroxybenzoic acid | LDA0Q34 | 99.79％ | Bailingwei-medicine |
| 6 | Impurity H | Salicylic acid | LRC0Q52 | 100.74％ | Bailingwei-medicine |
| 7 | Impurity J | 3, 5-diaminosalicylic acid | WG016696150408001 | 95.0％ | Bide medicine |
| 8 | Impurity L | 2-Chlorobenzenecarboxylic acid | LA50Q22 | 98.97％ | Bailingwei-medicine |
| 9 | Impurity M | 2-chloro-5-nitrobenzoic acid | L270O88 | 99.96％ | Bailingwei- |
| 10 | Impurity N | 5-Nitrosalicylic acid | L370P28 | 99.52％ | Bailingwei-medicine |
| 11 | Impurity R | 3-Nitrosalicylic acid | NXZ6F | 99.4％ | TGI |
| 12 | —— | Mesalazine | 100565-200701 | 99.9％ | Inspection yard |
TABLE 2 sample information
TABLE 3 Instrument and Equipment information
| Name (R) | Model number | Manufacturer of the product |
| Electronic balance | BP211D | Sartorioμs |
| High performance liquid chromatograph | Agilent1200 | Agilent |
Preliminary optimization of chromatographic conditions
Under the preliminarily prepared quasi-chromatographic condition (elution procedure 1), the separation degree of each impurity is better, but the impurity N and the impurity R do not generate peaks under the retention time condition; the peak appearance condition of a single impurity can be known, the impurity N and the impurity R can appear in the next needle of sample, so under the condition that the change rate of the gradient flowing phase is not changed, the gradient time is properly prolonged for testing, and the elution program 1 is preliminarily optimized, wherein the elution program is optimized as follows: elution procedure 2 and elution procedure 3, and the maps are shown in figure 1, figure 2 and figure 3.
TABLE 4 elution procedure
TABLE 5 preliminary optimization results of chromatographic conditions
Under the condition of the high performance liquid chromatography, under the condition that the change rate of a gradient flowing phase ratio is not changed, known impurities can be completely detected by properly prolonging the gradient time, and the separation degrees of all the impurities except the impurity G and the impurity L are 1.05, are all better; thus the screening of the column is performed under the chromatographic conditions.
Further optimization of chromatographic conditions
On the basis of the elution procedure 3 method, the gradient elution procedure is adjusted, and the chromatographic conditions are further optimized so as to select better chromatographic conditions, wherein the elution procedure is optimized as follows: elution procedure 4 and elution procedure 5, and the maps are shown in figure 4 and figure 5.
TABLE 6 elution procedure
TABLE 7 results of varying elution procedures
In the high performance liquid chromatography conditions, under three elution procedures, the retention time of a main peak is basically consistent, the separation degree of the main peak and adjacent peaks is good, each impurity can be effectively detected, the detection capability of the main peak is consistent with that of a BP2017 standard impurity (11 known impurities with definite control limits in BP2017 raw materials and preparations can be effectively detected), and the separation degree between the impurity peaks is good.
The invention discloses a detection method of identified mesalazine enema related substances, wherein in the high performance liquid chromatography condition, the ratio of the used mobile phase A: phosphate buffer (ph7.2) -TBAH solution-methanol-water (250: 50: 100: 600); mobile phase B: phosphate buffer (ph7.2) -TBAH solution-methanol-water (250: 50: 500: 200) with gradient elution procedure as shown in table 8:
TABLE 8 elution procedure
| Time (min) | A | B％ | |
| 0 | 90 | 10 | |
| 60 | 15 | 85 | |
| 65 | 90 | 10 | |
| 75 | 90 | 10 |
The table shows the import drug registration standard JX20090090, EP9.0\ BP2017, USP40 and the invention standard for measuring substances related to mesalazine enema.
TABLE 9 relevant substance assay standards
The table shows the registration standard JX20090090, EP9.0\ BP2017 and the standard gradient elution procedure of the invention for determining related substances of mesalazine enema.
TABLE 10 gradient elution details
In the table, the maps of the import drug registration standard JX20090090, EP9.0\ BP2017, USP40 and the standard test result of the invention for measuring substances related to mesalazine enema liquid are shown in the attached figure 6, the attached figure 7 and the attached figure 8.
TABLE 11 measurement results of mixed impurities
TABLE 12 raw material measurement results
TABLE 13 results of measurements on samples
Comparing import drug registration standards JX20090090 and EP9.0\ BP2017 for determining related substances of mesalazine enema and the standard of the invention, the conclusion is as follows:
USP40 standard: with isocratic elution, it is not possible to separate the impurities, where impurity a overlaps with impurity J, and impurity M overlaps with the main peak; and the separation degree of each detected impurity is poor, the peaks of the impurities H and L are not separated, and the separation degree is less than 1.2.
BP2017 standard: the mixed solution of 11 impurities is adopted for sample injection, and all the impurities can be effectively detected under the chromatographic condition, but the main peak and the peak types of all the impurities are poor; the retention time of a main peak and an impurity peak is short, and the separation degree between the impurity peaks is poor; in the BP2017 standard, the impurities A and C independently adopt another chromatographic condition, and perchloric acid is added into a mobile phase for detecting the impurities A and C under the chromatographic condition, so that a chromatographic column is easily damaged.
③ import quality standard JX20090090 Standard: all impurities can be effectively detected, and all impurities can be effectively separated; however, the interference of the gradient peaks at the positions of the main peak and the impurity peak is obvious, and the impurity measurement is possibly interfered.
The standard of the invention is as follows: under the chromatographic condition, the separation degrees of all impurities and main peaks and adjacent peaks are good, and 11 impurities can be effectively detected; a known impurity F can be detected in a sample under USP40 chromatographic conditions, and the known impurity is not detected in the sample under the chromatographic conditions of import quality standard JX20090090 which can be interfered by a gradient peak; and 2 impurities can be detected in the sample under the BP2017 and the standard chromatographic conditions of the invention, wherein one of the impurities is the known impurity F.
Verifying content
1 system applicability
Preparing a mesalazine reference substance and a mixed solution of known impurities as a system applicability solution; precisely measuring 10 mu l of sample injection; the results are shown in Table 14 and the spectra are shown in FIG. 9.
TABLE 14 results of the System suitability measurement
The results show that: the main peak has better peak shape and symmetry, the theoretical plate number is high, the separation degree of the main peak and the adjacent impurity peak is better, and all impurities can be better separated; the analysis method is good in system applicability.
2 specialization
Blank auxiliary material interference test
Mixing diluent, blank auxiliary materials, raw materials, related substances, a reference (impurity A, C, E, F, G, H, J, N, R) and each solution of a sample, and precisely measuring 10 mu l of each solution for sample injection; the results are shown in tables 15 and 16. The map is shown in figure 10 and figure 11.
TABLE 15 results of the specificity test
The results show that: the diluent and the blank auxiliary materials do not interfere with the detection of main peaks and impurities.
TABLE 16 case of impurity separation
The results show that: the separation degree of mesalazine and adjacent impurities is more than 2.0, the separation degree of each impurity is more than 1.5, and the mesalazine and the adjacent impurities can be well separated.
② forced degradation test
Respectively preparing a mesalazine enema liquid storage solution and a blank auxiliary material storage solution, measuring the mesalazine enema liquid storage solution and the blank auxiliary material storage solution, respectively carrying out a forced degradation test according to the conditions in the table 17, finally measuring 10 mu l of a diluent, and injecting the diluent into a liquid chromatograph; the results are shown in tables 18 and 19.
TABLE 17 sample and blank adjuvant degradation conditions
Table 18 results of forced degradation of samples
Remarking: the test solution does not change color under the conditions of strong light, high temperature, oxidation and strong acid, and the test solution changes from colorless to light brown to brown under the condition of strong alkali.
TABLE 19 sample degradation impurity assignment analysis
The results show that: the degradation impurities are generated under the conditions of high temperature, strong light, strong acid, strong alkali and oxidation, the degradation degree of the main components is 95.25 to 99.96 percent, and the degradation condition of the impurities is basically consistent with that of the raw materials. Each degradation product is effectively separated from the main peak; the blank auxiliary materials do not generate degradation products and do not interfere the detection of sample impurities.
3 quantitative limit and detection limit
And (3) measuring the detection limit and the quantitative limit of each related substance of mesalazine by adopting a signal-to-noise ratio method. Respectively preparing stock solutions of various related substances of mesalazine, diluting to a certain concentration, injecting, calculating the ratio (signal-to-noise ratio) of peak height to noise, wherein the sample detection amount with the signal-to-noise ratio (S/N) of about 10 is the quantitative limit, the sample detection amount with the signal-to-noise ratio (S/N) of about 3 is the detection limit, and the result is shown in a table 20.
TABLE 20 test results for each impurity detection limit and quantitation limit
The results show that: the detection limit of each impurity is lower than the limit sample injection amount by 10ng, and the detection requirements of related substances are met; the method has high response to various related substances and can accurately control the content of the various related substances.
4 linear and range
Mesalazine and various impurity reference substances thereof are precisely weighed and dissolved and diluted into stock solution (concentration: about 2 mu g/ml) by adding 0.01mol/L hydrochloric acid. 6 parts of solutions with different concentrations are prepared and respectively injected, linear regression analysis is carried out by taking the sample injection amount as a horizontal coordinate and the peak area as a vertical coordinate, and a linear regression equation of mesalazine and each impurity thereof is obtained, and the result is shown in a table 21.
TABLE 21 results of the Linear test
| Name of impurity | Linear range | Linear equation of equations | Coefficient of linear correlation |
| Mesalazine | 3.10ng～60.64ng | y＝2.1453x+0.3722 | r＝0.999 |
| 4-aminophenols | 3.12ng～20.81ng | y＝3.6107x-0.7666 | r＝0.999 |
| 2-aminophenols | 2.85ng～19.00ng | y＝2.0311x-2.0495 | r＝0.997 |
| 4-aminobenzoic acid | 2.97ng～19.82ng | y＝3.2427x-1.2363 | r＝0.999 |
| 3-aminobenzoic acid | 3.18ng～21.20ng | y＝2.0251x+1.9682 | r＝0.999 |
| 2, 5-Dihydroxybenzoic acid | 3.03ng～20.22ng | y＝3.9321x-0.4948 | r＝0.999 |
| Salicylic acid | 3.05ng～20.30ng | y＝2.2391x+0.1946 | r＝0.999 |
| 3, 5-diaminosalicylic acid | 3.00ng～20.00ng | y＝2.8871x+1.1801 | r＝0.999 |
| 5-Nitrosalicylic acid | 2.97ng～19.80ng | y＝2.8417x-1.0882 | r＝0.999 |
| 3-Nitrosalicylic acid | 3.12ng～20.80ng | y＝2.5216x-0.8285 | r＝0.999 |
| 2-Chlorobenzenecarboxylic acid | 3.04ng～20.29ng | y＝2.7896x-1.7707 | r＝0.999 |
| 2-chloro-5-nitrobenzoic acid | 3.09ng～20.63ng | y＝2.3251x-0.885 | r＝0.999 |
The results show that: under the method, mesalazine and various impurities thereof can have good linear relation within a certain concentration range.
5 precision
And (3) repeatability test: and (3) adding a proper amount of each impurity reference substance into the self-prepared product solution to prepare 6 test sample solutions with impurity concentration of 0.1% of the labeled amount of mesalazine, injecting samples respectively, and calculating the content of each impurity. The results are shown in Table 22.
TABLE 22 results of the repeatability tests
The results show that: the RSD content of each impurity is less than 3 percent, which shows that the method has good repeatability.
6 recovery test
Preparing a control solution: taking a proper amount of each impurity reference substance, precisely weighing, adding 0.01mol/L hydrochloric acid to dissolve and dilute to prepare a mixed impurity stock solution with the concentration of 0.02mg/ml, precisely weighing 5ml of mixed impurity solution, placing the mixed impurity solution in a 100ml measuring flask, adding 0.01mol/L hydrochloric acid to dilute to scale, and preparing the mixed impurity reference solution with the concentration of 1 ug/ml.
Preparing a mesalazine self-prepared solution: precisely weighing a proper amount of the self-prepared mesalazine, and adding 0.01mol/L hydrochloric acid to dissolve and dilute the self-prepared mesalazine into a solution with the concentration of 15mg/ml for later use.
Preparing a sample solution: precisely measuring 10ml of the mixed impurity stock solution, putting the mixed impurity stock solution into a 100ml measuring flask, adding 0.01mol/L hydrochloric acid to dilute to scale, and preparing the mixed impurity stock solution (1) with the concentration of 2 mu g/ml; and precisely measuring 2.5ml of the mixed impurity stock solution, putting the mixed impurity stock solution into a 10ml measuring flask, and adding 0.01mol/L hydrochloric acid to dilute to scale to prepare the mixed impurity stock solution (2) with the concentration of 5 ug/ml. Precisely measuring 1ml of each stock solution (2), placing into 10ml measuring bottles, adding 150mg of the product, and adding 0.01mol/L hydrochloric acid to obtain 3 parts of a test solution with the labeled amount of mesalazine of 0.05%; and precisely measuring 3 parts of the stock solutions (1)5ml and 7.5ml respectively, placing the stock solutions in 10ml measuring bottles, adding 150mg of the prepared product respectively, and adding 0.01mol/L hydrochloric acid to prepare 3 parts of test solution with the mesalazine marked amount of 0.10% and the mesalazine marked amount of 0.15% respectively. The total of the above solutions was 9 parts, and the theoretical value and the measured value were compared to calculate the recovery rate. The results are shown in Table 23.
TABLE 23 recovery test results
The results show that: the average recovery rate of each impurity is 95.40-105.91%, and the RSD is less than 10%, which shows that the accuracy of the related substance determination method is good.
7 durability test
Durability tests were conducted by examining the column temperature variation. + -. 5 ℃, the relative variation in flow rate. + -. 20%, the pH variation of phosphate buffer. + -. 0.2, the flow phase variation. + -. 5%, the detection wavelength variation. + -. 5nm, and the column conditions, respectively. The mesalazine is taken from a product solution, the concentration of the mesalazine is 1mg/ml, and 10 mul is precisely measured and injected into a liquid chromatograph. The test results are shown in Table 24.
TABLE 24 results of different chromatographic conditions
The results show that: when all parameters are slightly changed, the theoretical plate numbers and the separation degrees of mesalazine and impurity peaks are not obviously changed, and the durability of the analysis method is good.
Claims (6)
1. A detection method of mesalazine related substances adopts high performance liquid chromatography, uses octadecylsilane chemically bonded silica as chromatographic column packing, adopts ultraviolet detector, and performs gradient elution; the method is characterized in that: the proportion of the mobile phase is as follows: phosphate buffer solution-tetrabutylammonium hydroxide solution-methanol-water volume ratio of 250: 50: 100: 600 as mobile phase A, and taking the volume ratio of phosphate buffer solution-tetrabutylammonium hydroxide solution-methanol-water as 250: 50: 500: 200 as a mobile phase B, wherein the phosphate buffer solution is as follows: taking 4.35g of dipotassium phosphate, adding water to dissolve the dipotassium phosphate to 1000ml, adjusting the pH value to 7.2 by using phosphoric acid, wherein the tetrabutyl ammonium hydroxide solution is as follows: taking 80ml of 10% tetrabutylammonium hydroxide aqueous solution, adding water to dilute the tetrabutylammonium hydroxide aqueous solution to 200ml, and adjusting the pH value to 7.5 by using sulfuric acid solution 1 → 2, wherein the volume percentage of the mobile phase A and the mobile phase B in the gradient elution procedure is as follows: 90:10 at 0 min; 15:85 at 60 min; 90:10 at 65 min; 90:10 at 75 min; the mesalazine related substances are 11 kinds as follows: 4-aminophenol; 2-aminophenol; 3-aminosalicylic acid; 2, 5-dihydroxybenzoic acid; salicylic acid; 5-nitrosalicylic acid; 4-aminosalicylic acid; 3, 5-diaminosalicylic acid; 3-nitrosalicylic acid; 2-chlorobenzoic acid; 2-chloro-5-nitrobenzoic acid.
2. The method for detecting a mesalazine-related substance according to claim 1, wherein: an ultraviolet detector is adopted, and the detection wavelength is 230 nm.
3. The method for detecting a mesalazine-related substance according to claim 1, wherein: the column temperature was 40 ℃.
4. The method for detecting a mesalazine-related substance according to claim 1, wherein: the flow rate was 1.0 mL/min.
5. The method for detecting a mesalazine-related substance according to claim 1, wherein: taking a proper amount of the product, precisely weighing, adding 0.01mol/L hydrochloric acid to dissolve and dilute the product to prepare a solution containing about 1.0mg of mesalazine in each 1ml as a test solution, precisely weighing a proper amount of the test solution, adding 0.01mol/L hydrochloric acid to dilute the test solution quantitatively to prepare a solution containing 1 mu g of mesalazine in each 1ml as a control solution.
6. The method for detecting a mesalazine-related substance according to claim 1, wherein: precisely weighing mesalazine related substance reference substances respectively, adding 0.01mol/L hydrochloric acid for dissolving, and quantitatively diluting to obtain mixed solution containing about 1 μ g of each mesalazine related substance per 1ml, wherein the mixed solution is used as the mesalazine related substance reference substance solution.
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| Determination of 5-aminosalicylic acid and related compounds in raw materials and pharmaceutical dosage forms by high-performance liquid chromatography;C. GANDINI et al;《Journal of Chromatography》;19911231;第540卷;第416-421页 * |
| Determination of Mesalamine Related Impurities from Drug Product by Reversed Phase Validated UPLC Method;TRIVEDI RAKSHIT KANUBHAI et al;《E-Journal of Chemistry》;20111231;第8卷(第1期);第131-148页 * |
| HPLC法测定美沙拉嗪中异亚丙基丙酮;董煜 等;《中国药品标准》;20171231;第18卷(第2期);第111页第2.1节；第112页第2.2.1-2.2.2节；图1 * |
| Ion-pairing high-performance liquid chromatographic method for the determination of Saminosalicylic acid and related impurities in bulk chemical;Brian S. Kersten et al;《Journal of Chromatography》;19911231;第588卷;第187-193页 * |
| 离子对色谱法测定颇得斯安灌肠剂中美沙拉嗦的含量及其降解产物;高青 等;《中国药科大学学报》;19991231;第30卷(第5期);第370-372页 * |
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