CN114264734B - Method for detecting related substances of aminolevulinic acid hydrochloride - Google Patents
Method for detecting related substances of aminolevulinic acid hydrochloride Download PDFInfo
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- ZGXJTSGNIOSYLO-UHFFFAOYSA-N 88755TAZ87 Chemical compound NCC(=O)CCC(O)=O ZGXJTSGNIOSYLO-UHFFFAOYSA-N 0.000 title claims abstract description 32
- 239000000126 substance Substances 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 24
- 229960000781 aminolevulinic acid hydrochloride Drugs 0.000 title claims abstract description 23
- 239000012535 impurity Substances 0.000 claims abstract description 35
- WEVYAHXRMPXWCK-UHFFFAOYSA-N acetonitrile Substances CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 22
- 239000012488 sample solution Substances 0.000 claims abstract description 18
- 238000001514 detection method Methods 0.000 claims abstract description 17
- HRQDCDQDOPSGBR-UHFFFAOYSA-M sodium;octane-1-sulfonate Chemical compound [Na+].CCCCCCCCS([O-])(=O)=O HRQDCDQDOPSGBR-UHFFFAOYSA-M 0.000 claims abstract description 16
- 239000000523 sample Substances 0.000 claims abstract description 9
- 239000000872 buffer Substances 0.000 claims abstract description 8
- 238000007865 diluting Methods 0.000 claims abstract description 6
- 239000007853 buffer solution Substances 0.000 claims abstract description 5
- 238000004007 reversed phase HPLC Methods 0.000 claims abstract description 3
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical compound C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 claims description 30
- 239000000243 solution Substances 0.000 claims description 27
- PCNDJXKNXGMECE-UHFFFAOYSA-N Phenazine Natural products C1=CC=CC2=NC3=CC=CC=C3N=C21 PCNDJXKNXGMECE-UHFFFAOYSA-N 0.000 claims description 15
- 238000012937 correction Methods 0.000 claims description 14
- 239000013558 reference substance Substances 0.000 claims description 11
- 239000002253 acid Substances 0.000 claims description 8
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 7
- 229910019142 PO4 Inorganic materials 0.000 claims description 6
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims description 6
- 239000010452 phosphate Substances 0.000 claims description 6
- 238000010812 external standard method Methods 0.000 claims description 5
- LFVGISIMTYGQHF-UHFFFAOYSA-N ammonium dihydrogen phosphate Chemical compound [NH4+].OP(O)([O-])=O LFVGISIMTYGQHF-UHFFFAOYSA-N 0.000 claims description 4
- 229910000387 ammonium dihydrogen phosphate Inorganic materials 0.000 claims description 4
- 235000019837 monoammonium phosphate Nutrition 0.000 claims description 4
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical group CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 3
- NQPDZGIKBAWPEJ-UHFFFAOYSA-N Valeric acid Natural products CCCCC(O)=O NQPDZGIKBAWPEJ-UHFFFAOYSA-N 0.000 claims description 2
- MNNHAPBLZZVQHP-UHFFFAOYSA-N diammonium hydrogen phosphate Chemical compound [NH4+].[NH4+].OP([O-])([O-])=O MNNHAPBLZZVQHP-UHFFFAOYSA-N 0.000 claims description 2
- 229910000388 diammonium phosphate Inorganic materials 0.000 claims description 2
- 235000019838 diammonium phosphate Nutrition 0.000 claims description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 2
- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 238000005259 measurement Methods 0.000 claims description 2
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 2
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 2
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims description 2
- 235000019799 monosodium phosphate Nutrition 0.000 claims description 2
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims description 2
- 238000009210 therapy by ultrasound Methods 0.000 claims description 2
- 229940005605 valeric acid Drugs 0.000 claims description 2
- XFXIJSRNFKHZFW-UHFFFAOYSA-N [Na].CCCCCCCC Chemical compound [Na].CCCCCCCC XFXIJSRNFKHZFW-UHFFFAOYSA-N 0.000 claims 2
- BDHFUVZGWQCTTF-UHFFFAOYSA-M sulfonate Chemical compound [O-]S(=O)=O BDHFUVZGWQCTTF-UHFFFAOYSA-M 0.000 claims 2
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 claims 1
- 239000007864 aqueous solution Substances 0.000 claims 1
- 229910052708 sodium Inorganic materials 0.000 claims 1
- 239000011734 sodium Substances 0.000 claims 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 abstract description 8
- 238000000926 separation method Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000006731 degradation reaction Methods 0.000 abstract description 3
- 239000012071 phase Substances 0.000 description 11
- 229960002749 aminolevulinic acid Drugs 0.000 description 9
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 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 5
- 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 5
- KSFOVUSSGSKXFI-GAQDCDSVSA-N CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O Chemical compound CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O KSFOVUSSGSKXFI-GAQDCDSVSA-N 0.000 description 5
- 229950003776 protoporphyrin Drugs 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000003085 diluting agent Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 238000009825 accumulation Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 150000003278 haem Chemical class 0.000 description 2
- 238000005286 illumination Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 239000006012 monoammonium phosphate Substances 0.000 description 2
- 239000003504 photosensitizing agent Substances 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 239000000377 silicon dioxide Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 239000003643 water by type Substances 0.000 description 2
- IXXIIAPIURLROR-UHFFFAOYSA-N 2-aminopentanoic acid;hydrochloride Chemical compound Cl.CCCC(N)C(O)=O IXXIIAPIURLROR-UHFFFAOYSA-N 0.000 description 1
- QGQXAMBOYWULFX-LZWSPWQCSA-N 2-morpholin-4-ylethyl (e)-6-(4,6-dihydroxy-7-methyl-3-oxo-1h-2-benzofuran-5-yl)-4-methylhex-4-enoate Chemical compound OC=1C=2C(=O)OCC=2C(C)=C(O)C=1C\C=C(/C)CCC(=O)OCCN1CCOCC1 QGQXAMBOYWULFX-LZWSPWQCSA-N 0.000 description 1
- ZCFFYALKHPIRKJ-UHFFFAOYSA-N 3-[18-(2-carboxylatoethyl)-8,13-bis(ethenyl)-3,7,12,17-tetramethyl-22,23-dihydroporphyrin-21,24-diium-2-yl]propanoate Chemical compound N1C(C=C2C(=C(C)C(=CC=3C(C)=C(CCC(O)=O)C(N=3)=C3)N2)C=C)=C(C)C(C=C)=C1C=C1C(C)=C(CCC(O)=O)C3=N1 ZCFFYALKHPIRKJ-UHFFFAOYSA-N 0.000 description 1
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 description 1
- ZLHFONARZHCSET-UHFFFAOYSA-N 5-aminolevulinic acid hydrochloride Chemical compound Cl.NCC(=O)CCC(O)=O ZLHFONARZHCSET-UHFFFAOYSA-N 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 206010034972 Photosensitivity reaction Diseases 0.000 description 1
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 1
- 239000012670 alkaline solution Substances 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000008238 biochemical pathway Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 231100000433 cytotoxic Toxicity 0.000 description 1
- 230000001472 cytotoxic effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005297 material degradation process Methods 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000012074 organic phase Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 238000012856 packing Methods 0.000 description 1
- 238000002428 photodynamic therapy Methods 0.000 description 1
- 230000036211 photosensitivity Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000027756 respiratory electron transport chain Effects 0.000 description 1
- -1 salt ion Chemical class 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
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- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for detecting substances related to aminolevulinic acid hydrochloride, which is a reversed-phase high-performance liquid chromatography method, and comprises the steps of preparing sodium octanesulfonate buffer solution with the pH value of 2.0+/-0.2, further preparing sodium octanesulfonate buffer solution-acetonitrile with acetonitrile accounting for 10% -20% (v/v) as a mobile phase, preparing a sample to be detected with the aminolevulinic acid hydrochloride as a main component into a sample solution with the concentration of 10mg/ml by using the mobile phase, taking 1ml of the sample solution, placing the sample solution into a 20ml measuring flask, diluting to a scale by using the mobile phase, shaking uniformly, and then detecting the sample solution. The detection method of the related substances of the amisole hydrochloride has good specificity, high detection sensitivity on each related substance, complete separation of impurity peaks and main peaks, more types of substances which can be detected, and 4 types of impurities which are generated in the degradation process of the amisole hydrochloride can be identified, wherein two types of the novel substances are novel substances which have not been identified before.
Description
Technical Field
The invention belongs to the technical field of medicine analysis, and particularly relates to a detection method of substances related to aminolevulinic acid hydrochloride.
Background
Aminopentanoic acid hydrochloride (Aminolevulinic Acid Hydrochloride), which has the chemical name 5-amino-4-oxopentanoic acid hydrochloride, formula: c (C) 5 H 9 NO 3 HCl, molecular weight: 167.59. the metabolism of aminolevulinic acid (ALA) is the first step in the biochemical pathway of heme synthesis in humans. ALA itself is not a photosensitizer, but a metabolic precursor of the photosensitizer protoporphyrin IX (PpIX). ALA synthesis is typically regulated by intracellular heme levels via ALA synthase feedback inhibition. Exogenous ALA circumvents this regulation, resulting in accumulation of PpIX. Skin is sensitized after application of amioxopentanoic acid hydrochloride by ALA conversion to PpIX and accumulation. When exposed to light of a certain wavelength and energy, the accumulated PpIX produces a photodynamic effect, a light and oxygen dependent cytotoxic effect: ppIX is in an excited state after absorbing light, which is then reacted with oxygenThe molecules undergo electron transfer to produce highly reactive singlet oxygen and free radicals. Photosensitivity of lesions after application of ALA with illumination of specific wavelengths and energies is the basis for ALA photodynamic therapy.
The aminolevulinic acid hydrochloride is unstable under alkaline solution and illumination conditions, and can be slowly degraded to generate impurities under high temperature conditions. In order to ensure the safety and effectiveness of the product, impurities need to be controlled, so that detection of related substances is an important index for product quality control.
To date, methods for detecting substances related to aminolevulinic acid hydrochloride have been only described in the united states pharmacopeia, and are described in the comparative examples below. The method has the following defects: (1) The amionovaleric acid hydrochloride has high polarity and can not be reserved in a reversed phase system, so that a main peak appears at the dead volume of a chromatographic column and can not be effectively separated from a solvent peak, and the chromatographic peak generates a shoulder peak phenomenon for a high-concentration sample solution. (2) The number of detected impurities is small, only two main impurities, namely 2, 5-dipropionate pyrazine (impurity A) and 3- (4' -carbonyl valeric acid) -2, 5-dipropionate pyrazine (impurity B), can be identified, and the other impurities can not be detected or positioned, even the unidentified impurities exist.
Therefore, it is necessary to optimize the detection method of the aminolevulinic acid hydrochloride related substance to obtain a method of detecting a larger variety of substances with higher sensitivity.
Disclosure of Invention
The invention aims to provide a method for detecting substances related to aminolevulinic acid hydrochloride with higher sensitivity.
The aim of the invention is achieved by the following technical scheme:
the invention provides a detection method of a related substance of aminolevulinic acid hydrochloride, which is a reversed-phase high-performance liquid chromatography method and comprises the following steps:
(1) Preparing a solution:
a) Preparing sodium octane sulfonate buffer solution with pH value of 2.0 plus or minus 0.2, and further preparing sodium octane sulfonate buffer solution-acetonitrile with acetonitrile accounting for 10-20% (v/v) as mobile phase;
b) Preparing a sample to be detected with the aminolevulinic acid hydrochloride as a main component into a sample solution with the concentration of 10mg/ml by using the mobile phase;
c) Putting 1ml of the sample solution into a 20ml measuring flask, diluting to a scale with the mobile phase, shaking uniformly, putting 5ml of the sample solution into a 50ml measuring flask, diluting to the scale with the mobile phase, shaking uniformly, and taking as a control solution;
d) Taking 2, 5-dipropionate pyrazine (hereinafter referred to as impurity A) as a reference substance, and adding the mobile phase after ultrasonic treatment for 30 seconds by using 50% acetonitrile water solution to dissolve the reference substance to prepare a reference substance solution with the concentration of 20 mug/ml;
(2) Chromatographic conditions: the chromatographic column filler is octadecylsilane chemically bonded silica gel; the detection wavelength is ultraviolet wavelength; the column temperature is 25-30 ℃; the flow rate is 0.5 ml/min-1.0 ml/min;
(3) And (3) measuring and calculating:
a) Respectively taking 20 mu l of each of the sample solution, the control solution and the control solution, and injecting the sample solution into a liquid chromatograph for carrying out the same method measurement to obtain a sample solution chromatogram, a control solution chromatogram and a control solution chromatogram;
b) Respectively calculating the content of each related substance in the chromatogram of the solution of the sample to be tested by taking the impurity A as a reference substance and an external standard method of adding correction factors;
c) The correction factors for each relevant substance are as follows:
the correction factor of impurity a is 1;
3- (4' -carbonylvaleric acid) -2, 5-dipropionic acid pyrazine (hereinafter referred to as impurity B) correction factor of 1.8; 3-methyl-2, 5-dipropionate pyrazine (hereinafter referred to as impurity C) correction factor of 1.2;
the correction factor of 3-ethyl-2, 5-dipropionate pyrazine (hereinafter referred to as impurity D) was 1.3;
calculating correction factors of unknown related substances i according to Ai/At0.5%, wherein Ai is the peak area of the unknown related substances i; at is the main peak area in the control solution chromatogram.
Further, in the sodium octanesulfonate buffer solution, the concentration of sodium octanesulfonate is 15 mM-30 mM; the concentration of phosphate is 15 mM-70 mM.
Preferably, in the sodium octanesulfonate buffer, the concentration of sodium octanesulfonate is 20mM, and the concentration of phosphate is 40mM.
Further, the phosphate is at least one selected from potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate, and diammonium hydrogen phosphate.
Further, acetonitrile in the mobile phase accounts for 15%.
Further, the detection wavelength is 204nm to 206nm.
Further, the flow rate was 1.0ml/min.
The invention has the outstanding effects that:
the detection method of the related substances of the amisole hydrochloride has good specificity, high detection sensitivity on each related substance, complete separation of impurity peaks and main peaks can be realized, the detected substances are more, 4 kinds of impurities generated in the degradation process of the amisole hydrochloride can be identified, and the impurity C and the impurity D are new substances which have not been identified before.
In conclusion, the impurity detection method is adopted in the production of the aminolevulinic acid hydrochloride, so that the product quality control level can be greatly improved, and the method has great significance for industrial progress.
Drawings
FIG. 1 is a chromatogram of example 1 of the present invention;
FIG. 2 is a chromatogram of example 2 of the present invention;
FIG. 3 is a chromatogram of example 3 of the present invention;
FIG. 4 is a chromatogram of example 4 of the present invention;
FIG. 5 is a chromatogram of example 5 of the present invention;
FIG. 6 is a chromatogram of example 6 of the present invention;
FIG. 7 is a chromatogram of example 7 of the present invention;
FIG. 8 is a chromatogram of example 8 of the present invention;
FIG. 9 is a chromatogram of example 9 of the present invention;
FIG. 10 is a chromatogram of example 10 of the present invention;
FIG. 11 is a chromatogram of example 11 of the present invention;
FIG. 12 is a chromatogram of a comparative example of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. It is to be understood that these examples are illustrative of the present invention and are not intended to limit the scope of the present invention. Further, it is understood that various changes and modifications may be made by those skilled in the art after reading the teachings of the present invention, and such equivalents are intended to fall within the scope of the claims appended hereto.
Examples 1 to 10
(1) Experimental apparatus and reagent
Instrument: agilent HPLC 1260 Method Scouting system
(2) Chromatographic conditions:
chromatographic column: octadecylsilane chemically bonded silica is the packing of the column, agilent Zorbax C18.6X105 mm 5 μm. The chromatographic conditions of examples 1 to 10 are shown in Table 1 below.
Table 1: chromatographic conditions of examples 1 to 10
Note that: mM is an abbreviation for concentration unit mmol/L, 1mM = 1X 103mol/L.
(3) Sample preparation: the sodium octanesulfonate buffer solution and the mobile phase are prepared according to the proportion of the table 2, then a proper amount of the aminolevulinic acid hydrochloride bulk drug is weighed, and the mobile phase is used for preparing the solution with the concentration of 10 mg/ml.
(4) Experimental results:
the chromatograms obtained in examples 1 to 10 were shown in FIGS. 1 to 10, respectively, and the main component was well separated from impurity A, impurity B, impurity C and impurity D, which were new substances that were not identified before.
As shown in the following table 2, the detection method disclosed by the invention identifies 4 impurities generated in the degradation process of the aminolevulinic acid hydrochloride as follows: impurity A:2, 5-dipropionate pyrazine;
impurity B:3- (4' -carbonylvaleric acid) -2, 5-dipropionic acid pyrazine;
impurity C: 3-methyl-2, 5-dipropionate pyrazine;
impurity D: 3-ethyl-2, 5-dipropionate pyrazine.
TABLE 24 impurities identifiable by the detection method disclosed in the present invention
Example 11
(1) Experimental apparatus and reagent
Instrument: same as in examples 1 to 10
(2) Chromatographic conditions:
chromatographic column: same as in examples 1 to 10
Flow rate: 1.0ml/min
Detection wavelength: 205nm
Column temperature: 30 DEG C
Sample injection amount: 20 mu L
Mobile phase: sodium octanesulfonate buffer-acetonitrile=86:14 (v/v) (the sodium octanesulfonate buffer is prepared by taking 4.75g (20 mM) of sodium octanesulfonate, 7.59g (66 mM) of monoammonium phosphate, adding 980ml of water to dissolve, adjusting the pH value to 1.90 with phosphoric acid, and diluting to 1000ml with water.
(3) Sample preparation: the proper amount of aminolevulinic acid hydrochloride preparation containing auxiliary materials and auxiliary material degradation impurities (parahydroxybenzoic acid) is weighed, and the aminolevulinic acid hydrochloride preparation is prepared into a solution with the concentration of 10mg/ml by using a mobile phase.
(4) Experimental results:
the chromatogram obtained in example 11 is shown in FIG. 11.
Further, DOE robustness test was performed on example 11 by statistical software Minitab 17 (ID: 30034556135249531400), and DOE parameters were designed as follows:
factor (2):
salt ion pair concentration = 86mM to 90mM (i.e. 21.5mM to 22.5mM sodium octane sulfonate + 64.5mM to 67.5mM monoammonium phosphate);
buffer ph=1.85-1.95;
organic phase ratio = 13.5% -14.5%;
flow rate = 0.95 ml/min-1.05 ml/min;
column temperature = 28 ℃ -32 ℃;
and (3) responding: r1 (degree of separation of main peak from impurity D); r2 (degree of separation of impurity E from main peak); r3 (degree of separation of impurity F from impurity E); a main peak tailing factor; number of theoretical plates of main peak.
DOE model: five factors two levels (3 center points, i.e. center points repeated three times)
DOE robustness test results: referring to Table 3, the results show that R1>1.0, R2>3.0, R3>2.0, and Tailing <3.0, platecount >3000 in the DOE parameter range, and the method has good robustness for detecting related substances of the amionovaleric acid hydrochloride preparation containing auxiliary materials in the above parameter range.
Table 3: DOE robustness test results
Comparative example: method for detecting American pharmacopoeia records
(1) Experimental apparatus and reagent
Instrument: waters UPLC
(2) Chromatographic conditions:
chromatographic column: octadecylsilane chemically bonded silica is used as filler of chromatographic column, waters BEH Shield RP, 2.1X100 mm, 1.7 μm
Flow rate: 0.4ml/min
Detection wavelength: 210nm of
Sample injection amount: 5 mu L
Solution a:2M sulfuric acid adjusts the pH value of the water to 2.2
A diluent: methanol and solution A (1:3) (v/v)
Elution procedure: see table 4.
Table 4: elution procedure of comparative example
| Time min | Solution A% | Methanol% |
| 0 | 95 | 5 |
| 2 | 95 | 5 |
| 6 | 60 | 40 |
| 8 | 60 | 40 |
| 9 | 95 | 5 |
| 23 | 95 | 5 |
(3) Sample solution preparation: an appropriate amount of aminolevulinic acid hydrochloride was weighed and formulated with a diluent into a solution having a concentration of 40mg/ml and 40 μg/ml.
(4) Preparing a reference substance solution: the proper amount of the aminolevulinic acid hydrochloride, the impurity A and the impurity B are respectively weighed and respectively prepared into 0.04mg/ml solution by using a diluent.
(5) Experimental results:
the impurities A and B are calculated according to an external standard method by respective reference substances; calculating other single unknown impurities which are before the impurity A by taking the aminolevulinic acid hydrochloride as a reference substance according to an external standard method; the other single unknown impurity which shows a peak after the related impurity A is calculated by an external standard method by taking the related impurity A as a reference substance.
As shown in FIG. 12, the obtained chromatogram shows that the peak of the aminolevulinic acid hydrochloride is shoulder peak at the concentration of 40mg/ml, and the peak is generated at the dead volume position and cannot be effectively separated from the solvent peak.
While particular embodiments of the present invention have been described above, it will be appreciated by those skilled in the art that these are merely illustrative, and that many changes and modifications may be made to these embodiments without departing from the principles and spirit of the invention. Accordingly, the scope of the invention is defined by the appended claims.
Claims (7)
1. A method for detecting substances related to aminolevulinic acid hydrochloride, which is a reversed-phase high-performance liquid chromatography, and is characterized by comprising the following steps:
preparing a solution:
preparing an octane sodium sulfonate buffer solution with the pH value of 2.0+/-0.2, and further preparing an octane sodium sulfonate buffer solution-acetonitrile with the acetonitrile content of 10% -20% (v/v) as a mobile phase;
preparing a sample to be detected with the aminolevulinic acid hydrochloride as a main component into a sample solution with the concentration of 10mg/ml by using the mobile phase;
putting 1ml of the sample solution into a 20ml measuring flask, diluting to a scale with the mobile phase, shaking uniformly, putting 5ml of the sample solution into a 50ml measuring flask, diluting to the scale with the mobile phase, shaking uniformly, and taking as a control solution;
taking 2, 5-dipropionate pyrazine as a reference substance, and adding the mobile phase after ultrasonic treatment for 30 seconds by using a 50% acetonitrile aqueous solution to dissolve the pyrazine to prepare a reference substance solution with the concentration of 20 mu g/ml;
chromatographic conditions: the chromatographic column filler is octadecylsilane chemically bonded silica gel; the detection wavelength is ultraviolet wavelength; the column temperature is 25-30 ℃; the flow rate is 0.5 ml/min-1.0 ml/min;
and (3) measuring and calculating:
respectively taking 20 mu l of each of the sample solution, the control solution and the control solution, and injecting the sample solution into a liquid chromatograph for carrying out the same method measurement to obtain a sample solution chromatogram, a control solution chromatogram and a control solution chromatogram;
respectively calculating the content of each related substance in the chromatogram of the solution of the sample to be tested by taking the impurity A as a reference substance and an external standard method of adding correction factors;
the correction factors for each relevant substance are as follows:
the correction factor of impurity a is 1;
3- (4' -carbonyl valeric acid) -2, 5-dipropionic acid pyrazine correction factor 1.8;
the correction factor of 3-methyl-2, 5-dipropionate pyrazine is 1.2;
the correction factor of 3-ethyl-2, 5-dipropionate pyrazine is 1.3;
calculating correction factors of unknown related substances i according to Ai/At0.5%, wherein Ai is the peak area of the unknown related substances i; at is the main peak area in the control solution chromatogram.
2. The method according to claim 1, wherein the concentration of sodium alkanesulfonate in the sodium octanesulfonate buffer is 15 mM-30 mM; the concentration of phosphate is 15 mM-70 mM.
3. The method according to claim 2, wherein the concentration of sodium octanesulfonate in the sodium octanesulfonate buffer is 20mM and the concentration of phosphate is 40mM.
4. The method according to claim 2 or 3, wherein the phosphate is at least one selected from potassium dihydrogen phosphate, sodium dihydrogen phosphate, ammonium dihydrogen phosphate, dipotassium hydrogen phosphate, disodium hydrogen phosphate and diammonium hydrogen phosphate.
5. The method of claim 1, wherein the acetonitrile in the mobile phase is 15%.
6. The method according to claim 1, wherein the detection wavelength is 204nm to 206nm.
7. The method of claim 1, wherein the flow rate is 1.0ml/min.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106226421A (en) * | 2016-07-14 | 2016-12-14 | 合肥华方医药科技有限公司 | A kind of Sorafenib Tosylate has the detection method of related substance |
| CN107976508A (en) * | 2017-11-27 | 2018-05-01 | 上海百灵医药科技有限公司 | A kind of method for analyzing hydrochloric acid ammonia ketone valeric acid content and purity |
| CN109490460A (en) * | 2018-12-27 | 2019-03-19 | 广东盛泰华生物制药有限公司 | A kind of detection method of the L-Arginine in relation to substance |
| CN111592484A (en) * | 2020-06-06 | 2020-08-28 | 上海奈及生物医药科技有限公司 | Preparation method of 5-aminolevulinic acid hydrochloride intermediate |
-
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106226421A (en) * | 2016-07-14 | 2016-12-14 | 合肥华方医药科技有限公司 | A kind of Sorafenib Tosylate has the detection method of related substance |
| CN107976508A (en) * | 2017-11-27 | 2018-05-01 | 上海百灵医药科技有限公司 | A kind of method for analyzing hydrochloric acid ammonia ketone valeric acid content and purity |
| CN109490460A (en) * | 2018-12-27 | 2019-03-19 | 广东盛泰华生物制药有限公司 | A kind of detection method of the L-Arginine in relation to substance |
| CN111592484A (en) * | 2020-06-06 | 2020-08-28 | 上海奈及生物医药科技有限公司 | Preparation method of 5-aminolevulinic acid hydrochloride intermediate |
Non-Patent Citations (1)
| Title |
|---|
| 毫火针与毫针对5-盐酸氨酮戊酸体外经皮渗透的影响研究;陈茜;高燕妮;杨富金;刁庆春;陶春蓉;黄筠;;四川中医(07);第52-55页 * |
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