CN111380971A - HPLC analysis method of bortezomib related substances - Google Patents
HPLC analysis method of bortezomib related substances Download PDFInfo
- Publication number
- CN111380971A CN111380971A CN201811640847.0A CN201811640847A CN111380971A CN 111380971 A CN111380971 A CN 111380971A CN 201811640847 A CN201811640847 A CN 201811640847A CN 111380971 A CN111380971 A CN 111380971A
- Authority
- CN
- China
- Prior art keywords
- scr
- bortezomib
- derivatization
- benzoyl chloride
- organic solvent
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- GXJABQQUPOEUTA-RDJZCZTQSA-N bortezomib Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)B(O)O)NC(=O)C=1N=CC=NC=1)C1=CC=CC=C1 GXJABQQUPOEUTA-RDJZCZTQSA-N 0.000 title claims abstract description 22
- 229960001467 bortezomib Drugs 0.000 title claims abstract description 22
- 239000000126 substance Substances 0.000 title claims abstract description 21
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000004128 high performance liquid chromatography Methods 0.000 title claims abstract description 13
- 238000001212 derivatisation Methods 0.000 claims abstract description 21
- 239000000243 solution Substances 0.000 claims abstract description 21
- PASDCCFISLVPSO-UHFFFAOYSA-N benzoyl chloride Chemical compound ClC(=O)C1=CC=CC=C1 PASDCCFISLVPSO-UHFFFAOYSA-N 0.000 claims abstract description 17
- 239000003960 organic solvent Substances 0.000 claims abstract description 11
- 239000000337 buffer salt Substances 0.000 claims abstract description 7
- 229910019142 PO4 Inorganic materials 0.000 claims abstract description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 claims abstract description 5
- 239000010452 phosphate Substances 0.000 claims abstract description 5
- 239000007788 liquid Substances 0.000 claims abstract description 4
- 239000011259 mixed solution Substances 0.000 claims abstract description 3
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 52
- 238000001514 detection method Methods 0.000 claims description 19
- 238000004458 analytical method Methods 0.000 claims description 13
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 10
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 5
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 5
- 239000003054 catalyst Substances 0.000 claims description 4
- 239000007810 chemical reaction solvent Substances 0.000 claims description 4
- 239000008055 phosphate buffer solution Substances 0.000 claims description 3
- 239000011248 coating agent Substances 0.000 claims description 2
- 238000000576 coating method Methods 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
- 150000004676 glycans Chemical class 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
- 229920001282 polysaccharide Polymers 0.000 claims description 2
- 239000005017 polysaccharide Substances 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
- BTBUEUYNUDRHOZ-UHFFFAOYSA-N Borate Chemical compound [O-]B([O-])[O-] BTBUEUYNUDRHOZ-UHFFFAOYSA-N 0.000 claims 1
- 238000003556 assay Methods 0.000 claims 1
- 150000001875 compounds Chemical class 0.000 claims 1
- 239000012535 impurity Substances 0.000 abstract description 22
- 238000000926 separation method Methods 0.000 abstract description 8
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 30
- GQHTUMJGOHRCHB-UHFFFAOYSA-N 2,3,4,6,7,8,9,10-octahydropyrimido[1,2-a]azepine Chemical group C1CCCCN2CCCN=C21 GQHTUMJGOHRCHB-UHFFFAOYSA-N 0.000 description 20
- 238000005303 weighing Methods 0.000 description 15
- 239000000523 sample Substances 0.000 description 11
- 238000001816 cooling Methods 0.000 description 10
- 238000007865 diluting Methods 0.000 description 10
- 239000000706 filtrate Substances 0.000 description 10
- 238000001914 filtration Methods 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 10
- 239000011550 stock solution Substances 0.000 description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 10
- 239000000047 product Substances 0.000 description 7
- 238000004090 dissolution Methods 0.000 description 6
- SGRPQFBFOBJKQV-UHFFFAOYSA-N acetonitrile;benzoyl chloride Chemical compound CC#N.ClC(=O)C1=CC=CC=C1 SGRPQFBFOBJKQV-UHFFFAOYSA-N 0.000 description 5
- 239000013558 reference substance Substances 0.000 description 5
- 239000012488 sample solution Substances 0.000 description 5
- 239000012085 test solution Substances 0.000 description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- 239000012490 blank solution Substances 0.000 description 2
- 239000004327 boric acid Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- CLBOGHXYEDWPPA-UHFFFAOYSA-N FC(C(=O)O)(F)F.CO.C(C)O Chemical compound FC(C(=O)O)(F)F.CO.C(C)O CLBOGHXYEDWPPA-UHFFFAOYSA-N 0.000 description 1
- DTQVDTLACAAQTR-UHFFFAOYSA-M Trifluoroacetate Chemical compound [O-]C(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-M 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 150000001263 acyl chlorides Chemical class 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 239000002246 antineoplastic agent Substances 0.000 description 1
- 229940041181 antineoplastic drug Drugs 0.000 description 1
- KGBXLFKZBHKPEV-UHFFFAOYSA-N boric acid Chemical compound OB(O)O KGBXLFKZBHKPEV-UHFFFAOYSA-N 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 1
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 1
- 238000002309 gasification Methods 0.000 description 1
- PJXFRXZCERRQPM-UHFFFAOYSA-N hexane;2,2,2-trifluoroacetic acid Chemical compound CCCCCC.OC(=O)C(F)(F)F PJXFRXZCERRQPM-UHFFFAOYSA-N 0.000 description 1
- 239000002953 phosphate buffered saline Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000007974 sodium acetate buffer Substances 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
Images
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
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
-
- 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
- G01N30/62—Detectors specially adapted therefor
- G01N30/74—Optical detectors
-
- 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
- G01N30/04—Preparation or injection of sample to be analysed
- G01N30/06—Preparation
- G01N2030/067—Preparation by reaction, e.g. derivatising the sample
Landscapes
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
The invention discloses an HPLC analysis method of bortezomib related substances, which comprises the following steps: carrying out derivatization treatment on related substances of bortezomib by using benzoyl chloride to obtain a derivatization product; a reversed-phase chiral chromatographic column is adopted, a mobile phase is a mixed solution of a buffer salt solution containing phosphate and an organic solvent, and a high performance liquid chromatography-ultraviolet detector is adopted to detect the derivatization product. The derivatization operation is simple, the derivatization product is single, and the separation degree is good, so that the SCR-3081 and isomer impurities SCR-5220 and SCR-5221 thereof can be simply, rapidly and stably detected, and the product quality can be effectively controlled.
Description
Technical Field
The invention belongs to the technical field of drug analysis, and particularly relates to an HPLC (high performance liquid chromatography) analysis method for related substances of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate.
Background
(1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081) is a raw material for synthesizing bortezomib and is also an important medical intermediate, and the structural formula is shown as the following formula I:
the (1R) - (S) -pinanediol-1-trifluoroacetate ammonium-3-methylbutane-1-boronic acid ester (SCR-3081) typically presents 2 larger isomeric impurities: SCR-5220(SCR-3081 enantiomer) and SCR-5221(SCR-3081 diastereomer). Wherein the structural formula of the SCR-5220 is shown as the following formula II, and the structural formula of the SCR-5221 is shown as the following formula III:
since the isomer impurities of SCR-3081 continuously participate in the reaction in the subsequent steps to form a plurality of corresponding impurities, thereby affecting the quality of the anti-tumor drug bortezomib, it is necessary to establish an analysis method of SCR-3081, control the content of the isomer impurities, improve the purity of the target product, and further improve the quality of the bortezomib drug.
Meanwhile, as the (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081) exists in the form of trifluoroacetate, gasification is difficult, and ultraviolet absorption of SCR-3081 and isomer impurities SCR-5220 and SCR-5221 of the SCR-3081 is weak, a common HPLC analysis method is adopted, and the method has poor separation degree, low response and larger error.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide an HPLC analysis method of bortezomib related substances, which can quickly and accurately realize qualitative, quantitative and isomer analysis.
The invention discloses an HPLC analysis method of bortezomib related substances, which is characterized by comprising the following steps:
(1) carrying out derivatization treatment on related substances of bortezomib by using benzoyl chloride to obtain a derivatization product;
(2) a reversed-phase chiral chromatographic column is adopted, a mobile phase is a mixed solution of a buffer salt solution containing phosphate and an organic solvent, and a high performance liquid chromatography-ultraviolet detector is adopted to detect the derivatization product.
Wherein in the step, the bortezomib related substances are mainly selected from the following structures:
the derivatization treatment in the step (1) is that related substances of bortezomib react with benzoyl chloride in the presence of a catalyst and a reaction solvent to obtain a derivatization product;
further, the catalyst is DBU; the reaction solvent is one or two selected from tetrahydrofuran, acetonitrile and chloroform.
Wherein the derivatized product is selected from the group consisting essentially of the following structures:
the dosage of the organic solvent in the derivatization treatment process is 15-500 mL, preferably 15-300 mL, and more preferably 30-150 mL per 100mg of the organic solvent used by the related substances of the boric acid zomib.
The reaction temperature in the derivatization treatment process is 25-65 ℃, and preferably 45-65 ℃.
The volume of benzoyl chloride used by each 100mg of bortezomib related substances in the derivatization treatment process is 10-200 mul, preferably 10-80 mul, and further preferably 10-50 mul.
In the derivatization treatment process, the volume of DBU used per 100mg of bortezomib is 50-500 mul, and more preferably 100-500 mul.
The reversed-phase chiral chromatographic column in the step 2 is a polysaccharide derivative normal-phase coating type chiral chromatographic column.
The buffer salt in the phosphate-containing buffer salt solution in the step 2 is selected from one or more of disodium hydrogen phosphate, dipotassium hydrogen phosphate, sodium dihydrogen phosphate and potassium dihydrogen phosphate.
The pH value of the phosphate buffer solution in the step 2 is 2.0-7.0, and preferably 3.0-6.0.
The organic solvent in the derivatization treatment process is one or two of tetrahydrofuran, acetonitrile and chloroform.
The organic solvent in the step 2 is one or two of tetrahydrofuran, acetonitrile and chloroform.
The volume proportion of the organic solvent in the mobile phase in the step 2 is 25-75%, preferably 30-60%, and more preferably 35-55%.
When the high performance liquid chromatography-ultraviolet detector in the step (2) is used for detection, the column temperature is 15-50 ℃, and the preferred column temperature is
20-30 ℃; the flow rate of the mobile phase is 0.1-1.0 ml/min, and the preferred flow rate is 0.4-0.6 ml/min; detection of ultraviolet detector
The detection wavelength is 220-260 nm, and the preferred detection wavelength is 230-240 nm.
The invention provides an analysis method of bortezomib related substances, which is characterized in that the analysis is carried out on the derivatized bortezomib related substances by utilizing high performance liquid chromatography, the derivatization operation is simple, the derivatization product is single, and the separation degree is good, so that SCR-3081 and isomer impurities SCR-5220 and SCR-5221 thereof can be simply, rapidly and stably detected, and the product quality can be effectively controlled.
Drawings
FIG. 1: HPLC detection blank solution chromatogram in example 1
FIG. 2: HPLC detection System suitability Profile in example 1
FIG. 3: HPLC detection profile of SCR-3081 in example 1
Detailed Description
The invention is further explained below by way of examples, which should not be construed as limiting the scope of the invention.
Example 1:
1) chromatographic conditions are as follows:
the instrument comprises the following steps: agilent1260HPLC
Mobile phase: 10mmol/L of K2HPO4Buffer (pH adjusted to 5.0 with phosphoric acid) -acetonitrile (55: 45 by volume)
A chromatographic column:
OX-3(4.6×150mm,3μm)
detection wavelength: 235nm
Column temperature: 25 deg.C
Flow rate: 0.4ml/min
Sample introduction amount: 5ul
2) Sample preparation:
taking about 50mg of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081), precisely weighing, placing in a 25ml measuring flask, adding 20 mu l benzoyl chloride, shaking up, adding 50 mu l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), adding 15ml acetonitrile, carrying out ultrasonic dissolution, reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution.
Accurately weighing SCR-5220 and SCR-5221 reference substances respectively at 5mg, placing in a 25ml measuring flask, adding 15ml benzoyl chloride-acetonitrile solution (20 μ l benzoyl chloride is placed in a 200ml measuring flask, adding acetonitrile to dilute to scale, shaking up) and ultrasonically dissolving, adding 50 μ l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to scale, shaking up, filtering, and taking the subsequent filtrate as SCR-5220 and SCR-5221 impurity stock solutions respectively.
Accurately weighing an appropriate amount of SCR-3081, accurately adding an appropriate amount of SCR-5220 and SCR-5221 impurity stock solutions respectively according to the preparation method operation of a test solution to prepare solutions containing 2mg, 4 mug and 4 mug of SCR-3081, SCR-5220 and SCR-5221 respectively per 1ml as system applicability solutions.
3) And (3) measuring results: the blank solution chromatogram is shown in figure 1, the system applicability chromatogram is shown in figure 2, and the test sample detection chromatogram is shown in figure 3. The detection result shows that the separation degree of the SCR-3081, the SCR-5220, the SCR-5221 and other impurities is good.
Example 2:
1) chromatographic conditions are as follows:
the instrument comprises the following steps: agilent1260HPLC
Mobile phase: phosphate buffered saline (pH 3.0) -acetonitrile (volume ratio 65:35)
A chromatographic column:
OX-3(4.6×150mm,3μm)
detection wavelength: 235nm
Column temperature: 20 deg.C
Flow rate: 1.0ml/min
Sample introduction amount: 5ul
2) Sample preparation:
taking about 50mg of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-boric acid ester (SCR-3081), precisely weighing, placing in a 25ml measuring flask, adding 20, shaking up benzoyl chloride, adding 50 acyl chloride, shaking up (1,8 diazabicyclo [5,4,0] undec-7-ene), adding 15ml acetonitrile, carrying out ultrasonic dissolution, reacting in a water bath at 60 ℃ for 30min, cooling to room temperature, diluting to a scale with methanol, shaking up, filtering, and taking a subsequent filtrate as a sample solution.
Accurately weighing SCR-5220 and SCR-5221 reference substances respectively at 5mg, placing in a 25ml measuring flask, adding 15ml benzoyl chloride-acetonitrile solution (20 solution benzoyl chloride is placed in a 200ml measuring flask, adding acetonitrile to dilute to scale, shaking up) for ultrasonic dissolution, adding 50ml acetonitrile dilute (1,8 diazabicyclo [5,4,0] undec-7-ene), reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to scale, shaking up, filtering, and taking the subsequent filtrate as SCR-5220 and SCR-5221 impurity stock solutions respectively.
Accurately weighing an appropriate amount of SCR-3081, accurately adding an appropriate amount of SCR-5220 and SCR-5221 impurity stock solutions respectively according to the preparation method operation of a test solution to prepare solutions containing 2mg, 4mg and 4mg of SCR-3081, SCR-5220 and SCR-5221 respectively per 1ml as system applicability solutions.
3) And (3) measuring results: the detection result shows that the separation degree of the SCR-3081, the SCR-5220, the SCR-5221 and other impurities is good.
Example 3:
1) chromatographic conditions are as follows:
the instrument comprises the following steps: agilent1260HPLC
Mobile phase: phosphate buffer solution (pH 6.5) -acetonitrile (40:60)
A chromatographic column:
OX-3(4.6×150mm,3μm)
detection wavelength: 235nm
Column temperature: 30 deg.C
Flow rate: 0.2ml/min
Sample introduction amount: 5ul
2) Sample preparation:
taking about 50mg of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081), precisely weighing, placing in a 25ml measuring flask, adding 20 mu l benzoyl chloride, shaking up, adding 50 mu l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), adding 15ml acetonitrile, carrying out ultrasonic dissolution, reacting in a water bath at 45 ℃ for 30min, cooling to room temperature, diluting with methanol to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution.
Accurately weighing SCR-5220 and SCR-5221 reference substances respectively at 5mg, placing in a 25ml measuring flask, adding 15ml benzoyl chloride-acetonitrile solution (20 μ l benzoyl chloride is placed in a 200ml measuring flask, adding acetonitrile to dilute to scale, shaking up) and ultrasonically dissolving, adding 50 μ l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to scale, shaking up, filtering, and taking the subsequent filtrate as SCR-5220 and SCR-5221 impurity stock solutions respectively.
Accurately weighing an appropriate amount of SCR-3081, accurately adding an appropriate amount of SCR-5220 and SCR-5221 impurity stock solutions respectively according to the preparation method operation of a test solution to prepare solutions containing 2mg, 4 mug and 4 mug of SCR-3081, SCR-5220 and SCR-5221 respectively per 1ml as system applicability solutions.
3) And (3) measuring results: the detection result shows that the separation degree of the SCR-3081, the SCR-5220, the SCR-5221 and other impurities is good.
Example 4:
1) chromatographic conditions are as follows:
the instrument comprises the following steps: agilent1260HPLC
Mobile phase: n-Hexane-trifluoroacetic acid (100: 0.1)/Anhydrous ethanol-methanol-trifluoroacetic acid (75:25:0.1) (volume ratio 90:10)
A chromatographic column:
AY-H(4.6×250mm,5μm)
detection wavelength: 254nm
Column temperature: 35 deg.C
Flow rate: 1.0ml/min
Sample introduction amount: 10ul of
2) Sample preparation:
taking about 50mg of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081), precisely weighing, placing in a 25ml measuring flask, adding 20 mu l benzoyl chloride, shaking up, adding 50 mu l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), adding 15ml acetonitrile, carrying out ultrasonic dissolution, reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution.
Accurately weighing SCR-5220 and SCR-5221 reference substances respectively at 5mg, placing in a 25ml measuring flask, adding 15ml benzoyl chloride-acetonitrile solution (20 μ l benzoyl chloride is placed in a 200ml measuring flask, adding acetonitrile to dilute to scale, shaking up) and ultrasonically dissolving, adding 50 μ l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to scale, shaking up, filtering, and taking the subsequent filtrate as SCR-5220 and SCR-5221 impurity stock solutions respectively.
Accurately weighing an appropriate amount of SCR-3081, accurately adding an appropriate amount of SCR-5220 and SCR-5221 impurity stock solutions respectively according to the preparation method operation of a test solution to prepare solutions containing 2mg, 4 mug and 4 mug of SCR-3081, SCR-5220 and SCR-5221 respectively per 1ml as system applicability solutions.
3) And (3) measuring results: the detection result shows that the separation degree of the SCR-3081, the SCR-5220, the SCR-5221 and other impurities is poor.
Example 5:
1) chromatographic conditions are as follows:
the instrument comprises the following steps: agilent1260HPLC
Mobile phase: sodium acetate buffer (pH 5.0) -acetonitrile (55: 45 by volume)
A chromatographic column:
OX-3(4.6×150mm,3μm)
detection wavelength: 235nm
Column temperature: 25 deg.C
Flow rate: 0.4ml/min
Sample introduction amount: 5ul
2) Sample preparation:
taking about 50mg of (1R) - (S) -pinanediol-1-ammonium trifluoroacetate-3-methylbutane-1-borate (SCR-3081), precisely weighing, placing in a 25ml measuring flask, adding 20 mu l benzoyl chloride, shaking up, adding 50 mu l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), adding 15ml acetonitrile, carrying out ultrasonic dissolution, reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to a scale, shaking up, filtering, and taking a subsequent filtrate as a sample solution.
Accurately weighing SCR-5220 and SCR-5221 reference substances respectively at 5mg, placing in a 25ml measuring flask, adding 15ml benzoyl chloride-acetonitrile solution (20 μ l benzoyl chloride is placed in a 200ml measuring flask, adding acetonitrile to dilute to scale, shaking up) and ultrasonically dissolving, adding 50 μ l DBU (1,8 diazabicyclo [5,4,0] undec-7-ene), reacting in a water bath at 50 ℃ for 30min, cooling to room temperature, diluting with methanol to scale, shaking up, filtering, and taking the subsequent filtrate as SCR-5220 and SCR-5221 impurity stock solutions respectively.
Accurately weighing an appropriate amount of SCR-3081, accurately adding an appropriate amount of SCR-5220 and SCR-5221 impurity stock solutions respectively according to the preparation method operation of a test solution to prepare solutions containing 2mg, 4 mug and 4 mug of SCR-3081, SCR-5220 and SCR-5221 respectively per 1ml as system applicability solutions.
3) And (3) measuring results: the detection result shows that the separation degree of the SCR-3081, the SCR-5220, the SCR-5221 and other impurities is poor.
Claims (12)
1. An HPLC analysis method of bortezomib related substances is characterized by comprising the following steps:
(1) carrying out derivatization treatment on related substances of bortezomib by using benzoyl chloride to obtain a derivatization product;
(2) a reversed-phase chiral chromatographic column is adopted, a mobile phase is a mixed solution of a buffer salt solution containing phosphate and an organic solvent, and a high performance liquid chromatography-ultraviolet detector is adopted to detect the derivatization product.
2. The analytical method according to claim 1, wherein the derivatization treatment in step (1) is a reaction of bortezomib-related substance with benzoyl chloride in the presence of a catalyst and a reaction solvent to obtain a derivatized product; the catalyst is DBU; the reaction solvent is one or two selected from tetrahydrofuran, acetonitrile and chloroform.
3. The analysis method according to claim 2, wherein the amount of the organic solvent is 15 to 500mL, preferably 15 to 300mL, and more preferably 30 to 150mL per 100mg of the zomib borate-related substance.
4. The analytical method according to claim 2, characterized in that the certain temperature is 25 to 65 ℃, preferably 45 to 65 ℃. The reaction temperature of the reaction is 25-65 ℃, and preferably 45-65 ℃.
5. The method according to claim 2, wherein the volume of benzoyl chloride used per 100mg of bortezomib-related substance is 10 to 200. mu.l, preferably 10 to 80. mu.l, and more preferably 10 to 50. mu.l.
6. The method according to claim 2, wherein the volume of DBU used per 100mg of bortezomib-related substance is 50 to 500. mu.l, and more preferably 100 to 500. mu.l.
7. The analytical method according to claim 1, wherein the reverse-phase chiral chromatographic column in step 2 is a polysaccharide derivative normal-phase coating type chiral chromatographic column.
8. The method according to claim 1, wherein the buffer salt in the phosphate-containing buffer salt solution in step 2 is selected from one or more of disodium hydrogenphosphate, dipotassium hydrogenphosphate, sodium dihydrogenphosphate, and potassium dihydrogenphosphate; the pH value of the phosphate buffer solution is 2.0-7.0, preferably 3.0-6.0.
9. The analytical method according to claim 1, wherein the organic solvent is one or two selected from the group consisting of tetrahydrofuran, acetonitrile, and chloroform.
10. The analytical method according to claim 1, wherein the organic solvent in step (2) is in a volume ratio of 25% to 75%, preferably 30% to 60%, and more preferably 35% to 55% of the mobile phase.
11. The analysis method according to claim 1, wherein the column temperature of the HPLC-UV detector in step (2) is 15-50 ℃, preferably 20-30 ℃; the flow rate of the mobile phase is 0.1-1.0 ml/min, and the preferred flow rate is 0.4-0.6 ml/min; the detection wavelength of the ultraviolet detector is 220-260 nm, preferably 230-240 nm.
12. The assay of claim 1, wherein the bortezomib-related substance comprises the following compounds:
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811640847.0A CN111380971B (en) | 2018-12-29 | 2018-12-29 | HPLC analysis method of bortezomib related substances |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811640847.0A CN111380971B (en) | 2018-12-29 | 2018-12-29 | HPLC analysis method of bortezomib related substances |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111380971A true CN111380971A (en) | 2020-07-07 |
| CN111380971B CN111380971B (en) | 2022-04-12 |
Family
ID=71216640
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811640847.0A Active CN111380971B (en) | 2018-12-29 | 2018-12-29 | HPLC analysis method of bortezomib related substances |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111380971B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113402A (en) * | 2021-12-03 | 2022-03-01 | 重庆医药高等专科学校 | Method for determining pinanediol content in bortezomib by adopting high performance liquid chromatography |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103965231A (en) * | 2013-01-31 | 2014-08-06 | 江苏奥赛康药业股份有限公司 | Amide borate for detecting bortezomib intermediate purity, preparation method and application thereof |
| CN104483419A (en) * | 2014-12-31 | 2015-04-01 | 四川汇宇制药有限公司 | Detection method of bortezomib intermediate |
| WO2017019804A2 (en) * | 2015-07-28 | 2017-02-02 | Plexxikon Inc. | Compounds and methods for kinase modulation, and indications therefor |
| CN106706796A (en) * | 2017-01-13 | 2017-05-24 | 南京海辰药业股份有限公司 | Method for detecting optical isomer of key intermediate 1R-trifluoroacetate of bortezomib by HPLC (high performance liquid chromatography) |
| CN106770877A (en) * | 2017-03-29 | 2017-05-31 | 昆明贵研药业有限公司 | A kind of detection method of bortezomib chiral isomer |
| US20170313828A1 (en) * | 2016-05-02 | 2017-11-02 | Virginia Commonwealth University | In situ-forming of dendrimer hydrogels using michael-addition reaction |
-
2018
- 2018-12-29 CN CN201811640847.0A patent/CN111380971B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103965231A (en) * | 2013-01-31 | 2014-08-06 | 江苏奥赛康药业股份有限公司 | Amide borate for detecting bortezomib intermediate purity, preparation method and application thereof |
| CN104483419A (en) * | 2014-12-31 | 2015-04-01 | 四川汇宇制药有限公司 | Detection method of bortezomib intermediate |
| WO2017019804A2 (en) * | 2015-07-28 | 2017-02-02 | Plexxikon Inc. | Compounds and methods for kinase modulation, and indications therefor |
| US20170313828A1 (en) * | 2016-05-02 | 2017-11-02 | Virginia Commonwealth University | In situ-forming of dendrimer hydrogels using michael-addition reaction |
| CN106706796A (en) * | 2017-01-13 | 2017-05-24 | 南京海辰药业股份有限公司 | Method for detecting optical isomer of key intermediate 1R-trifluoroacetate of bortezomib by HPLC (high performance liquid chromatography) |
| CN106770877A (en) * | 2017-03-29 | 2017-05-31 | 昆明贵研药业有限公司 | A kind of detection method of bortezomib chiral isomer |
Non-Patent Citations (2)
| Title |
|---|
| ANDREY S. IVANOV等: "Synthesis and Characterization of Organic Impurities in Bortezomib Anhydride Produced by a Convergent Technology", 《SCIENTIA PHARMACEUTICA》 * |
| 王觉晓等: "HPLC法测定硼替佐米起始合成原料3中的对映异构体杂质", 《中国药师》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114113402A (en) * | 2021-12-03 | 2022-03-01 | 重庆医药高等专科学校 | Method for determining pinanediol content in bortezomib by adopting high performance liquid chromatography |
| CN114113402B (en) * | 2021-12-03 | 2023-05-23 | 重庆医药高等专科学校 | Method for measuring pinanediol content in bortezomib by adopting high performance liquid chromatography |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111380971B (en) | 2022-04-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Jiang et al. | Determination of trace bisphenols in functional beverages through the magnetic solid-phase extraction with MOF-COF composite | |
| CN106706768B (en) | Method for separating and measuring empagliflozin and related substances thereof | |
| CN112697912B (en) | Method for detecting tofacitinib citrate intermediate and related impurities thereof | |
| CN111380971B (en) | HPLC analysis method of bortezomib related substances | |
| CN112480013A (en) | 3, 4-dihydropyrimidine benzonitrile derivative, and preparation method and application thereof | |
| CN108037209B (en) | Liquid chromatography analysis method of ticagrelor chiral intermediate | |
| CN106226426A (en) | A kind of high performance liquid chromatography splits the method for canagliflozin five-membered ring impurity enantiomer | |
| CN109946398B (en) | Method for detecting dalbavancin and impurities thereof | |
| CN106706769B (en) | Separation and determination method of empagliflozin and optical isomer thereof | |
| CN108169399B (en) | Method for separating impurities in ethyl demethylaminothiazolyloximate crude product | |
| CN114184716B (en) | High performance liquid chromatography analysis method for determining related substance components in halominosone cream | |
| CN112213402A (en) | Method for detecting (3R,4R) -1-benzyl-N, 4-dimethylpiperidine-3-amine dihydrochloride and isomer thereof | |
| CN112034066B (en) | Method for separating and measuring Ribociclib and impurities | |
| CN110632197B (en) | Analysis and detection method for benzothiazole and derivatives thereof in dibenzothiazyl disulfide production process | |
| CN114295746A (en) | Analysis and detection method of pitavastatin calcium | |
| CN112213407B (en) | Detection method of levoornidazole related substances | |
| CN112326807B (en) | Method for synthesizing sucrose-6-ethyl ester by central control monitoring monoester method | |
| CN114113357A (en) | Etimicin starting material gentamicin C1aIs detected by | |
| CN113866282A (en) | Separation and detection method for isomer impurities in 3-halogenated-7- (4-bromobenzoyl) -1-hydro-indole and application thereof | |
| CN112666282A (en) | High performance liquid detection method for impurities in calcium zinc gluconate oral solution | |
| CN112834643B (en) | Method for measuring 2, 6-dihydroxy-4-methylpyridine and 2, 6-dihydroxy-3-cyano-4-methylpyridine | |
| CN112595788B (en) | High performance liquid chromatography method for separating (R), (S) -1- (alpha-naphthyl) glycidyl ether | |
| CN111735899B (en) | Analysis method of naphthalene ring containing substituent | |
| CN113702536B (en) | Detection method and application of 6-chloromethyl-2-pyridine methanol | |
| CN109358119B (en) | Detection method of galanthamine hydrobromide intermediate and impurity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230605 Address after: No. 20 Yaogu Yiheng Road, Xiuying District, Haikou City, Hainan Province, 570311 Patentee after: Xiansheng Zaiming Pharmaceutical Co.,Ltd. Address before: 210042 699 Xuanwu Road, Xuanwu District, Nanjing, Jiangsu -18 Patentee before: JIANGSU SIMCERE PHARMACEUTICAL Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| CP03 | Change of name, title or address |
Address after: No. 20 Yaogu Yiheng Road, Xiuying District, Haikou City, Hainan Province, 570311 Patentee after: Hainan Xiansheng Zaiming Pharmaceutical Co.,Ltd. Country or region after: China Address before: No. 20 Yaogu Yiheng Road, Xiuying District, Haikou City, Hainan Province, 570311 Patentee before: Xiansheng Zaiming Pharmaceutical Co.,Ltd. Country or region before: China |
|
| CP03 | Change of name, title or address |