CN114354780B - Method for detecting impurity content in ammonia bromine terro oral solution - Google Patents
Method for detecting impurity content in ammonia bromine terro oral solution Download PDFInfo
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- 239000012535 impurity Substances 0.000 title claims abstract description 93
- 238000000034 method Methods 0.000 title claims abstract description 29
- 229940100688 oral solution Drugs 0.000 title claims abstract description 29
- HHAGWXCTPQVPJV-UHFFFAOYSA-N N.[Br] Chemical compound N.[Br] HHAGWXCTPQVPJV-UHFFFAOYSA-N 0.000 title abstract description 7
- 239000000243 solution Substances 0.000 claims abstract description 36
- 238000001514 detection method Methods 0.000 claims abstract description 20
- 229960005174 ambroxol Drugs 0.000 claims abstract description 18
- JBDGDEWWOUBZPM-XYPYZODXSA-N ambroxol Chemical compound NC1=C(Br)C=C(Br)C=C1CN[C@@H]1CC[C@@H](O)CC1 JBDGDEWWOUBZPM-XYPYZODXSA-N 0.000 claims abstract description 17
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 12
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims abstract description 10
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 8
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims abstract description 7
- 235000019838 diammonium phosphate Nutrition 0.000 claims abstract description 7
- 239000007864 aqueous solution Substances 0.000 claims abstract description 6
- 238000010828 elution Methods 0.000 claims abstract description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000000945 filler Substances 0.000 claims abstract description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims abstract description 4
- 239000000377 silicon dioxide Substances 0.000 claims abstract description 4
- 238000012937 correction Methods 0.000 claims description 12
- 238000002347 injection Methods 0.000 claims description 6
- 239000007924 injection Substances 0.000 claims description 6
- RCPAZWISSAVDEA-UHFFFAOYSA-N 2-amino-3,5-dibromobenzaldehyde Chemical compound NC1=C(Br)C=C(Br)C=C1C=O RCPAZWISSAVDEA-UHFFFAOYSA-N 0.000 claims description 4
- AJOQVCHLLULVGK-UHFFFAOYSA-N 4-(6,8-dibromo-2,4-dihydro-1h-quinazolin-3-yl)cyclohexan-1-ol Chemical compound C1CC(O)CCC1N1CC2=CC(Br)=CC(Br)=C2NC1 AJOQVCHLLULVGK-UHFFFAOYSA-N 0.000 claims description 4
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 4
- AZFNTOYMFZMLCB-KYZUINATSA-N NC(C(Br)=CC(Br)=C1)=C1N[C@H](CC1)CC[C@@H]1O Chemical compound NC(C(Br)=CC(Br)=C1)=C1N[C@H](CC1)CC[C@@H]1O AZFNTOYMFZMLCB-KYZUINATSA-N 0.000 claims description 4
- PACIGGABLPTEDO-XYPYZODXSA-N NC1=C(C=N[C@@H]2CC[C@H](CC2)O)C=C(C=C1Br)Br Chemical compound NC1=C(C=N[C@@H]2CC[C@H](CC2)O)C=C(C=C1Br)Br PACIGGABLPTEDO-XYPYZODXSA-N 0.000 claims description 4
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 4
- GHUMSGGCKVMYGH-UHFFFAOYSA-N (2-amino-3,5-dibromophenyl)methanol Chemical compound NC1=C(Br)C=C(Br)C=C1CO GHUMSGGCKVMYGH-UHFFFAOYSA-N 0.000 claims 2
- 239000002904 solvent Substances 0.000 abstract description 23
- 239000000463 material Substances 0.000 abstract description 12
- 239000012452 mother liquor Substances 0.000 description 60
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 42
- QNVKOSLOVOTXKF-UHFFFAOYSA-N 4-[(2-amino-3,5-dibromophenyl)methylamino]cyclohexan-1-ol;hydron;chloride Chemical compound Cl.NC1=C(Br)C=C(Br)C=C1CNC1CCC(O)CC1 QNVKOSLOVOTXKF-UHFFFAOYSA-N 0.000 description 26
- 229960000985 ambroxol hydrochloride Drugs 0.000 description 26
- 229960001399 clenbuterol hydrochloride Drugs 0.000 description 20
- OPXKTCUYRHXSBK-UHFFFAOYSA-N clenbuterol hydrochloride Chemical compound Cl.CC(C)(C)NCC(O)C1=CC(Cl)=C(N)C(Cl)=C1 OPXKTCUYRHXSBK-UHFFFAOYSA-N 0.000 description 20
- 239000013558 reference substance Substances 0.000 description 20
- 238000005303 weighing Methods 0.000 description 19
- XUKUURHRXDUEBC-SXOMAYOGSA-N (3s,5r)-7-[2-(4-fluorophenyl)-3-phenyl-4-(phenylcarbamoyl)-5-propan-2-ylpyrrol-1-yl]-3,5-dihydroxyheptanoic acid Chemical compound C=1C=CC=CC=1C1=C(C=2C=CC(F)=CC=2)N(CC[C@@H](O)C[C@H](O)CC(O)=O)C(C(C)C)=C1C(=O)NC1=CC=CC=C1 XUKUURHRXDUEBC-SXOMAYOGSA-N 0.000 description 16
- AAEQXEDPVFIFDK-UHFFFAOYSA-N 3-(4-fluorobenzoyl)-2-(2-methylpropanoyl)-n,3-diphenyloxirane-2-carboxamide Chemical compound C=1C=CC=CC=1NC(=O)C1(C(=O)C(C)C)OC1(C=1C=CC=CC=1)C(=O)C1=CC=C(F)C=C1 AAEQXEDPVFIFDK-UHFFFAOYSA-N 0.000 description 16
- 238000012360 testing method Methods 0.000 description 12
- 239000000126 substance Substances 0.000 description 9
- 238000011084 recovery Methods 0.000 description 8
- 239000000523 sample Substances 0.000 description 8
- 239000000047 product Substances 0.000 description 7
- 238000007865 diluting Methods 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 238000005259 measurement Methods 0.000 description 6
- 239000012085 test solution Substances 0.000 description 6
- 230000004807 localization Effects 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 229940079593 drug Drugs 0.000 description 4
- 238000003255 drug test Methods 0.000 description 4
- 238000012417 linear regression Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 210000003097 mucus Anatomy 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- -1 ambroxol compound Chemical class 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
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- 239000012488 sample solution Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000005696 Diammonium phosphate Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 229940015596 ambroxol oral solution Drugs 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 239000012491 analyte Substances 0.000 description 1
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- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a method for detecting the content of impurities in an ammonia bromine Terra oral solution, which adopts high performance liquid chromatography to detect, takes octadecylsilane chemically bonded silica as a filling agent, and takes a 9-11 mol/L diammonium hydrogen phosphate aqueous solution as a mobile phase A; and (3) performing gradient elution by using an acetonitrile solution containing 0.1vt.% of tetrahydrofuran as a mobile phase B, wherein the column temperature is 35-45 ℃, and the flow rate is 0.8-1.2 ml per minute. The method for detecting the impurity content in the oral solution of the ambroxol effectively monitors the quality of the oral solution of the ambroxol, has high specificity, and does not interfere with the detection of known impurities by blank auxiliary materials and solvents.
Description
Technical Field
The invention belongs to the field of detection methods, relates to a detection method of related substances in an oral solution of amoebrolo, and particularly relates to detection of organic impurities such as degradation products of the oral solution of amoebrolo.
Background
The ambroxol oral solution is a compound preparation consisting of ambroxol hydrochloride and clenbuterol hydrochloride, wherein the ambroxol hydrochloride is a mucus dissolving agent, has the characteristics of mucus removal promotion and secretion dissolution, and can promote the removal of viscous secretions in respiratory tracts and reduce the retention of mucus; the clenbuterol hydrochloride is a beta 2 receptor agonist, belongs to an epinephrine-like medicament, can relax bronchial smooth muscle and has an asthma relieving effect.
At present, the related substance detection method of the ammonia bromine Terra oral solution is not seen in pharmacopoeia of various countries. The national medicine standard WS1- (X-118) -2006Z of the oral solution of ambroxol only has a content determination method of each component, and related substances of the product are not controlled by the quality standard.
The detection method of related substances in the quality standard of ambroxol hydrochloride, which is collected in pharmacopoeia of various countries, is only suitable for single preparation of ambroxol hydrochloride, and cannot effectively separate the components of clenbuterol hydrochloride and preservative contained in the product.
Chinese patent CN113049687A discloses a method for detecting related substances of ambroxol hydrochloride raw materials and injection, which adopts isocratic elution mode, because the product is an ambroxol compound preparation and an oral solution, contains various auxiliary materials, the interference generated by the method is far greater than that of the raw material medicines and the injection, the method can not effectively separate the auxiliary materials and clenbuterol hydrochloride contained in the product, and can not realize the purpose of rapidly detecting trace impurities.
In summary, there is no detection method for related substances in the oral solution of ambroxol, and the impurity condition in the oral solution of ambroxol cannot be effectively monitored, so as to ensure the quality of the oral solution of ambroxol, and an effective method suitable for detecting the impurities of the variety is urgently needed to be developed.
Disclosure of Invention
The invention aims to: in order to effectively control the content of impurities in the oral solution of the ammonia bromine and further improve the product quality of the oral solution of the ammonia bromine, the invention provides a method for detecting related substances of the oral solution of the ammonia bromine. The method of the invention can eliminate the interference of auxiliary materials and solvent peaks when detecting related substances in the oral solution of the ambroxol by adopting high performance liquid chromatography, and the separation degree between each known impurity and a main component chromatographic peak meets the requirement.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for detecting the content of impurities in an oral solution of ambroxol adopts high performance liquid chromatography for detection, octadecylsilane chemically bonded silica is used as a filling agent, and an aqueous solution of 9-11 mol/L diammonium hydrogen phosphate is used as a mobile phase A; and (3) performing gradient elution by using an acetonitrile solution containing 0.1vt.% of tetrahydrofuran as a mobile phase B, wherein the column temperature is 35-45 ℃, and the flow rate is 0.8-1.2 ml per minute.
Specifically, the pH of the aqueous solution of diammonium hydrogen phosphate is adjusted to 8.0-8.4 by ammonia water.
Specifically, the gradient elution procedure is:
| time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0 | 70 | 30 |
| 5 | 70 | 30 |
| 30 | 45 | 45 |
| 45 | 45 | 70 |
| 46 | 70 | 30 |
| 60 | 70 | 30 |
Specifically, the impurities include any one or more of 2-amino-3, 5-dibromomethanol, trans-4- (6, 8-dibromo-1, 4-dihydroquinazolin-3 (2H) -yl) cyclohexanol, trans-4- [ [ (2-amino-3, 5-dibromophenyl) methylene ] amino ] cyclohexanol, trans-4- [ (2-amino-3, 5-dibromophenyl) amino ] cyclohexanol, and 2-amino-3, 5-dibromobenzaldehyde.
Preferably, the high performance liquid chromatography uses an ultraviolet detection wavelength of 248nm, the column temperature is 40 ℃, and the flow rate is 1.0ml per minute.
Preferably, the injection temperature is 6 ℃.
Preferably, the aqueous solution of diammonium phosphate is adjusted to pH 8.2 with ammonia.
Preferably, the correction factor for 2-amino-3, 5-dibromomethanol is 0.70, the correction factor for trans-4- (6, 8-dibromo-1, 4-dihydroquinazolin-3 (2H) -yl) cyclohexanol is 1.62, the correction factor for trans-4- [ [ (2-amino-3, 5-dibromophenyl) methylene ] amino ] cyclohexanol is 0.86, and the correction factor for trans-4- [ (2-amino-3, 5-dibromophenyl) amino ] cyclohexanol is 1.17, and the correction factor for 2-amino-3, 5-dibromobenzaldehyde is 0.76.
The invention relates to a method for detecting the content of impurities in an oral solution of ambroxol, which comprises the following steps:
a. taking about 4.5g of oral solution of ambroxol, precisely weighing, placing in a 10ml measuring flask, adding a proper amount of solvent (mobile phase A-mobile phase B (70:30)), shaking uniformly, diluting with the solvent to scale, shaking uniformly, filtering, and taking the subsequent filtrate as a test solution.
b. Taking about 4.5g of blank auxiliary material solution, accurately weighing, placing in a 10ml measuring flask, adding a proper amount of solvent, shaking up, diluting with solvent to scale, shaking up, filtering, and taking the subsequent filtrate as blank auxiliary material determination solution.
c. Preparation of control solution
Clenbuterol hydrochloride mother liquor: taking a proper amount of clenbuterol hydrochloride reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of clenbuterol hydrochloride per 1 ml.
Ambroxol hydrochloride mother liquor: taking a proper amount of ambroxol hydrochloride reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of ambroxol hydrochloride per 1 ml.
Table 1 is a table of each impurity information:
TABLE 1
Impurity A mother liquor: taking a proper amount of the reference substance containing the impurity A, accurately weighing, and adding methanol to dissolve to prepare solution containing 50 mu g of the impurity A in each 1 ml.
Mother liquor of impurity B: taking a proper amount of the reference substance of the impurity B, precisely weighing, and adding methanol to dissolve to prepare solution containing 50 mu g of the impurity B in each 1 ml.
Mother liquor of impurity C: taking a proper amount of impurity C reference substance, precisely weighing, and adding methanol to dissolve to obtain a solution containing impurity C50 μ g per 1 ml.
Mother liquor of impurity D: taking a proper amount of the impurity D reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of the impurity D per 1 ml.
Mother liquor of impurity E: taking a proper amount of reference substance containing the impurity E, accurately weighing, and adding methanol to dissolve to obtain a solution containing 50 μ g of the impurity E per 1 ml.
And (3) mixing a solution I: respectively taking 2ml of ambroxol hydrochloride mother liquor, impurity A mother liquor, impurity B mother liquor, impurity C mother liquor, impurity D mother liquor and impurity E mother liquor, putting the ambroxol hydrochloride mother liquor, the impurity A mother liquor, the impurity B mother liquor, the impurity C mother liquor, the impurity D mother liquor and the impurity E mother liquor into a 10ml measuring flask, adding a solvent to dilute to a scale, and shaking up.
Clenbuterol hydrochloride positioning solution: taking 0.2ml of clenbuterol hydrochloride mother liquor, placing the clenbuterol hydrochloride mother liquor into a 10ml measuring flask, adding a solvent to dilute the clenbuterol hydrochloride mother liquor to a scale mark, and shaking up.
Has the beneficial effects that:
the method for detecting the impurity content in the oral solution of the ambroxol effectively monitors the quality of the oral solution of the ambroxol, has high specificity, and does not interfere with the detection of known impurities by blank auxiliary materials and solvents. By optimally combining different chromatographic conditions and adding 0.1vt.% of tetrahydrofuran, the serious interference generated by an auxiliary material solution in the detection process is eliminated, the separation effect between each impurity and a main component of the product is effectively improved, and the accurate and rapid quantitative detection of trace impurities in a complex system is realized. The mobile phase adopts high pH value (pH is 8.2) and low temperature injection at 6 ℃, so that the amount of impurities in the sample solution is basically unchanged within 12 hours, and new preparation is not needed, thus more efficient batch detection is possible.
Drawings
The foregoing and/or other advantages of the invention will become more apparent from the following detailed description of the invention when taken in conjunction with the accompanying drawings.
FIG. 1 is a HPLC (high Performance liquid chromatography) chart of the impurities and blank auxiliary materials detected in the oral solution of the invention.
Detailed Description
The invention will be better understood from the following examples.
The reagents used in the examples of the present invention were all chromatographically pure, unless otherwise indicated.
1. Reagents and materials
Diamine hydrogen phosphate, ammonia water, phosphoric acid, acetonitrile and tetrahydrofuran.
Ambroxol hydrochloride reference substance (source: China institute for testing food and drug, batch number: 100599-202106, content: 100%);
a clenbuterol hydrochloride reference substance (source: China institute for food and drug assay, batch number: 100072-201503, content: 90.5%);
impurity A as a reference (the source is Chinese food and drug testing institute, batch No. 510025-201702, the content is 100%);
impurity B as a reference (the source: China institute for testing and testing food and drug; batch No. 510026-202003, content: 100%);
impurity C as a reference (the source is China institute for food and drug assay, lot No. 510027-201902, content: 100%);
impurity D as a control (the source is China institute for testing food and drug, lot number: 510028-201902, content: 100%);
impurity E as a reference (the source: China institute for food and drug assay; batch No. 510029-202003, content: 99.9%);
blank auxiliary material (from: self-grinding, batch number: F-211103)
Oral solution of ambroxol (from: self-research, batch No. 211103).
2. Main instrument
Thermo U3000 high performance liquid chromatograph, sidoris BT125D electronic balance, mettler FE-28pH meter.
3. Chromatographic conditions
And (3) chromatographic column: octadecylsilane chemically bonded silica was used as a filler (Gemini 5 μm NX-C18110A, 4.6X 250mm)
Flow rate: 1.0ml/min
Column temperature: 40 deg.C
Wavelength: 248nm
Sample injection amount: 20 μ l
Sample injector temperature: 6 deg.C
4. Method establishment
TABLE 2 mobile phase and gradient elution program optimization screening
Finally, the mobile phase of serial number 5 and the gradient elution program are selected as the related substances of the product for detection.
5. Methodology validation
5.1 specificity
Clenbuterol hydrochloride mother liquor: taking a proper amount of clenbuterol hydrochloride reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of clenbuterol hydrochloride per 1 ml.
Ambroxol hydrochloride mother liquor: taking a proper amount of ambroxol hydrochloride reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of ambroxol hydrochloride per 1 ml.
Mother liquor of impurity A: taking a proper amount of the reference substance of the impurity A, precisely weighing, and adding methanol to dissolve to prepare solution containing 50 mu g of the impurity A in each 1 ml.
Mother liquor of impurity B: taking a proper amount of reference substance containing the impurity B, accurately weighing, and adding methanol to dissolve to obtain a solution containing 50 μ g of the impurity B per 1 ml.
Mother liquor of impurity C: taking a proper amount of impurity C reference substance, precisely weighing, and dissolving with methanol to obtain solution containing impurity C50 μ g per 1 ml.
Mother liquor of impurity D: taking a proper amount of the impurity D reference substance, precisely weighing, and adding methanol to dissolve to prepare a solution containing 50 mu g of the impurity D per 1 ml.
Mother liquor of impurity E: taking a proper amount of reference substance containing the impurity E, accurately weighing, and adding methanol to dissolve to obtain a solution containing 50 μ g of the impurity E per 1 ml.
Ambroxol hydrochloride positioning solution: taking 2ml of ambroxol hydrochloride mother liquor, placing the ambroxol hydrochloride mother liquor into a 10ml measuring flask, adding a solvent to dilute the ambroxol hydrochloride mother liquor to a scale, and shaking the ambroxol hydrochloride mother liquor uniformly.
Impurity a localization solution: and (3) taking 2ml of the mother liquor of the impurity A, putting the mother liquor into a 10ml measuring flask, adding a solvent to dilute the mother liquor to a scale, and shaking up the mixture.
Impurity B localization solution: and taking 2ml of the mother liquor of the impurity B, placing the mother liquor into a 10ml measuring flask, adding a solvent to dilute the mother liquor to a scale, and shaking the mother liquor uniformly.
Impurity C localization solution: and (3) taking 2ml of the mother liquor of the impurity C, putting the mother liquor of the impurity C into a 10ml measuring flask, adding a solvent to dilute the mother liquor to a scale, and shaking the mother liquor uniformly.
Impurity D localization solution: and (3) taking 2ml of the mother liquor of the impurity D, putting the mother liquor into a 10ml measuring flask, adding a solvent to dilute the mother liquor to a scale, and shaking up the mixture.
Impurity E localization solution: taking 2ml of the impurity E mother liquor, placing the impurity E mother liquor into a 10ml measuring flask, adding a solvent to dilute the impurity E mother liquor to a scale, and shaking up.
And (3) mixing liquid I: respectively taking 2ml of ambroxol hydrochloride mother liquor, impurity A mother liquor, impurity B mother liquor, impurity C mother liquor, impurity D mother liquor and impurity E mother liquor, placing the two solutions into a 10ml measuring flask, adding a solvent to dilute the solutions to a scale, and shaking up the solutions.
Clenbuterol hydrochloride positioning solution: taking 0.2ml of clenbuterol hydrochloride mother liquor, placing the clenbuterol hydrochloride mother liquor in a 10ml measuring flask, adding a solvent to dilute the clenbuterol hydrochloride mother liquor to a scale, and shaking up the clenbuterol hydrochloride mother liquor.
Blank adjuvant solution: taking about 4.5g of blank auxiliary materials, precisely weighing, placing in a 10ml measuring flask, adding a proper amount of solvent, shaking up, diluting with the solvent to a scale, and shaking up.
TABLE 3 specialization
5.2 Linear and Range
The linear relationship is plotted as a function of measured response signal (peak area) versus analyte concentration, and a linear regression is performed using the least squares method, requiring the linear regression coefficient R 2 Should be no less than 0.990.
For unknown impurities, the linearity and range of a single unknown impurity were examined instead with the ambroxol hydrochloride control.
TABLE 4 Linear measurement of ambroxol hydrochloride
TABLE 5 Linear measurement of impurity A
TABLE 6 Linear measurement of impurity B
TABLE 7 Linear measurement of impurity C
TABLE 8 Linear measurement of impurity D
TABLE 9 Linear measurement of impurity E
5.3 known impurity correction factor
The calibration factor of the known impurity is determined by standard curve method, that is, the impurity reference substance and the main component reference substance are precisely weighed, the series of solutions (covering report limit and standard limit) are prepared respectively and injected respectively, linear regression is carried out on the response factor (peak area) by the concentration of the sample according to the least square method, two standard curves are obtained, and the ratio of the main component to the known impurity slope is the calibration factor.
| Name(s) | Slope of | Correction factor |
| Ambroxol hydrochloride | 0.5149 | / |
| Impurity A | 0.7332 | 0.70 |
| Impurity B | 0.3186 | 1.62 |
| Impurity C | 0.5984 | 0.86 |
| Impurity D | 0.439 | 1.17 |
| Impurity E | 0.6767 | 0.76 |
5.4 detection and quantitation limits
For known impurities, the limit of detection (LOD) and limit of quantitation (LOQ) are determined according to the signal-to-noise ratio method. A stock solution of an impurity of known concentration is diluted to a low concentration sample and the measured signal is compared to the signal at the blank (baseline noise) to calculate the lowest concentration or percentage that can be reliably detected, both the limit of detection and the limit of quantitation should be below the reported limit (0.05%).
For unknown impurities, the detection limit and the quantification limit of a single unknown impurity are inspected by using an ambroxol hydrochloride reference substance instead.
Diluting the mixed solution of each impurity and the ambroxol hydrochloride reference substance step by step, determining LOQ according to the signal-to-noise ratio S/N being more than or equal to 10, and determining LOD according to the S/N being more than or equal to 3.
TABLE 10 results on substance-quantitation limits
| Sample (I) | Quantitative limit concentration (mu g/ml) | Relative sample concentration (%) | Signal-to-noise ratio (S/N) |
| Ambroxol hydrochloride | 0.076 | 0.013 | 14.5 |
| Impurity A | 0.022 | 0.004 | 17.3 |
| Impurity B | 0.072 | 0.012 | 16.1 |
| Impurity C | 0.049 | 0.008 | 15.1 |
| Impurity D | 0.036 | 0.006 | 12.4 |
| Impurities in the productE | 0.039 | 0.007 | 13.7 |
TABLE 11 related substances-detection Limit results
5.5 accuracy test
Accuracy is determined by the recovery of each known impurity at three different concentrations, 80%, 100% and 120% of the limit of the impurity added to the test article. The accuracy of the known impurities is determined by adding a known amount of impurities and determining the ratio (recovery) between the test result and the theoretical value of the known impurities in the sample to be added, expressed in percent, with the requirement that the recovery of each sample be between 95.0% and 105.0% and the RSD be less than or equal to 5%.
TABLE 12 recovery of impurity A
TABLE 13 recovery of impurity B
TABLE 14 recovery of impurity C
TABLE 15 recovery of impurity D
TABLE 16 recovery of impurity E
5.5 repeatability test
The repeatability test is carried out by preparing 6 sample solutions and testing, and the absolute deviation of the impurity variation of 6 times of data results is required to be not more than 0.1%.
Test solution: taking about 4.5g of oral solution of ambroxol, accurately weighing, placing in a 10ml measuring flask, adding a proper amount of solvent (mobile phase A-mobile phase B (70:30)), shaking uniformly, diluting to scale with solvent, and shaking uniformly to obtain the final product. 6 portions of the mixture are prepared by the same method.
TABLE 17 materials-repeatability test results (area normalization)
5.6 stability of test solution
Test solution: taking about 4.5g of oral solution of ambroxol, precisely weighing, placing in a 10ml measuring flask, adding a proper amount of solvent (mobile phase A-mobile phase B (70:30)), shaking uniformly, diluting with the solvent to scale, and shaking uniformly to obtain the final product.
And (3) placing the test solution at the low temperature of 6 ℃, injecting samples at different time points respectively for analysis, and recording a chromatogram map.
TABLE 18 test results of stability of test solutions (area normalization method)
The invention provides a method and a concept for detecting the content of impurities in ammonia bromine terro oral solution, and a method and a way for implementing the technical scheme are many, the above description is only a preferred embodiment of the invention, and it should be noted that, for those skilled in the art, a plurality of improvements and modifications can be made without departing from the principle of the invention, and the improvements and modifications should also be regarded as the protection scope of the invention. All the components not specified in the present embodiment can be realized by the prior art.
Claims (3)
1. A method for detecting the content of impurities in an oral solution of ambroxol is characterized in that high performance liquid chromatography is adopted for detection, octadecylsilane chemically bonded silica is used as a filling agent, and an aqueous solution of 9-11 mol/L diammonium hydrogen phosphate is used as a mobile phase A; performing gradient elution by taking an acetonitrile solution containing 0.1vt.% tetrahydrofuran as a mobile phase B, wherein the column temperature is 35-45 ℃, and the flow rate is 0.8-1.2 ml per minute;
adjusting the pH value of the diammonium hydrogen phosphate aqueous solution to 8.0-8.4 by using ammonia water;
the impurities include 2-amino-3, 5-dibromobenzyl alcohol, trans-4- (6, 8-dibromo-1, 4-dihydroquinazolin-3 (2H) -yl) cyclohexanol, trans-4- [ [ (2-amino-3, 5-dibromophenyl) methylene ] amino ] cyclohexanol, trans-4- [ (2-amino-3, 5-dibromophenyl) amino ] cyclohexanol, and 2-amino-3, 5-dibromobenzaldehyde;
the high performance liquid chromatography uses ultraviolet with a wavelength of 248nm for detection, the column temperature is 40 ℃, and the flow rate is 1.0ml per minute;
the sample injection temperature is 6 ℃;
the gradient elution procedure was:
2. The method of claim 1, wherein the ammonium phosphate dibasic solution is adjusted to a pH of 8.2 with ammonia water.
3. The method for detecting the content of impurities in an oral solution of amobromotetrol according to claim 1, wherein the correction factor for 2-amino-3, 5-dibromobenzyl alcohol is 0.70, the correction factor for trans-4- (6, 8-dibromo-1, 4-dihydroquinazolin-3 (2H) -yl) cyclohexanol is 1.62, the correction factor for trans-4- [ [ (2-amino-3, 5-dibromophenyl) methylene ] amino ] cyclohexanol is 0.86, and the correction factor for trans-4- [ (2-amino-3, 5-dibromophenyl) amino ] cyclohexanol is 1.17, and the correction factor for 2-amino-3, 5-dibromobenzaldehyde is 0.76.
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| CN113049687A (en) * | 2019-12-27 | 2021-06-29 | 石药集团欧意药业有限公司 | Method for detecting related substances of ambroxol hydrochloride raw material and injection |
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