CN107367562A - The analyzing detecting method of sulfuric acid Polymyxin B sulfate and application - Google Patents
The analyzing detecting method of sulfuric acid Polymyxin B sulfate and application Download PDFInfo
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- CN107367562A CN107367562A CN201710656465.6A CN201710656465A CN107367562A CN 107367562 A CN107367562 A CN 107367562A CN 201710656465 A CN201710656465 A CN 201710656465A CN 107367562 A CN107367562 A CN 107367562A
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- polymyxin
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- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 title claims abstract description 197
- 108010093965 Polymyxin B Proteins 0.000 title claims abstract description 117
- SBKRTALNRRAOJP-BWSIXKJUSA-N N-[(2S)-4-amino-1-[[(2S,3R)-1-[[(2S)-4-amino-1-oxo-1-[[(3S,6S,9S,12S,15R,18R,21S)-6,9,18-tris(2-aminoethyl)-15-benzyl-3-[(1R)-1-hydroxyethyl]-12-(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-1-oxobutan-2-yl]-6-methylheptanamide (6S)-N-[(2S)-4-amino-1-[[(2S,3R)-1-[[(2S)-4-amino-1-oxo-1-[[(3S,6S,9S,12S,15R,18R,21S)-6,9,18-tris(2-aminoethyl)-15-benzyl-3-[(1R)-1-hydroxyethyl]-12-(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-hydroxy-1-oxobutan-2-yl]amino]-1-oxobutan-2-yl]-6-methyloctanamide sulfuric acid Polymers OS(O)(=O)=O.CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@@H](NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](Cc2ccccc2)NC(=O)[C@@H](CCN)NC1=O)[C@@H](C)O.CC[C@H](C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@@H](NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](Cc2ccccc2)NC(=O)[C@@H](CCN)NC1=O)[C@@H](C)O SBKRTALNRRAOJP-BWSIXKJUSA-N 0.000 title claims abstract description 89
- 229960003548 polymyxin b sulfate Drugs 0.000 title claims abstract description 89
- 238000000034 method Methods 0.000 title claims abstract description 44
- 238000004128 high performance liquid chromatography Methods 0.000 claims abstract description 29
- 238000002360 preparation method Methods 0.000 claims abstract description 25
- 238000001514 detection method Methods 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 19
- 229940079593 drug Drugs 0.000 claims abstract description 17
- 238000000105 evaporative light scattering detection Methods 0.000 claims abstract description 17
- 238000004458 analytical method Methods 0.000 claims abstract description 15
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 claims description 38
- 239000000243 solution Substances 0.000 claims description 32
- 239000013558 reference substance Substances 0.000 claims description 31
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 30
- 239000012071 phase Substances 0.000 claims description 20
- 239000007864 aqueous solution Substances 0.000 claims description 17
- 239000007788 liquid Substances 0.000 claims description 12
- 239000003495 polar organic solvent Substances 0.000 claims description 8
- 238000002347 injection Methods 0.000 claims description 7
- 239000007924 injection Substances 0.000 claims description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical group OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 239000003643 water by type Substances 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 239000006199 nebulizer Substances 0.000 claims description 5
- 238000000889 atomisation Methods 0.000 claims description 3
- 239000000945 filler Substances 0.000 claims description 2
- 239000007791 liquid phase Substances 0.000 claims description 2
- -1 octadecyl Silica gel Chemical compound 0.000 claims description 2
- 239000002245 particle Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000005070 sampling Methods 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims description 2
- 238000004611 spectroscopical analysis Methods 0.000 claims 1
- 230000035945 sensitivity Effects 0.000 abstract description 6
- 230000000694 effects Effects 0.000 abstract description 4
- 239000012535 impurity Substances 0.000 abstract description 4
- 229920000024 polymyxin B Polymers 0.000 description 28
- 229960005266 polymyxin b Drugs 0.000 description 28
- 238000002474 experimental method Methods 0.000 description 16
- 230000000052 comparative effect Effects 0.000 description 11
- 239000000203 mixture Substances 0.000 description 10
- 239000008346 aqueous phase Substances 0.000 description 8
- 239000002904 solvent Substances 0.000 description 8
- 238000012360 testing method Methods 0.000 description 6
- PBCJIPOGFJYBJE-UHFFFAOYSA-N acetonitrile;hydrate Chemical compound O.CC#N PBCJIPOGFJYBJE-UHFFFAOYSA-N 0.000 description 5
- KNIWPHSUTGNZST-UHFFFAOYSA-N polymyxin E2 Natural products CC(C)CCCCC(=O)NC(CCN)C(=O)NC(C(C)O)C(=O)NC(CCN)C(=O)NC1CCNC(=O)C(C(C)O)NC(=O)C(CCN)NC(=O)C(CCN)NC(=O)C(CC(C)C)NC(=O)C(CC(C)C)NC(=O)C(CCN)NC1=O KNIWPHSUTGNZST-UHFFFAOYSA-N 0.000 description 5
- 108010078777 Colistin Proteins 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 229960003346 colistin Drugs 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- JORAUNFTUVJTNG-BSTBCYLQSA-N n-[(2s)-4-amino-1-[[(2s,3r)-1-[[(2s)-4-amino-1-oxo-1-[[(3s,6s,9s,12s,15r,18s,21s)-6,9,18-tris(2-aminoethyl)-3-[(1r)-1-hydroxyethyl]-12,15-bis(2-methylpropyl)-2,5,8,11,14,17,20-heptaoxo-1,4,7,10,13,16,19-heptazacyclotricos-21-yl]amino]butan-2-yl]amino]-3-h Chemical compound CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O.CCC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@H]([C@@H](C)O)CN[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O JORAUNFTUVJTNG-BSTBCYLQSA-N 0.000 description 4
- XDJYMJULXQKGMM-UHFFFAOYSA-N polymyxin E1 Natural products CCC(C)CCCCC(=O)NC(CCN)C(=O)NC(C(C)O)C(=O)NC(CCN)C(=O)NC1CCNC(=O)C(C(C)O)NC(=O)C(CCN)NC(=O)C(CCN)NC(=O)C(CC(C)C)NC(=O)C(CC(C)C)NC(=O)C(CCN)NC1=O XDJYMJULXQKGMM-UHFFFAOYSA-N 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 238000004587 chromatography analysis Methods 0.000 description 3
- 238000003908 quality control method Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 101000777206 Homo sapiens Ubiquitin carboxyl-terminal hydrolase 40 Proteins 0.000 description 2
- 108010040201 Polymyxins Proteins 0.000 description 2
- 102100031284 Ubiquitin carboxyl-terminal hydrolase 40 Human genes 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000010354 integration Effects 0.000 description 2
- 238000010829 isocratic elution Methods 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 238000004445 quantitative analysis Methods 0.000 description 2
- LCPVQAHEFVXVKT-UHFFFAOYSA-N 2-(2,4-difluorophenoxy)pyridin-3-amine Chemical compound NC1=CC=CN=C1OC1=CC=C(F)C=C1F LCPVQAHEFVXVKT-UHFFFAOYSA-N 0.000 description 1
- USFZMSVCRYTOJT-UHFFFAOYSA-N Ammonium acetate Chemical compound N.CC(O)=O USFZMSVCRYTOJT-UHFFFAOYSA-N 0.000 description 1
- 239000005695 Ammonium acetate Substances 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- SJGCVQNEYOPPLH-UHFFFAOYSA-L C(C)#N.O.P(O)(O)(O)=O.S(=O)(=O)([O-])[O-].[Na+].[Na+] Chemical compound C(C)#N.O.P(O)(O)(O)=O.S(=O)(=O)([O-])[O-].[Na+].[Na+] SJGCVQNEYOPPLH-UHFFFAOYSA-L 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000589517 Pseudomonas aeruginosa Species 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- 206010072170 Skin wound Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 235000019257 ammonium acetate Nutrition 0.000 description 1
- 229940043376 ammonium acetate Drugs 0.000 description 1
- VZTDIZULWFCMLS-UHFFFAOYSA-N ammonium formate Chemical compound [NH4+].[O-]C=O VZTDIZULWFCMLS-UHFFFAOYSA-N 0.000 description 1
- 230000002924 anti-infective effect Effects 0.000 description 1
- 239000011260 aqueous acid Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 241000385736 bacterium B Species 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- KNIWPHSUTGNZST-SSWRVQTPSA-N colistin B Chemical compound CC(C)CCCCC(=O)N[C@@H](CCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCN)C(=O)N[C@H]1CCNC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCN)NC(=O)[C@H](CCN)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](CC(C)C)NC(=O)[C@H](CCN)NC1=O KNIWPHSUTGNZST-SSWRVQTPSA-N 0.000 description 1
- 238000000205 computational method Methods 0.000 description 1
- 238000010219 correlation analysis Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000001704 evaporation Methods 0.000 description 1
- 230000008020 evaporation Effects 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 150000004665 fatty acids Chemical group 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 150000002825 nitriles Chemical class 0.000 description 1
- 206010034674 peritonitis Diseases 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- CHQMHPLRPQMAMX-UHFFFAOYSA-L sodium persulfate Substances [Na+].[Na+].[O-]S(=O)(=O)OOS([O-])(=O)=O CHQMHPLRPQMAMX-UHFFFAOYSA-L 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000002485 urinary effect Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a kind of analyzing detecting method of sulfuric acid Polymyxin B sulfate.The detection method comprises the steps of：Evaporative light-scattering device is combined using high performance liquid chromatography, analysis detection is carried out to sulfuric acid Polymyxin B sulfate, you can.It the method achieve and be kept completely separate four components in sulfuric acid Polymyxin B sulfate, specificity is strong, it is simple to operate, and do not disturbed by other organic impurities, the HPLC methods of effect and the bulk drugs of EP 9.0 are close, high sensitivity, simple and quick, can more accurately be used to determine each component content in sulfuric acid Polymyxin B sulfate preparation.
Description
Technical field
The present invention relates to sulfuric acid Polymyxin B sulfate analyzing detecting method and application.
Background technology
Polymyxin B sulfate (polymyxin B) forms caused by more glutinous bacillus, by several amino acids and aliphatic acid
A kind of basic polypeptide class antibiotic.Current production mainly uses microbe fermentation method biosynthesis, then extracted is refining to obtain
Arrive.Its sulfate of Clinical practice, used mainly as anti-infection drug, treat and caused by Gram-negative bacterias such as Pseudomonas aeruginosas
Skin wound, urinary system and eye, ear, the site infection such as tracheae, it can also be used to septicemia, peritonitis etc..Sulfuric acid is more glutinous
Rhzomorph B is the similar multicomponent mixture of structure, including Polymyxin B sulfate1、B2、B3、B1-I.Wherein, B1And B2It is active ingredient,
B3And B1- I is major impurity.Chemical structural formula is as follows：
Specifically, Polymyxin B sulfate1、B2、B3And B1Difference in-I structures is as follows：
At present, it is more glutinous to have recorded sulfuric acid by European Pharmacopoeia EP 9.0, American Pharmacopeia USP 40 and Chinese Pharmacopoeia ChP 2015
Rhzomorph B or its preparation correlation analysis detection method.
Wherein, sulfuric acid Polymyxin B sulfate assay uses the anti-phase of sodium sulphate-phosphoric acid-water-acetonitrile system in EP 9.0
High performance liquid chromatography isocratic elution, detect at ultraviolet wavelength 215nm, external standard method calculates each component content, single needle analysis time
About 50min.Detection method without related preparations；
Though USP 40 and ChP 2015 calculates containing for sulfuric acid Polymyxin B sulfate and its preparation using microbial antibiotic potency method
Amount, but the high performance liquid chromatography consistent with EP 9.0 is also used under polymyxin component check item, and be required to dry
Dry product meter, B1, B2, B3, B1-I summation >=80.0%, B3≤6.0%, B1-I≤15.0%；
Orwa etc. is using acetonitrile-aqueous sodium persulfate solution (0.7%, m/v)-phosphate aqueous solution (5.8%, v/v)-water
(22.25：50：5：22.75) mobile phase based on, the sulfuric acid Polymyxin B sulfate in the case where 215nm has investigated different pH1、B2、B3Separation
Situation (Journal of Chromatography A, 870 (2000) 237-243).
Because each group separation structure in sulfuric acid Polymyxin B sulfate is similar, only there is area in fatty acid chain and Z amino acids residues
Not, and each one pack system content difference it is big.In addition sulfuric acid Polymyxin B sulfate absorbs weaker under 215nm, each during using the above method
The signal intensity of component is low, so the accuracy of chromatographic integration during quantitative analysis can be influenceed to a certain degree；Also, isocratic elution side
Method also has the shortcomings that to cause that the post effect for separating each component is low, analysis time is longer.Therefore, exploitation it is a kind of it is quick, sensitive, accurate,
The method that quantitative analysis can be carried out to each component simultaneously, the quality control to medicine are significant.
The content of the invention
Technical problem solved by the invention is to survey using antibiotic microorganism potency method in the prior art to overcome
Determine that specificity is poor during sulfuric acid Polymyxin B sulfate, precision is low or more to sulfuric acid using high performance liquid chromatography combination UV-detector
When colistin B carries out analysis detection, exist mobile phase prepare complicated, sensitivity is low, poor repeatability, long analysis time the problems such as,
So as to provide analyzing detecting method and its application that a kind of sulfuric acid sticks plain bacterium B more, the analysis method is relative to the micro- life of antibiotic
Thing potency method specificity is simple to operate by force, can be kept completely separate four components, and not by other organic impurities disturb, effect with
The HPLC methods of the bulk drugs of EP 9.0 are close, high sensitivity, simple and quick, can more accurately be used to determine sulfuric acid Polymyxin B sulfate
Each component content in preparation.
The present invention is that solve above-mentioned technical problem by the following technical programs.
The invention provides a kind of analyzing detecting method of sulfuric acid Polymyxin B sulfate, it is comprised the steps of：Using efficient liquid
Phase chromatography combines evaporative light-scattering device, and analysis detection is carried out to sulfuric acid Polymyxin B sulfate, you can；
Chromatographic column in described high performance liquid chromatography is inverse analysis post, and chromatographic column fixed phase is bonded for octadecyl
Silica gel；Such as Kromasil 100-5C18 posts；
Mobile phase in described high performance liquid chromatography is polar organic solvent and trifluoroacetic acid aqueous solution, the trifluoro
The volumn concentration of trifluoroacetic acid is 0.01%~0.3% in acetic acid aqueous solution；Trifluoro second in the trifluoroacetic acid aqueous solution
The computational methods of the volumn concentration of acid account for the ratio of trifluoroacetic acid aqueous solution cumulative volume for the volume of trifluoroacetic acid.
In the present invention, the sulfuric acid Polymyxin B sulfate is Polymyxin B sulfate1、B2、B3And B1- I mixtures.
In the present invention, described high performance liquid chromatography can use gradient elution, the trifluoro second during gradient elution
The volume ratio of aqueous acid and the polar organic solvent can be 85:15~60:40.
In the present invention, in the gradient elution, the mobile phase ratio changes with time scope can be as shown in table 1：
Table 1
Again can be as shown in table 2 or table 3：
Table 2
| Time (min) | Polar organic solvent (%, v/v) |
| 0 | 24 |
| 5.00 | 24 |
| 5.01~8.00 | 24~29 |
| 8.01~15.00 | 29~32 |
| 15.01~20.00 | 32~40 |
Table 3
| Time (min) | Polar organic solvent (%, v/v) |
| 0 | 24 |
| 5.00 | 24 |
| 5.01~8.01 | 24~27 |
| 8.01~15.00 | 27~30 |
| 15.01~20.00 | 30~40 |
In the present invention, the volumn concentration of the trifluoroacetic acid in the trifluoroacetic acid aqueous solution can be 0.05%~
0.1%.
In the present invention, the polar organic solvent can use the polarity commonly used in this area in high performance liquid chromatography organic molten
One or more in agent, such as methanol, ethanol and acetonitrile.
In the present invention, the high performance liquid chromatograph in the high performance liquid chromatography can use the conventional high-performance liquid of this area
Chromatography, such as the model Alliance e2695 high performance liquid chromatographs that producer is Waters.
In the present invention, the specification of the chromatographic column can be this area conventional analysis post specification, and the color of following specification may be selected
Compose post：The filler particles degree of the chromatographic column can be 2.7 μm~5 μm；Such as 3 μm~5 μm；The length of the chromatographic column can be
100mm~250mm, such as 150mm；The sampling volume of the chromatographic column can be 1~50 μ L, such as 20 μ L.
In the present invention, the column temperature in the high performance liquid chromatography can be the ordinary temperature of this area generic operation, such as
20~40 DEG C, then such as 35 DEG C.
In the present invention, the flow velocity of described mobile phase refers to the routine operation in field, such as 0.5~1.5mL/min, then
Such as 0.8~1.0mL/min.
In the present invention, the evaporative light-scattering device can be using this area routine and evaporation associated with high performance liquid chromatograph
Diffuser, such as the evaporative light-scattering device for the model 2424 that producer is Waters, the parameter of the evaporative light-scattering device are set
The mode of putting can be conventional arrangement mode, such as：
The drift tube temperature of the evaporative light-scattering device can be 40~80 DEG C；Such as 60~70 DEG C；
The nebulizer gas pressure of the evaporative light-scattering device can be 20~45psi；Such as 25~35psi；
The atomization mouth temperature of the evaporative light-scattering device can be 12 DEG C~35 DEG C；Such as 12~30 DEG C.
In the present invention, described sulfuric acid sticks plain B more and also can be replaced the bulk drug that sulfuric acid sticks plain B more, or sulfuric acid sticks more
Plain B preparation, or sulfuric acid stick the mixture of two or more component of B1, B2, B3, B1-I in plain B more.
Present invention also offers the analyzing detecting method in sulfuric acid Polymyxin B sulfate bulk drug or formulation content detection
Using it is comprised the steps of：
Step (1), certain density reference substance solution and sulfuric acid Polymyxin B sulfate bulk drug or preparation product to be tested is respectively configured
Solution, the reference substance solution and the sulfuric acid Polymyxin B sulfate preparation product to be tested solution are entered using above-mentioned analyzing detecting method
Row analysis detection；Measure the peak area of each component in the reference substance solution and the sulfuric acid Polymyxin B sulfate preparation；
Step (2), the peak area of each component and corresponding in sulfuric acid Polymyxin B sulfate according to the reference substance solution
Concentration of component, obtain regression equation；
The content of each component of step (3), the sulfuric acid Polymyxin B sulfate bulk drug or preparation is by measuring in step (1)
The regression equation obtained in the substitution step (2) of the peak area of each component, you can.
The preparation can be injection sterile powder.
In the step (1), the reference substance can be sulfuric acid Polymyxin B sulfate USP reference substances, for example, lot number is
N1M425, often contain B in reference substance described in 100mg1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg；
In the step (1), the preparation of the reference substance solution and the sulfuric acid Polymyxin B sulfate preparation product to be tested solution can
Using the Conventional solvents that can be dissolved, such as the mixed solvent of acetonitrile and water, such as the volume ratio of acetonitrile and water is 8:2 it is mixed
Bonding solvent；
The concentration of the reference substance solution is referred to when this area makes the double-log regression equation of concentration and peak area
Concentration, such as 0.3mg/mL~1.5mg/mL；The concentration of the sulfuric acid Polymyxin B sulfate preparation product to be tested solution can be according to reference substance
The scope of concentration is selected, such as 0.5mg/mL.
In the step (2), the acquisition of the regression equation refers to the conventional Calculation Method of this area, for example, with institute
Stating the peak area of each component in sulfuric acid Polymyxin B sulfate described in reference substance solution, to take 10 logarithms for being bottom be abscissa, with corresponding
It is ordinate that the mass concentration of component, which takes 10 logarithms for being bottom, is fitted conic section, calculates to obtain the regression equation.
In the step (3), the content of each component of the sulfuric acid Polymyxin B sulfate preparation can be by measuring in step (1)
The peak area of each component takes 10 to substitute into step (2) obtained regression equation for the logarithm at bottom.
Application of the analyzing detecting method in the detection of sulfuric acid Polymyxin B sulfate formulation content, can be comprised the steps of：
(1) weigh sulfuric acid Polymyxin B sulfate reference substance (for mixture known to each component content, by taking USP reference substances as an example,
69.7mg containing B1, B2 15.9mg, B3 3.2mg, B1-I 6.1mg in per 100mg) in right amount, it is placed in volumetric flask, with water-second
Nitrile (80:20) make solvent to dissolve and be diluted to scale, the total concentration for obtaining sulfuric acid Polymyxin B sulfate is 5.0mg/mL solution, is made
For reference substance solution A；Take from solution A and be respectively placed in right amount in volumetric flask, be configured to total concentration be respectively 0.3mg/mL~
1.5mg/mL reference substance solution；
Take sulfuric acid Polymyxin B sulfate bulk drug or preparation test sample appropriate, with water-acetonitrile (80:20, volume ratio) match somebody with somebody for solvent
The need testing solution that concentration is 0.5mg/mL is made；
(2) reference substance solution and each 20 μ L sample introductions of need testing solution are taken, using above-mentioned analyzing detecting method, according to institute
The peak area of sulfuric acid Polymyxin B sulfate each component and corresponding solution component mass concentration in reference substance solution are stated, is each taken the logarithm
After be fitted conic section；
(3) content of each component substitutes into standard curve calculating by the logarithm of the peak area measured in the need testing solution
；Sulfuric acid Polymyxin B sulfate each component content sum is total content.
Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can be combined, and it is each preferably to produce the present invention
Example.
Agents useful for same and instrument of the present invention are commercially available.
The positive effect of the present invention is：
(1) analyzing detecting method has the characteristics that strong simple to operate, specificity, high sensitivity and simple and quick.
(2) analyzing detecting method provides for Polymyxin B sulfate each component log concentration-peak area logarithm in pharmacopoeia of each country
Quality control clearance in have it is good linear；
(3) analyzing detecting method identification of each component or quantitative detection suitable for sulfuric acid Polymyxin B sulfate, it can also be used to
The content detection of sulfuric acid Polymyxin B sulfate in sulfuric acid Polymyxin B sulfate bulk drug or preparation, or be further used for monitoring the more slime moulds of sulfuric acid
Plain B, its bulk drug or preparation preparation technology, it is practical.
Brief description of the drawings
Fig. 1 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate in embodiment 1；
Fig. 2~Fig. 5 is concentration-peak area double-log matched curve figure of sulfuric acid Polymyxin B sulfate each component in embodiment 2；
Fig. 6 is the HPLC figures of each component content detection in sulfuric acid Polymyxin B sulfate bulk drug in embodiment 3；
Fig. 7 is the HPLC figures of each component content detection in injection sulfuric acid Polymyxin B sulfate in embodiment 4；
Fig. 8 is the HPLC figures of each component content detection in injection sulfuric acid Polymyxin B sulfate in embodiment 5；
Fig. 9 is the HPLC figures of each component content detection in injection sulfuric acid Polymyxin B sulfate in embodiment 6；
Figure 10 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate in comparative example 1；
Figure 11 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate in comparative example 2；
Figure 12 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate in comparative example 3；
Figure 13 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate bulk drug in comparative example 4；
Figure 14 is the HPLC figures of the analyzing detecting method of sulfuric acid Polymyxin B sulfate bulk drug in comparative example 5；
Embodiment
The present invention is further illustrated below by the mode of embodiment, but does not therefore limit the present invention to described reality
Apply among a scope.The experimental method of unreceipted actual conditions in the following example, conventionally and condition, or according to business
Product specification selects.
Embodiment 1
The specificity of analysis method and sensitivity
Experiment condition：
Trifluoroacetic acid (HPLC levels, U.S. world Co., Ltd)；
Acetonitrile (HPLC levels, Thermo Fisher Scientific Inc.)；
Sulfuric acid Polymyxin B sulfate USP reference substances (contain B in lot number N1M425, every 100mg1 69.7mg、B2 15.9mg、B3
3.2mg、B1-I 6.1mg)；
High performance liquid chromatograph：Alliance e2695 high performance liquid chromatographs (Waters)；
EISD：2424 type EISDs (Waters)；
The high performance liquid chromatograph data handling systems of Empower 3；
Chromatographic column：Kromasil 100-5 C18 chromatographic columns (5 μm, 4.6 × 250mm)；
Mobile phase A is 0.1% trifluoroacetic acid aqueous solution (pH value 1.95), and Mobile phase B is acetonitrile, the ladder of acetonitrile volume ratio
Degree change turns to 0min to 5min, is maintained at 24%；5min to 8min, 29% is risen to from 24%；8min to 15min, from 29% liter
To 32%；15min to 20min, 40% is risen to from 32%.Flow velocity 1.0mL/min, 35 DEG C of column temperature.
60 DEG C of detector drift tube temperature, 12 DEG C, nebulizer gas pressure 35.0psi of atomization mouth temperature, gain 800.
Experimental procedure
Precision weighs sulfuric acid Polymyxin B sulfate USP reference substance 50mg, is placed in 100mL volumetric flasks, with water-acetonitrile (80:
20) dissolve for solvent and be diluted to scale.The μ L of sample introduction 20, repeat sample introduction 6 times.Record chromatogram.
Calculate the RSD of the pin peak area logarithm of each component six, sulfuric acid Polymyxin B sulfate2、B3、B1-I、B1RSD be according to this
0.22%th, 0.50%, 0.29% and 0.13%.By taking the spectrogram of the first pin sample introduction as an example, as shown in figure 1, and combining knowable to table 4：
Sulfuric acid Polymyxin B sulfate2、B3、B1-I、B1Retention time be followed successively by 10.904min, 11.230min, 11.854min,
12.409min, it is kept completely separate between four components, and other impurities are noiseless.As a result show that system is accurate good, this method is to sulfuric acid
The each component of Polymyxin B sulfate has good specificity.
Table 4
| Ingredient names | Retention time/min | Peak area | Separating degree |
| Polymyxin B sulfate2 | 10.904 | 521591 | - |
| Polymyxin B sulfate3 | 11.230 | 38276 | 1.9 |
| Polymyxin B sulfate1-I | 11.854 | 101355 | 3.8 |
| Polymyxin B sulfate1 | 12.409 | 5242581 | 3.1 |
In addition, take identical reference substance solution, using method sample detection under Polymyxin B sulfate component check item in USP40,
As a result display removes Polymyxin B sulfate1Outside, each component response is respectively less than 50mAU.By this method and USP40 analyzing detecting method
Signal to noise ratio and theoretical cam curve be compared, as a result as shown in table 5：
Table 5
As a result show, in this method the detection sensitivity for the sulfuric acid Polymyxin B sulfate each component that relative amount differs greatly and
Post effect has obtained significant raising, is advantageous to improve the integration degree of accuracy of chromatographic peak area.
Embodiment 2
The linear and scope of analysis method
Experiment condition
Tested using experiment condition similar to Example 1.Sulfuric acid Polymyxin B sulfate USP reference substance (lot numbers
Contain B in N1M425, every 100mg1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg)。
Experimental procedure
Precision weighs sulfuric acid Polymyxin B sulfate USP reference substance 100mg, is placed in 20mL volumetric flasks, with water-acetonitrile (80:
20) dissolve and be diluted to scale for solvent, draw the mother liquor 600 μ L, 800 μ L, 900 μ L, 1000 μ L, 1100 μ L, 1200 μ L,
1400 μ L, 2000 μ L and 3000 μ L, are respectively placed in 10mL volumetric flasks, and comparison liquid is used as after diluting constant volume.Take above-mentioned solution each
20 μ L sample introductions, record chromatogram.Using each component peak area take 10 for bottom logarithm as abscissa, it is dense with the quality of respective components
10 logarithms for being bottom that take of degree are ordinate, are fitted conic section.
It is computed, in the range of concentration 0.0457mg/mL~0.165mg/mL, Polymyxin B sulfate2Regression equation be y=-
3.4464x2- 4.7041x+4.7527, R2=0.9978；In the range of concentration 0.0092mg/mL~0.0499mg/mL, more slime moulds
Plain B3Regression equation be y=-1.8517x2- 3.5x+4.3049, R2=0.9959；Concentration 0.0175mg/mL~
In the range of 0.0951mg/mL, Polymyxin B sulfate1-I regression equation is y=-2.2471x2- 3.5297x+4.8556, R2=
0.9964；In the range of concentration 0.2003mg/mL~0.4673mg/mL, Polymyxin B sulfate1Regression equation be y=-2.773x2–
0.0979x+7.3827, R2=0.9980.As shown in Fig. 2~Fig. 5.
As a result show, this method is for Polymyxin B sulfate each component log concentration-peak area logarithm as defined in pharmacopoeia of each country
Quality control clearance in have it is good linear.
Embodiment 3
The content detection of each component in sulfuric acid Polymyxin B sulfate bulk drug
Experiment condition
Tested using experiment condition similar to Example 1.It the difference is that only：The gradient of acetonitrile volume ratio becomes
0min to 5min is turned to, is maintained at 24%；5min to 8min, 27% is risen to from 24%；8min to 15min, risen to from 27%
30%；15min to 20min, 40% is risen to from 30%.Nebulizer gas pressure is 30psi.Batch of sulfuric acid Polymyxin B sulfate USP reference substances
Number be N1M425, per 100mg in contain B1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg.Sulfuric acid Polymyxin B sulfate is former
The lot number for expecting medicine is A1411105, purchased from Xellia drugmakers of Denmark.
Experimental procedure
Precision weighs sulfuric acid Polymyxin B sulfate USP reference substance 50mg, is placed in 10mL volumetric flasks, with water-acetonitrile (80:20)
Dissolved for solvent and be diluted to scale, draw the mother liquor 600 μ L, 900 μ L, 1000 μ L, 1100 μ L, 1400 μ L, 2000 μ L and
3000 μ L, are respectively placed in 10mL volumetric flasks, and comparison liquid is used as after diluting constant volume.Precision weighs sulfuric acid Polymyxin B sulfate 50mg,
It is placed in 100Ml volumetric flasks, with water-acetonitrile (80:20) dissolve for solvent and be diluted to scale, as need testing solution.Take
Each 20 μ L sample introductions of solution are stated, record chromatogram.Using comparison liquid each component peak area take 10 for bottom logarithm as abscissa, with phase
10 logarithms for being bottom that take for answering the mass concentration of component are ordinate, are fitted conic section.In test sample the content of each component by
The logarithm of the peak area measured substitutes into standard curve and calculated.Sulfuric acid Polymyxin B sulfate each component content sum is total content.
As shown in fig. 6, Polymyxin B sulfate each component separating degree is good, Polymyxin B sulfate2Peak and B3The separating degree at peak is 2.1, more
Colistin B3Peak and B1The separating degree at-I peaks is 3.9, Polymyxin B sulfate1- I peaks and B1The separating degree at peak is 2.8.It is computed, it is more glutinous
Rhzomorph B2、B3、B1-I、B1Content be respectively 64.4%, 11.2%, 3.7% and 9.2%, the content of Polymyxin B sulfate is
88.5%.
Embodiment 4
The content detection of injection sulfuric acid Polymyxin B sulfate
Experiment condition
Tested using experiment condition and step similar to Example 3, wherein the aqueous phase in mobile phase is replaced by body
The trifluoroacetic acid aqueous solution that product percentage composition is 0.1%.It is 30 DEG C to be atomized mouth temperature, and drift tube temperature is 70 DEG C.Injection sulphur
Sour Polymyxin B sulfate is makes by oneself, lot number 1507801.
As shown in fig. 7, Polymyxin B sulfate each component separating degree is good, Polymyxin B sulfate2Peak and B3The separating degree at peak is 1.8, more
Colistin B3Peak and B1The separating degree at-I peaks is 3.8, Polymyxin B sulfate1- I peaks and B1The separating degree at peak is 3.2.It is computed, it is more glutinous
Rhzomorph B2、B3、B1-I、B1Content be respectively 64.5%, 11.0%, 3.4% and 7.3%, the content of Polymyxin B sulfate is
86.2%.
Embodiment 5
Tested using experiment condition and step similar to Example 1, wherein the aqueous phase in mobile phase is replaced by body
The trifluoroacetic acid aqueous solution that product percentage composition is 0.01%.The lot number of sulfuric acid Polymyxin B sulfate USP reference substances is N1M425, often
Contain B in 100mg1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg。
As shown in figure 8, Polymyxin B sulfate each component separating degree is good, Polymyxin B sulfate2Peak and B3The separating degree at peak is 2.1, more
Colistin B3Peak and B1The separating degree at-I peaks is 3.1, Polymyxin B sulfate1- I peaks and B1The separating degree at peak is 2.0.With Polymyxin B sulfate1
Peak meter, theoretical cam curve 18243, signal to noise ratio S/N are 1149.
Embodiment 6
Tested using experiment condition and step similar to Example 1, wherein the aqueous phase in mobile phase is replaced by body
The trifluoroacetic acid aqueous solution that product percentage composition is 0.01%.The lot number of sulfuric acid Polymyxin B sulfate USP reference substances is N1M425, often
Contain B in 100mg1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg。
As shown in figure 9, Polymyxin B sulfate each component separating degree is good, Polymyxin B sulfate2Peak and B3The separating degree at peak is 2.1, more
Colistin B3Peak and B1The separating degree at-I peaks is 3.1, Polymyxin B sulfate1- I peaks and B1The separating degree at peak is 2.0.With Polymyxin B sulfate1
Peak meter, theoretical cam curve 18243, signal to noise ratio S/N are 1149.
Comparative example 1
Tested using experiment condition and step similar to Example 1, wherein the aqueous phase in mobile phase is replaced by body
The trifluoroacetic acid aqueous solution that product percentage composition is 0.01%.The lot number of sulfuric acid Polymyxin B sulfate USP reference substances is N1M425, often
Contain B in 100mg1 69.7mg、B2 15.9mg、B3 3.2mg、B1-I 6.1mg。
As shown in Figure 10, Polymyxin B sulfate each component separating degree is good, Polymyxin B sulfate2Peak and B3The separating degree at peak is 2.1,
Polymyxin B sulfate3Peak and B1The separating degree at-I peaks is 3.1, Polymyxin B sulfate1- I peaks and B1The separating degree at peak is 2.0.With polymyxin
B1Peak meter, theoretical cam curve 18243, signal to noise ratio S/N are 1149.
Comparative example 2
Reference implementation example 1, ammonium formate (pH 2.1) aqueous solution that aqueous phase is 20mmol/L is the difference is that only, can not incited somebody to action
Polymyxin B sulfate1- I and B1Separate.As shown in figure 11, each component retains very weak, and can not separate.
Comparative example 3
Reference implementation example 1, the difference is that only aqueous phase is 0.1% formic acid, can not separate each component of Polymyxin B sulfate
Open.As shown in figure 12, each component retains very weak, and can not separate.
Comparative example 4
In the comparative example, containing for sulfuric acid Polymyxin B sulfate is carried out using experiment condition similar to Example 3 and step
Amount detection, the difference is that only aqueous phase is 0.1% formic acid；Use Kromasil C18 (4.6 × 250mm, 5 μm)；Acetonitrile body
The graded of product ratio is 0min to 5min, is maintained at 12%；5min to 8min, 20% is risen to from 12%；8min to 15min,
30% is risen to from 20%；15min to 20min, 40% is risen to from 30%.；The drift tube temperature for the detector that evaporative light-scattering uses
Spend for 70 DEG C；Flow rate of mobile phase 0.8ml/min；The drift tube temperature for the detector that evaporative light-scattering uses is 40 DEG C.
As shown in figure 13, can not be by Polymyxin B sulfate under the present embodiment1- I and B1Separation.
Comparative example 5
In the present embodiment, the content that sulfuric acid Polymyxin B sulfate is carried out using experiment condition similar to Example 3 and step is examined
Survey, the difference is that only aqueous phase is 20mM ammonium acetate aqueous solutions (pH 4.6)；The graded of acetonitrile volume ratio be 0min extremely
5min, it is maintained at 15%；5min to 8min, 24% is risen to from 15%；8min to 15min, 32% is risen to from 24%；15min is extremely
20min, 40% is risen to from 32%；The drift tube temperature for the detector that evaporative light-scattering uses is 70 DEG C；Nebulizer gas pressure is
35psi。
As shown in figure 14, each component of Polymyxin B sulfate can not be separated under the present embodiment.
Claims (10)
1. a kind of analyzing detecting method of sulfuric acid Polymyxin B sulfate, it is characterised in that comprise the steps of：Using high-efficient liquid phase color
Spectrometry combines evaporative light-scattering device, and analysis detection is carried out to sulfuric acid Polymyxin B sulfate, you can；
Chromatographic column in described high performance liquid chromatography is inverse analysis post, and the chromatographic column fixed phase is bonded for octadecyl
Silica gel；
Mobile phase in described high performance liquid chromatography is polar organic solvent and trifluoroacetic acid aqueous solution, the trifluoroacetic acid
The volumn concentration of trifluoroacetic acid is 0.01%~0.3% in the aqueous solution.
2. analyzing detecting method as claimed in claim 1, it is characterised in that described high performance liquid chromatography is washed using gradient
De-, the volume ratio of the trifluoroacetic acid aqueous solution and the polar organic solvent is 85 during the gradient elution:15~60:40.
3. analyzing detecting method as claimed in claim 2, it is characterised in that in the gradient elution, the mobile phase ratio
The scope that changes with time is as shown in table 1：
Table 1
4. the analyzing detecting method described in Claims 2 or 3, it is characterised in that in the gradient elution, the mobile phase ratio
The scope that changes with time is as shown in table 2 or table 3：
Table 2
Table 3
5. analyzing detecting method as claimed in claim 1, it is characterised in that the trifluoroacetic acid in the trifluoroacetic acid aqueous solution
Volumn concentration be 0.05%~0.1%.
6. analyzing detecting method as claimed in claim 1, it is characterised in that the polar organic solvent is selected from methanol, ethanol
With the one or more in acetonitrile；
And/or the model Alliance e2695 that the high performance liquid chromatograph in the high performance liquid chromatography is Waters
High performance liquid chromatograph；
And/or the chromatographic column is Kromasil 100-5C18 posts；
And/or the filler particles degree of the chromatographic column is 2.7 μm~5 μm；It is preferred that 3 μm~5 μm；The length of the chromatographic column is
100mm~250mm, preferably 250mm；
And/or the sampling volume of the chromatographic column is 1~50 μ L, preferably 20 μ L；
And/or the column temperature in the high performance liquid chromatography is 20~40 DEG C, preferably 35 DEG C；
And/or the flow velocity of described mobile phase is 0.5~1.5mL/min, preferably 0.8~1.0mL/min.
7. analyzing detecting method as claimed in claim 1, it is characterised in that the evaporative light-scattering device is that producer is Waters
Model 2424 evaporative light-scattering device；
And/or the drift tube temperature of the evaporative light-scattering device is 40~80 DEG C, preferably 60~70 DEG C；
And/or the nebulizer gas pressure of the evaporative light-scattering device is 20~45psi, preferably 25~35psi；
And/or the atomization mouth temperature of the evaporative light-scattering device is 12 DEG C~35 DEG C；It is preferred that 12~30 DEG C.
8. a kind of analyzing detecting method as described in claim any one of 1-7 contains in sulfuric acid Polymyxin B sulfate bulk drug or preparation
Application in amount detection.
9. application as claimed in claim 9, it is characterised in that the preparation is injection sterile powder.
10. application as claimed in claim 8, it is characterised in that comprise the steps of：Step (1), finite concentration is respectively configured
Reference substance solution and sulfuric acid Polymyxin B sulfate bulk drug or preparation product to be tested solution, using above-mentioned analyzing detecting method to described
Reference substance solution and the sulfuric acid Polymyxin B sulfate preparation product to be tested solution carry out analysis detection；Measure the reference substance solution and
The peak area of each component in the sulfuric acid Polymyxin B sulfate preparation；
Step (2), the peak area of each component and corresponding component in sulfuric acid Polymyxin B sulfate according to the reference substance solution
Concentration, obtain regression equation；It is preferred that the regression equation calculates acquisition by being fitted conic section；
The content of each component of step (3), the sulfuric acid Polymyxin B sulfate bulk drug or preparation is by each group that is measured in step (1)
The regression equation obtained in the substitution step (2) of the peak area divided, you can.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108061767A (en) * | 2017-12-06 | 2018-05-22 | 重庆华邦制药有限公司 | The method of HPLC method separation determination Rivaroxaban intermediates and its related impurities |
| CN108693293A (en) * | 2018-07-25 | 2018-10-23 | 广州白云山医药集团股份有限公司白云山制药总厂 | The method for detecting impurity in amoxicillin granules |
| CN109030610A (en) * | 2018-09-28 | 2018-12-18 | 上海市食品药品检验所 | It is a kind of for measuring the capillary electrophoresis method and its running buffer of polymyxin class antibacterial peptide |
| CN111072757A (en) * | 2020-01-02 | 2020-04-28 | 上海上药第一生化药业有限公司 | Polymyxin B component or salt thereof, and preparation method and application thereof |
| CN111505142A (en) * | 2020-04-26 | 2020-08-07 | 上海市食品药品检验所 | Polymyxin B amino acid configuration analysis method and N-polypeptide terminal sequence sequencing method |
| CN111665307A (en) * | 2020-07-14 | 2020-09-15 | 南京品生医疗科技有限公司 | Kit for detecting concentrations of polymyxin B1and polymyxin B2 in serum |
| CN111830153A (en) * | 2020-07-14 | 2020-10-27 | 南京品生医学检验实验室有限公司 | Method for detecting concentrations of polymyxin B1and polymyxin B2 in serum |
| US11440940B2 (en) | 2020-01-02 | 2022-09-13 | Sph No. 1 Biochemical & Pharmaceutical Co., Ltd. | Polymyxin B component or salt thereof, and preparation and application thereof |
| CN115128191A (en) * | 2022-07-18 | 2022-09-30 | 镇江威特药业有限责任公司 | Method for detecting colistin sulfate content |
| CN115629141A (en) * | 2022-11-02 | 2023-01-20 | 江苏联环药业股份有限公司 | Method for determining content of colistin sulfate tablets by high performance liquid chromatography |
| CN115746101A (en) * | 2022-12-06 | 2023-03-07 | 无锡福祈制药有限公司 | Preparation method of polymyxin B2 |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103130876A (en) * | 2011-11-30 | 2013-06-05 | 天津市海德安科医药科技发展有限公司 | Preparing method of high-purity polymyxin B |
-
2017
- 2017-08-03 CN CN201710656465.6A patent/CN107367562B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103130876A (en) * | 2011-11-30 | 2013-06-05 | 天津市海德安科医药科技发展有限公司 | Preparing method of high-purity polymyxin B |
Non-Patent Citations (3)
| Title |
|---|
| CINDY GOVAERTS等: "Characterization of Polypeptide Antibiotics of the Polymyxin Series by Liquid Chromatography Electrospray Ionization Ion Trap Tandem Mass Spectrometry", 《JOURNAL OF PEPTIDE SCIENCE》 * |
| KIM H. HEE等: "Development and validation of liquid chromatography tandem mass spectrometry method quantitative determination of polymyxin B1,polymyxin B2, polymyxin B3 and isoleucine-polymyxin B1 in human plasma and its application in clinical studies", 《JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS》 * |
| 刘孟娟等: "液质联用技术检测硫酸多黏菌素 B 中各组分", 《中国药业》 * |
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| CN111505142A (en) * | 2020-04-26 | 2020-08-07 | 上海市食品药品检验所 | Polymyxin B amino acid configuration analysis method and N-polypeptide terminal sequence sequencing method |
| CN111830153A (en) * | 2020-07-14 | 2020-10-27 | 南京品生医学检验实验室有限公司 | Method for detecting concentrations of polymyxin B1and polymyxin B2 in serum |
| CN111665307A (en) * | 2020-07-14 | 2020-09-15 | 南京品生医疗科技有限公司 | Kit for detecting concentrations of polymyxin B1and polymyxin B2 in serum |
| CN115128191A (en) * | 2022-07-18 | 2022-09-30 | 镇江威特药业有限责任公司 | Method for detecting colistin sulfate content |
| CN115629141A (en) * | 2022-11-02 | 2023-01-20 | 江苏联环药业股份有限公司 | Method for determining content of colistin sulfate tablets by high performance liquid chromatography |
| CN115746101A (en) * | 2022-12-06 | 2023-03-07 | 无锡福祈制药有限公司 | Preparation method of polymyxin B2 |
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