CN103645259A - Method for simultaneously determining 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione and related substances thereof - Google Patents
Method for simultaneously determining 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione and related substances thereof Download PDFInfo
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- CN103645259A CN103645259A CN201310683222.3A CN201310683222A CN103645259A CN 103645259 A CN103645259 A CN 103645259A CN 201310683222 A CN201310683222 A CN 201310683222A CN 103645259 A CN103645259 A CN 103645259A
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Abstract
The invention discloses a method for simultaneously determining 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione and related substances thereof. According to the method, the 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione and the related substances thereof are separated through high performance liquid chromatography, wherein a chromatographic column is bonded by octadecyl silica gel; a mobile phase is a methanol:acetic acid aqueous solution. The flow velocity in the high performance liquid chromatography is 0.5 to 1.2ml/min. The detection wavelength in the high performance liquid chromatography is 226nm. The method is simple, convenient and practical, can be used for quickly achieving the aims of simultaneously determining and separating the 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione and the related substances thereof, can be applied to determination of molecular weights of unknown impurities through liquid chromatography-mass spectrometry (LC-MS), provides a reliable basis for structure inference of the unknown impurities, and facilitates quality control of raw materials and preparations of the 4-amino-2-(2,6-dioxo-3-piperidyl)isoindoline-1,3-dione.
Description
Technical field
The present invention relates to a kind of separation, measure the method for mooring horse degree amine and related substance thereof, especially relate to a kind of method of simultaneously measuring pool horse degree amine, synthetic initiation material, intermediate, accessory substance, degradation product and other unknown impurities.
Background technology
Pool horse degree amination formal name used at school is called 4-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone, is a kind of thalidomide analogs, is a kind of immunomodulator that has antitumor activity.In cytologic experiments, can suppress hematopoiesis tumor cell proliferation and apoptosis-induced in vitro.In addition, pool horse degree amine can suppress the propagation of lenalidomide resistance multiple myeloma cell line and at lenalidomide-sensitivity and lenalidomide-drug-resistant cell strain and dexamethasone co-induction apoptosis of tumor cells.Pool horse degree amine strengthen T cell-and the generation of the cell-mediated immunity of NK cell (NK) and inhibition monocyte pro-inflammatory cytokine (as, TNF-α and IL-6).Mouse tumor model and and external umbilical cord model in moor horse degree amine and show anti-angiogenesis activity, this medicine is in clinical use at present.
Structural formula is:
In this product synthesis technique, with 3-nitrophthalic acid and 3-amino-2,6 ,-piperidine dione is as raw material, and intermediate is 3-aminophthalic acid, may produce the accessory substance of 4 nitros or 5 bit aminos in technological process, and unknown impurity.Adopt the conventional high efficiency liquid phase method of current detection related substance to detect these impurity difficulty larger, in existing known method, be difficult to the complete above known impurities of separation in Same Way, so that there is certain difficulty in the quality control of this product, for safe medication brings relatively large hidden danger.
Summary of the invention
The object of this invention is to provide a kind of method of simultaneously measuring pool horse degree amine and related substance thereof.Through repeatedly optimizing, with efficient liquid-phase chromatography method, reach separated object simultaneously, and can be applicable to the mobile phase of the high efficiency liquid phase method of Mass Spectrometer Method, the convenient structure of determining known and unknown related substance by mass spectrophotometry.
Technical scheme of the present invention is:
Measure a method for pool horse degree amine and related substance thereof simultaneously, comprise the following steps:
(1) preparation system applicability solution:
Get pool horse degree amine and accessory substance 5-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone work reference substance, accurately weighed, adding acetonitrile makes to dissolve in right amount, add mobile phase A: it is all the solution of 0.01mg containing pool horse degree amine, accessory substance that the acetic acid that pH value is 2.6～3.0 is made every 1ml, the final volume of described acetonitrile and mobile phase A acetic acid is than being 20:80 again;
(2) preparation pool horse degree amine need testing solution:
Get pool horse degree amine, accurately weighed, add acetonitrile and make in right amount to dissolve, add mobile phase A: the acetic acid that pH value is 2.6～3.0 is made the solution that every 1ml is 0.125mg containing pool horse degree amine, the final volume of described acetonitrile and mobile phase A acetic acid is than being 20:80;
(3) by following high-efficient liquid phase chromatogram condition, carry out determination of related substances:
A. take octadecylsilane chemically bonded silica as filling agent, the acetum that the pH value of take is 2.6～3.0 is mobile phase A, take methyl alcohol as Mobile phase B, gradient elution, and column temperature is 20～35 ℃, and flow velocity is 0.5～1.2ml/min, and detection wavelength is 226nm;
Described condition of gradient elution is:
B. precision is drawn reference substance solution and need testing solution respectively, and injection liquid chromatography is measured pool horse degree amine and related substance thereof.
Preferably, described in step 3, the pH of acetum is 2.6.
Preferably, described a. be take octadecylsilane chemically bonded silica as 150 * 4.0mm, and 5 μ m filling agents, take acetum as mobile phase A, take first as Mobile phase B, gradient elution, and column temperature is 30 ℃, and flow velocity is 1ml/min, and detection wavelength is 226nm.
Advantage of the present invention is the high performance liquid chromatography by setting up, optimizing, measure pool horse degree amine, synthetic initiation material, intermediate, accessory substance, degradation product and other unknown impurities and related substance thereof simultaneously, a kind of stability, reproducible assay method are provided, can be applied to LC-MS (LC-MS) and determine the molecular weight of unknown impuritie, for inferring that the structure of unknown impuritie provides reliable basis, for mooring the quality control of horse degree amine raw material and preparation, provide reliable basis, can be convenient to this new drug Quality Control.
By above analytical approach, can be effectively that pool horse degree amine and related substance thereof is separated, degree of separation all meets the requirement of chromatographic resolution far away, and saliferous not in mobile phase, noiseless to Mass Spectrometer Method, can be used for Mass Spectra, determine the molecular weight of each impurity peaks, for inferring that unknown impuritie is for reliable foundation.
Below in conjunction with the drawings and specific embodiments, the present invention is described in further detail, according to ordinary skill knowledge and customary means, makes various replacements and change, include within the scope of the invention.
Accompanying drawing explanation
Fig. 1 is that horse degree amine on the berth adds related impurities chromatographic fractionation figure (embodiment 7).
In figure, the chromatographic peak retention time of known raw material, intermediate or accessory substance sees the following form:
Embodiment
Instrument and reagent in following examples are common commercially available except specified otherwise.
Instrument: Agilent1260series high performance liquid chromatograph, chromatographic grade acetonitrile, methyl alcohol (Merck company), glacial acetic acid (analyzing pure).
Chromatographic column: Agilent ZORBAX SB-C185 μ m4.6 * 150mm
Reagent: raw material 1:3-amino-2,6,-piperidine dione (market purchasing), intermediate: 3-aminophthalic acid (market purchasing), raw material 2:3-nitrophthalic acid (market purchasing), accessory substance 1:4 – nitro-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone (self-control), accessory substance 2:5-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone (self-control), pool horse degree amine raw material (Shenzhen Haiwang Pharmaceutical Co., Ltd provides, lot number 20130401,20130402,20130403).
The preparation of embodiment 1 system suitability solution
Get pool horse degree amine and accessory substance 2:5-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone; In right amount, with acetonitrile, make in right amount to dissolve, adding acetic acid that mobile phase A pH value is 2.6～3.0, to be diluted to containing mooring horse degree amine and accessory substance 2 be all the solution of 10 μ g/ml.Acetonitrile and mobile phase A pH value are that the final volume ratio of 2.6～3.0 acetic acid is 20:80.Accessory substance 2 is accessory substances of the most approaching pool horse degree amine, and separated with accessory substance 2, other impurity is separable.
The screening of embodiment 2 chromatographic conditions and system suitability
The selection of 2.1 mobile phases
According to mass detector, measure related request, successively select the separating effect following (in Table 1) under different mobile phase conditions; Acetum is with vinegar acid for adjusting pH in water.
Table 1. mobile phase selection result
| Mobile phase | Separating effect |
| Methyl alcohol-acetum (pH2.2) (30:70) | Degree of separation is 4.3 |
| Methyl alcohol-acetum (pH2.6) (30:70) | Degree of separation is 8.5 |
| Methyl alcohol-acetum (pH3.0) (30:70) | Degree of separation is 6.1 |
| Methyl alcohol-acetum (pH2.6) (gradient 1) | Degree of separation is 11.2 |
| Methyl alcohol-acetum (pH2.6) (gradient 2) | Degree of separation is 9.2 |
According to mooring the degree of separation result (in Table 1) at horse degree amine peak and accessory substance 2 peaks in the measured system suitability solution of different mobile phases, show that gradient elution is than the good separating effect of isocratic elution, the degree of separation that the gradient elution program of employing this law (shown in table 4) obtains increases more obvious.
The selection of 2.2 chromatographic columns
The chromatographic column of different brands on probation, different model, finally determines and adopts Agilent ZORBAXSB-C18(150 * 4.6mm, 5 μ m), chromatographic column selection result is in Table 2.
Table 2. chromatographic column selection result
Adopt the gradient elution program shown in table 4, in different brands C18 chromatographic column, carry out comparison test, result shows (in Table 2), use Agilent ZORBAX SB-C18(150 * 4.6mm, 5 μ m) time, the post effect of pool horse degree amine main peak is higher, and in system suitability solution, the degree of separation of two compositions is better.
Finally determine and take acetum as mobile phase A, take methyl alcohol as Mobile phase B, the optimal pH of acetum is 2.6 left and right, according to following table 4, carries out gradient elution.
The selection of 2.3 column temperatures
Select column temperature to be 25 ℃, 30 ℃ and 35 ℃ and investigate, finally determine that column temperature is 30 ℃, column temperature selection result is in Table 3.
Table 3. column temperature selection result
Adopt the gradient elution program shown in table 4, use Agilent ZORBAX SB-C18(150 * 4.6mm, 5 μ m) chromatographic column, under different column temperatures, to test, result shows (in Table 3), and when column temperature is 30 ℃, degree of separation is better, and main peak post effect is higher.
Determining of chromatographic condition: through above-mentioned selection test, definite chromatographic condition is: with octadecylsilane chemically bonded silica (150 * 4.0mm, 5 μ m) be filling agent, take acetum as mobile phase A, take methyl alcohol as Mobile phase B, gradient elution, column temperature is 30 ℃, flow velocity is 1ml/min, and detection wavelength is 226nm; Described condition of gradient elution is in Table 4.
2.4 system suitabilities:
Chromatographic condition and system suitability: with octadecylsilane chemically bonded silica (Agilent ZORBAXSB-C18150 * 4.6mm, 5 μ m) be filling agent, take acetum as mobile phase A, take methyl alcohol as Mobile phase B, gradient elution, column temperature is 30 ℃, flow velocity is 1ml/min, detection wavelength is 226nm, according to following table 4, carries out gradient 1 wash-out.
Table 4. gradient 1 mobile phase elution program
The preparation of embodiment 3 detectability solution
Detectability solution: precision measures need testing solution 0.11ml, puts in 100ml measuring bottle, adds mobile phase A-Mobile phase B (80:20(v/v)) be diluted to scale, shake up, obtain.
System suitability solution: get pool horse degree amine and accessory substance 2 appropriate, make in right amount to dissolve with acetonitrile, add Mobile phase B and be diluted to containing the solution of mooring horse degree amine and accessory substance 2 and be respectively 10 μ g/ml.
Embodiment 4 detectability tests
Detectability solution 20ul described in accurate absorption embodiment 3, injection liquid chromatography, records chromatogram, calculates main peak signal to noise ratio (S/N ratio) (S/N), 3<(S/N) <5, shows that the detectability concentration of this law mensuration pool horse degree amine is about 0.12 μ g/ml.
The preparation of embodiment 5 specificity solution
Need testing solution: precision takes pool horse degree amine 12.5mg, puts in 100ml measuring bottle, adds acetonitrile 20ml and makes to dissolve, and adds Mobile phase B and is diluted to scale, shakes up, and obtains.
Specificity solution: get pool horse degree amine need testing solution, add in right amount 5 known impurities: raw material 1:3-amino-2,6,-piperidine dione, intermediate: 3-aminophthalic acid, raw material 2:3-nitrophthalic acid, accessory substance 1:4 – nitro-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone, accessory substance 2:5-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone.
Specificity solution 20ul described in accurate absorption embodiment 5, injection liquid chromatography, record chromatogram, the degree of separation of pool horse degree amine main peak and 5 known impurities is all greater than 10, degree of separation between 5 known impurities is all greater than 5, shows that the specificity of this law mensuration pool horse degree amine and 5 known impurities is better.
Embodiment 7 determination of related substances tests
Need testing solution 20ul described in accurate absorption embodiment 5, injection liquid chromatography, records chromatogram.In the pool horse degree amine of three lot numbers (Shenzhen Haiwang Pharmaceutical Co., Ltd provides, lot number 20130401,20130402,20130403), all do not detect 5 known impurities, unknown impuritie total content is respectively 0.03%, 0.04% and 0.02%.
The contrast of embodiment 8 and existing known method
Existing known method chromatographic condition:
Chromatographic column: ACE C18 post 150 * 4.6mm, 3 μ m
Mobile phase A: methyl alcohol
Mobile phase B: 25mM ammonium acetate (add formic acid and adjust pH=5.5)
Both press gradient elution by volume, as following table:
The existing known method mobile phase of table 5. elution program
Flow velocity: 0.7ml/min,
Column temperature: 30 ℃.
By above-mentioned existing method chromatographic condition, precision is drawn the specificity solution 20ul described in embodiment 5, and injection liquid chromatography, records chromatogram, and result and this law are relatively in Table 6
The existing known method of table 6. and this law comparison
Claims (5)
1. measure a method for pool horse degree amine and related substance thereof simultaneously, comprise the following steps:
(1) preparation system applicability solution;
(2) preparation pool horse degree amine need testing solution;
(3) by following high-efficient liquid phase chromatogram condition, carry out determination of related substances: a. and take octadecylsilane chemically bonded silica as filling agent, the acetum that the pH value of take is 2.6～3.0 is mobile phase A, take methyl alcohol as Mobile phase B, gradient elution, column temperature is 20～35 ℃, flow velocity is 0.5～1.2ml/min, and detection wavelength is 226nm;
Described condition of gradient elution is:
B. precision is drawn reference substance solution and need testing solution respectively, and injection liquid chromatography is measured pool horse degree amine and related substance thereof.
2. the method for simultaneously measuring pool horse degree amine and related substance thereof as claimed in claim 1, it is characterized in that: described step (1) preparation system applicability solution: get pool horse degree amine and accessory substance 5-amino-2-(2,6-dioxo-piperidines-3-yl)-isoindoline-1,3-diketone work reference substance, accurately weighed, adding acetonitrile makes to dissolve in right amount, add mobile phase A: it is all the solution of 0.01mg containing pool horse degree amine, accessory substance that the acetic acid that pH value is 2.6～3.0 is made every 1ml, the final volume of described acetonitrile and mobile phase A acetic acid is than being 20:80 again.
3. the method for simultaneously measuring pool horse degree amine and related substance thereof as claimed in claim 1, it is characterized in that: described step (2) preparation pool horse degree amine need testing solution: get pool horse degree amine, accurately weighed, adding acetonitrile makes to dissolve in right amount, add mobile phase A: the acetic acid that pH value is 2.6～3.0 is made the solution that every 1ml is 0.125mg containing pool horse degree amine, the final volume of described acetonitrile and mobile phase A acetic acid is than being 20:80.
4. in the time of as described in as arbitrary in claim 1～3, measure the method for pool horse degree amine and related substance thereof, it is characterized in that: described in step 3, the pH of acetum is 2.6.
5. in the time of as described in as arbitrary in claim 1～3, measure the method for pool horse degree amine and related substance thereof, it is characterized in that: described a. be take octadecylsilane chemically bonded silica as 150 * 4.0mm, 5 μ m filling agents, take acetum as mobile phase A, take first as Mobile phase B, gradient elution, column temperature is 30 ℃, flow velocity is 1ml/min, and detection wavelength is 226nm.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104749289A (en) * | 2015-04-16 | 2015-07-01 | 南京卡文迪许生物工程技术有限公司 | Method for detecting and separating pomalidomide related substances by use of high performance liquid chromatography |
| CN105334270A (en) * | 2014-06-30 | 2016-02-17 | 深圳海王药业有限公司 | Liquid chromatography for separating and determining pomalidomide intermediate related substance |
| CN105866297A (en) * | 2016-06-24 | 2016-08-17 | 合肥久诺医药科技有限公司 | High performance liquid chromatography method of pomalidomide related substance |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101535291A (en) * | 2006-09-26 | 2009-09-16 | 细胞基因公司 | 5-substituted quinazolinone derivatives used as antineoplastic drug |
| WO2013106686A1 (en) * | 2012-01-13 | 2013-07-18 | Celgene Corporation | Biomarkers for the treatment of hepatocellular carcinoma |
-
2013
- 2013-12-12 CN CN201310683222.3A patent/CN103645259B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101535291A (en) * | 2006-09-26 | 2009-09-16 | 细胞基因公司 | 5-substituted quinazolinone derivatives used as antineoplastic drug |
| WO2013106686A1 (en) * | 2012-01-13 | 2013-07-18 | Celgene Corporation | Biomarkers for the treatment of hepatocellular carcinoma |
Non-Patent Citations (3)
| Title |
|---|
| MATTHEW HOFFMANN 等: "Absorption, metabolism and excretion of [14C]pomalidomide in humans following oral administration", 《CANCER CHEMOTHER PHARMACOL》, vol. 71, no. 2, 28 February 2013 (2013-02-28), pages 489 - 501 * |
| PADIGELA SWETHA 等: "Estimation of pomalidomide in capsule dosage form by RP-HPLC", 《DER PHARMACIA LETTRE》, vol. 5, no. 3, 31 March 2013 (2013-03-31), pages 397 - 404 * |
| 李海望 等: "泊马度胺( Pomalidomide)", 《中国药物化学杂志》, vol. 23, no. 4, 31 August 2013 (2013-08-31), pages 339 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105334270A (en) * | 2014-06-30 | 2016-02-17 | 深圳海王药业有限公司 | Liquid chromatography for separating and determining pomalidomide intermediate related substance |
| CN104749289A (en) * | 2015-04-16 | 2015-07-01 | 南京卡文迪许生物工程技术有限公司 | Method for detecting and separating pomalidomide related substances by use of high performance liquid chromatography |
| CN105866297A (en) * | 2016-06-24 | 2016-08-17 | 合肥久诺医药科技有限公司 | High performance liquid chromatography method of pomalidomide related substance |
| CN105866297B (en) * | 2016-06-24 | 2019-01-04 | 合肥久诺医药科技有限公司 | A kind of HPLC analytical method of the pomalidomide in relation to substance |
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