CN106442751A - Method for determining content of conivaptan hydrochloride by virtue of high performance liquid chromatography - Google Patents
Method for determining content of conivaptan hydrochloride by virtue of high performance liquid chromatography Download PDFInfo
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- CN106442751A CN106442751A CN201610714667.7A CN201610714667A CN106442751A CN 106442751 A CN106442751 A CN 106442751A CN 201610714667 A CN201610714667 A CN 201610714667A CN 106442751 A CN106442751 A CN 106442751A
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- solution
- mobile phase
- conivaptan
- reference substance
- hydrochloric acid
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- 238000000034 method Methods 0.000 title claims abstract description 39
- 238000004128 high performance liquid chromatography Methods 0.000 title abstract description 10
- BTYHAFSDANBVMJ-UHFFFAOYSA-N conivaptan hydrochloride Chemical compound Cl.C12=CC=CC=C2C=2NC(C)=NC=2CCN1C(=O)C(C=C1)=CC=C1NC(=O)C1=CC=CC=C1C1=CC=CC=C1 BTYHAFSDANBVMJ-UHFFFAOYSA-N 0.000 title abstract description 6
- 229960004092 conivaptan hydrochloride Drugs 0.000 title abstract description 4
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims abstract description 27
- 229910000403 monosodium phosphate Inorganic materials 0.000 claims abstract description 13
- 235000019799 monosodium phosphate Nutrition 0.000 claims abstract description 13
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 claims abstract description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 103
- 239000000243 solution Substances 0.000 claims description 74
- JGBBVDFNZSRLIF-UHFFFAOYSA-N conivaptan Chemical compound C12=CC=CC=C2C=2[N]C(C)=NC=2CCN1C(=O)C(C=C1)=CC=C1NC(=O)C1=CC=CC=C1C1=CC=CC=C1 JGBBVDFNZSRLIF-UHFFFAOYSA-N 0.000 claims description 59
- 229960000562 conivaptan Drugs 0.000 claims description 59
- 238000012360 testing method Methods 0.000 claims description 46
- 239000013558 reference substance Substances 0.000 claims description 36
- 239000008363 phosphate buffer Substances 0.000 claims description 24
- 238000002360 preparation method Methods 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 14
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 claims description 9
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 8
- 238000001514 detection method Methods 0.000 claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 238000010521 absorption reaction Methods 0.000 claims description 6
- 238000010828 elution Methods 0.000 claims description 6
- 238000010829 isocratic elution Methods 0.000 claims description 6
- 238000004811 liquid chromatography Methods 0.000 claims description 6
- 238000005070 sampling Methods 0.000 claims description 5
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 4
- 238000010812 external standard method Methods 0.000 claims description 4
- BLRPTPMANUNPDV-UHFFFAOYSA-N Silane Chemical group [SiH4] BLRPTPMANUNPDV-UHFFFAOYSA-N 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- YTJSFYQNRXLOIC-UHFFFAOYSA-N octadecylsilane Chemical compound CCCCCCCCCCCCCCCCCC[SiH3] YTJSFYQNRXLOIC-UHFFFAOYSA-N 0.000 claims description 2
- 229910002027 silica gel Inorganic materials 0.000 claims description 2
- 239000000741 silica gel Substances 0.000 claims description 2
- 239000000377 silicon dioxide Substances 0.000 claims description 2
- 229960001866 silicon dioxide Drugs 0.000 claims description 2
- 230000003595 spectral effect Effects 0.000 claims description 2
- 238000007865 diluting Methods 0.000 claims 2
- 239000000872 buffer Substances 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 11
- 229940079593 drug Drugs 0.000 abstract description 10
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000008055 phosphate buffer solution Substances 0.000 abstract 4
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 abstract 1
- 230000035945 sensitivity Effects 0.000 abstract 1
- 235000019333 sodium laurylsulphate Nutrition 0.000 abstract 1
- 239000012071 phase Substances 0.000 description 27
- 239000002904 solvent Substances 0.000 description 20
- 239000000047 product Substances 0.000 description 14
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 9
- 238000003556 assay Methods 0.000 description 9
- 239000000203 mixture Substances 0.000 description 6
- VLTRZXGMWDSKGL-UHFFFAOYSA-N perchloric acid Chemical compound OCl(=O)(=O)=O VLTRZXGMWDSKGL-UHFFFAOYSA-N 0.000 description 6
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 238000003918 potentiometric titration Methods 0.000 description 4
- RZYKUPXRYIOEME-UHFFFAOYSA-N CCCCCCCCCCCC[S] Chemical compound CCCCCCCCCCCC[S] RZYKUPXRYIOEME-UHFFFAOYSA-N 0.000 description 3
- 206010021036 Hyponatraemia Diseases 0.000 description 3
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 3
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000011574 phosphorus Substances 0.000 description 3
- 229910052698 phosphorus Inorganic materials 0.000 description 3
- 239000011734 sodium Substances 0.000 description 3
- 229910052708 sodium Inorganic materials 0.000 description 3
- 238000004448 titration Methods 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
- 229960000583 acetic acid Drugs 0.000 description 2
- FGDQGIKMWOAFIK-UHFFFAOYSA-N acetonitrile;phosphoric acid Chemical group CC#N.OP(O)(O)=O FGDQGIKMWOAFIK-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000012417 linear regression Methods 0.000 description 2
- 239000007791 liquid phase Substances 0.000 description 2
- BRMYZIKAHFEUFJ-UHFFFAOYSA-L mercury diacetate Chemical compound CC(=O)O[Hg]OC(C)=O BRMYZIKAHFEUFJ-UHFFFAOYSA-L 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012430 stability testing Methods 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- VWSLLSXLURJCDF-UHFFFAOYSA-N 2-methyl-4,5-dihydro-1h-imidazole Chemical compound CC1=NCCN1 VWSLLSXLURJCDF-UHFFFAOYSA-N 0.000 description 1
- 206010020919 Hypervolaemia Diseases 0.000 description 1
- 208000019693 Lung disease Diseases 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- KBZOIRJILGZLEJ-LGYYRGKSSA-N argipressin Chemical compound C([C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CSSC[C@@H](C(N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N1)=O)N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(N)=O)C1=CC=CC=C1 KBZOIRJILGZLEJ-LGYYRGKSSA-N 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 238000011088 calibration curve Methods 0.000 description 1
- 125000002915 carbonyl group Chemical group [*:2]C([*:1])=O 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 208000003532 hypothyroidism Diseases 0.000 description 1
- 230000002989 hypothyroidism Effects 0.000 description 1
- 201000008284 inappropriate ADH syndrome Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000003182 parenteral nutrition solution Substances 0.000 description 1
- 238000005220 pharmaceutical analysis Methods 0.000 description 1
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000005063 solubilization Methods 0.000 description 1
- 230000007928 solubilization Effects 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229940054353 vaprisol Drugs 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- 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
- G01N2030/022—Column chromatography characterised by the kind of separation mechanism
- G01N2030/027—Liquid chromatography
Landscapes
- Physics & Mathematics (AREA)
- Spectroscopy & Molecular Physics (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)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention provides a method for determining the content of conivaptan hydrochloride by virtue of a high performance liquid chromatography. The method comprises the following chromatographic conditions: a mobile phase A is acetonitrile, a mobile phase B is a phosphate buffer solution, the concentration of sodium dihydrogen phosphate in the phosphate buffer solution is 0.02mol/L, the mass concentration of sodium lauryl sulfate in the phosphate buffer solution is 0.5%, and the pH value of the phosphate buffer solution is 2.8-3.2. By virtue of the method, the content of a conivaptan hydrochloride raw material drug can be effectively determined, and the method is high in accuracy and sensitivity and good in repeatability. Compared with a traditional method, the method has the characteristics of high efficiency and accuracy.
Description
Technical field
The present invention relates to Pharmaceutical Analysis technical field is and in particular to one kind examines Buddhist nun using high effective liquid chromatography for measuring hydrochloric acid
The method cutting down smooth bulk drug content.
Background technology
The entitled N- of chemistry { 4- [(lysidine simultaneously [4,5-d] [the 1]-benzo-aza of hydrochloric acid conivaptan
Tall and erect -6 (1H)-yls) carbonyl] phenyl } xenyl -2- carboxamide hydrochloride, No. CAS is 168626-94-6, and English name is
Conivaptan Hydrochloride.Hydrochloric acid conivaptan is that one kind of arginine vasopressin (AVP) V1a and V2 acceptor is non-peptide
Class double inhibitor.FDA ratified the hydrochloric acid conivaptan of Astellas Pharma US company early than on December 29th, 2005
Listing, trade name Vaprisol, is the parenteral solution of 20mg/4mL, and its indication is mainly used in the normal hyponatremia of blood volume
(often occur together in syndrome of inappropriate secretion of antidiuretic hormone patient, patients with hypothyroidism, hyposupraenalism patient or
Pulmonary Disease patients) and high power capacity hyponatremia inpatient treatment.FDA approval conivaptan in 2007 increases and adapts to
Disease Hypervolemia hyponatremia, ratifies to increase the big transfusion of new formulation -20mg/100mL for 2008 again.
Prior art adopts potentiometric determination hydrochloric acid conivaptan bulk drug content.Formic acid, ice in the method
The reagent such as acetic acid, mercuric acetate, perchloric acid, toxicity is relatively large, in addition operates also loaded down with trivial details, particularly many batches of raw materials is examined simultaneously
Survey, the amount of labour is also to be doubled and redoubled.
For this reason, it may be necessary to find a kind of effectively reduction workload, agents useful for same small toxicity or consumption are few, simultaneously can be accurate
The method really detecting its content.
Content of the invention
For overcoming prior art not enough, the present invention provides a kind of former using high effective liquid chromatography for measuring hydrochloric acid conivaptan
The method of material medicine content, in order to control product quality.
Technical solution of the present invention is as follows：
A kind of method of employing high effective liquid chromatography for measuring hydrochloric acid conivaptan content, including following chromatographic condition：
Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer；Wherein, in described phosphate buffer sodium dihydrogen phosphate dense
Spending the mass concentration for 0.02mol/L, lauryl sodium sulfate is 0.5%, pH value 2.8-3.2.
Preferably, mobile phase A and the volume ratio of Mobile phase B are 55-59:45-41, such as 55:45、59:41 or 57:43,
It is still more preferably 57:43.
Preferably, described phosphate buffer pH value 3.0.
The preparation method of described phosphate buffer includes appropriate sodium dihydrogen phosphate and lauryl sodium sulfate are dissolved in water,
Adjusted to required pH value with phosphoric acid.
Preferably, chromatographic column used is silane group silicagel column, more preferably octadecylsilane chemically bonded silica post
(C18 chromatographic column)；It is still more preferably
C18,5 μm of 250 × 4.6mm (Cat.no. of specification:99603) or
Phenomenex C18,5 μm of 250 × 4.6mm of specification or
Agilent C18,5 μm of 250 × 4.6mm of specification.
Preferably, type of elution is isocratic elution.
Preferably, flow rate of mobile phase 0.9-1.1mL/min, further preferred 1.0mL/min.
Preferably, 22-28 DEG C of column temperature, further preferred 25 DEG C；
Preferably, sampling volume：10μL；
Detector is UV-detector；It is preferably Agilent G1315B DAD detector or Agilent G1315B VWD
Detector.
Preferably, Detection wavelength is 200-280nm, more preferably 204nm ± 2nm, 241nm ± 2nm or 276nm ±
2nm；It is still more preferably 240nm.
Said determination method, wherein：
Preferably, it is diluted to need testing solution with mobile phase respectively including by hydrochloric acid conivaptan testing sample and reference substance
And reference substance solution, then detected.
Preferably, using Agilent 1260 high performance liquid chromatograph or Agilent 1100 high performance liquid chromatograph.
Preferably, using Agilent Chemstation chromatographic work station.
Said determination method, also includes the preparation of need testing solution, the preparation of reference substance solution.
The preparation of described need testing solution includes：Take sample appropriate, accurately weighed, solubilizer dissolve and dilute make containing about
The solution of hydrochloric acid conivaptan 0.2mg/mL, as need testing solution.
The preparation of described reference substance solution includes：Take hydrochloric acid conivaptan reference substance appropriate, accurately weighed, solubilizer dissolves
And dilute the solution making about hydrochloric conivaptan 0.2mg/mL, as reference substance solution.
The above-mentioned need testing solution and the solvent of reference substance solution prepared is above-mentioned mobile phase, specially by mobile phase A and stream
According to the initial volume eluting, than the mixture forming, (mobile phase A is 55-59 with the volume ratio of B to dynamic phase B:45-41, such as 55:
45、59:41 or 57:43, preferably 57:43).
The method that the present invention adopts high effective liquid chromatography for measuring hydrochloric acid conivaptan content, specifically includes following steps：
1) following chromatographic condition is set：
Chromatographic column：C18,5 μm of 250 × 4.6mm (Cat.no. of specification:99603)；Or
Phenomenex C18,5 μm of 250 × 4.6mm of specification；Or Agilent C18,5 μm of 250 × 4.6mm of specification；
Mobile phase：Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer, and the two volume ratio is 57:43；
Wherein, the preparation method of described phosphate buffer include sodium dihydrogen phosphate and lauryl sodium sulfate add water molten
Solution, with phosphorus acid for adjusting pH value to 3.0；In described phosphate buffer, the concentration of sodium dihydrogen phosphate is 0.02mol/L, dodecyl sulphur
The mass concentration of sour sodium is 0.5%；
Type of elution is isocratic elution；
Flow rate of mobile phase：1.0mL/min, column temperature：25 DEG C, sampling volume：10μL；
Detector is Agilent G1315B DAD detector or Agilent G1315B VWD detector；
Detection wavelength is 240nm；
2) preparation of need testing solution：Take sample appropriate, accurately weighed, solubilizer dissolves and dilutes makes about hydrochloric examining
Buddhist nun cuts down the solution of smooth 0.2mg/mL, as need testing solution；
The preparation of reference substance solution：Take hydrochloric acid conivaptan reference substance appropriate, accurately weighed, solubilizer dissolves and dilutes system
Become the solution of about hydrochloric conivaptan 0.2mg/mL, as reference substance solution；
Described solvent is according to volume ratio 57 by mobile phase A with Mobile phase B:The mixture of 43 compositions；
3) detect：Accurate absorption reference substance solution and each 10 μ L of need testing solution, inject high performance liquid chromatograph, record color
Spectrogram, by external standard method with the content of hydrochloric acid conivaptan in calculated by peak area test sample.
The invention has the advantages that：
High-efficient liquid phase chromatogram condition is optimized the present invention it is determined that optimal detection condition.According to product performance,
Prioritizing selection acid condition in the selection of mobile phase, allows hydrochloric acid conivaptan to exist with molecular conformation as far as possible it is ensured that ultraviolet is inhaled
The completeness received.Especially in accordance with product performance, find out optimal organic phase-inorganic matched.
The method that the present invention adopts can effectively measure the content of hydrochloric acid conivaptan bulk drug, the method degree of accuracy and sensitive
Degree is high, favorable reproducibility.The method is applied to the assay of hydrochloric acid conivaptan bulk drug, solves current hydrochloric acid conivaptan
The unstable problem of content is it is adaptable to the quality control of hydrochloric acid conivaptan product.
Brief description
Fig. 1 is the HPLC chromatogram of solvent in experimental example 2；
Fig. 2 is the HPLC chromatogram of hydrochloric acid conivaptan reference substance in experimental example 2；
Fig. 3 is the HPLC chromatogram of hydrochloric acid conivaptan test sample in experimental example 2；
Fig. 4 is experimental example 3 hydrochloric acid conivaptan canonical plotting.
Specific embodiment
Following examples are used for the present invention is described, but are not limited to the scope of the present invention.Unreceipted concrete in embodiment
Technology or condition person, according to the technology described by document in the art or condition, or are carried out according to product description.Used
Reagent or the unreceipted production firm person of instrument, are the conventional products being commercially available by regular distributor.
Wherein, hydrochloric acid conivaptan bulk drug be purchased from Anqing Ward medication chemistry Co., Ltd (containing lot number 20130129,
20130323、20130518、20130520、20130524).Described hydrochloric acid conivaptan reference substance is purchased from Shenzhen Fes biology section
Skill Co., Ltd, lot number 1312326-94-6
If no special instructions, concentration is generally referred to as the w/v g/mL of solute and solution to the present invention.If solute
It is liquid, then concentration is volume ratio, such as 0.1% trifluoroacetic acid aqueous solution refers to that the trifluoroacetic acid of 0.1mL is dissolved in the water of 100mL
In.
Described chemical reagent purity is chromatographically pure rank, purchased from Guangzhou Xin Hong trade Co., Ltd (imported from America).
Described high performance liquid chromatograph is：Agilent 1260 high performance liquid chromatograph, Agilent 1100 efficient liquid phase
Chromatograph.
Detector：Agilent G1315B DAD detector Agilent G1315B VWD detector.
Chromatographic work station：Agilent Chemstation chromatographic work station.
If no special indicate, described below solvent is acetonitrile-phosphate buffer, and acetonitrile with the volume ratio of phosphate buffer is
57:43.The preparation method of described phosphate buffer includes sodium dihydrogen phosphate and lauryl sodium sulfate are dissolved in water, and uses phosphoric acid
Adjust pH value to 3.0；In described phosphate buffer, the concentration of sodium dihydrogen phosphate is 0.02mol/L, the matter of lauryl sodium sulfate
Amount concentration is 0.5%.
If no special indicate, using described below chromatographic condition：
Chromatographic column：C18,5 μm of 250 × 4.6mm (Cat.no. of specification:99603) or Phenomenex
C18, specification 5 μm of 250 × 4.6mm or Agilent C18,5 μm of 250 × 4.6mm of specification；
Mobile phase：Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer, and the two volume ratio is 57:43；
Wherein, the preparation method of described phosphate buffer include sodium dihydrogen phosphate and lauryl sodium sulfate add water molten
Solution, with phosphorus acid for adjusting pH value to 3.0；In described phosphate buffer, the concentration of sodium dihydrogen phosphate is 0.02mol/L, dodecyl sulphur
The mass concentration of sour sodium is 0.5%；
Type of elution is isocratic elution；
Flow velocity：1.0mL/min, column temperature：25 DEG C, sampling volume：10μL；
Detector is Agilent G1315B DAD detector or Agilent G1315B VWD detector；
Detection wavelength is 240nm.
Experimental example 1 maximum absorption wavelength and the selection of chromatographic condition
Take hydrochloric acid conivaptan bulk drug, plus methyl alcohol makes in every 1 ml methanol the about hydrochloric acid conivaptan containing 20 μ g
Solution, according to ultraviolet spectrophotometry (2010 editions two annex IV A of Chinese Pharmacopoeia) measure, in 200-400nm wave-length coverage
Carry out length scanning, result of the test see table 1.
Table 1 length scanning result
Result of the test shows that the methanol solution of this product has at 204nm ± 2nm, 241nm ± 2nm, 276nm ± 2nm wavelength
Absorption maximum, sample is consistent with the ultraviolet absorpting spectrum of reference substance.It is easier to be interfered in view of 204nm, 276nm absworption peak is not
Substantially, thus preferably employ 240nm as the assay absorbing wavelength of this product.
Experimental example 2 specificity
A, reference substance solution：Take hydrochloric acid conivaptan reference substance (lot number：20130223) about 20mg, accurately weighed, puts
In 100mL measuring bottle, (i.e. acetonitrile-phosphate buffer, acetonitrile is 57 with the volume ratio of phosphate buffer to solubilizer:43) dissolving and dilute
Release to scale, shake up, obtain final product.
B, need testing solution：Take this product (lot number：20130129) about 20mg, accurately weighed, puts in 100mL measuring bottle, solubilization
Agent is dissolved and is diluted to scale, shakes up, and obtains final product.
Take each 10 μ L injection high performance liquid chromatographs of solvent, reference substance solution, need testing solution, record chromatogram is (respectively
See Fig. 1-3)；Result display need testing solution and reference substance solution are in identical position appearance, and solvent does not produce in this position
Any chromatographic peak, illustrates that solvent does not disturb the mensure of hydrochloric acid conivaptan in this product.
Experimental example 3 is linear and scope
Take the hydrochloric acid conivaptan reference substance 50.12mg being dried to constant weight, accurately weighed, put in 25mL measuring bottle, use solvent
Dissolved dilution, to scale, shakes up, as reference substance storing solution；Precision measure this reference substance storing solution 0.4mL, 0.8mL, 1.0mL,
1.2mL, 1.4mL and 1.6mL put in 10mL measuring bottle respectively, and solubilizer is diluted to scale, shake up, under above-mentioned chromatographic condition, essence
Close measure each solution 10 μ L injection high performance liquid chromatograph, record chromatogram；With sample introduction concentration (mg/mL) as X, hydrochloric acid is examined Buddhist nun and is cut down
Smooth peak area (A) is Y, makees equation of linear regression, result such as table 2 below.Fig. 4 is hydrochloric acid conivaptan calibration curve.
The preparation of table 2 hydrochloric acid conivaptan content measuring standard curve
Conclusion：Hydrochloric acid conivaptan sample size with peak area is in the range of 0.08019mg/mL～0.320768mg/mL
Good linear relationship, equation of linear regression is：Y=36100x+92.11, y represent peak area, and x represents sample introduction concentration (mg/
mL)；R2=0.999.
Experimental example 4 precision test
A, sample introduction precision test
The accurate reference substance solution 10 μ L injection high performance liquid chromatograph drawing experimental example 3 intermediate concentration, continuous sample introduction 7
Pin, records chromatogram, investigates the situation of change of hydrochloric acid conivaptan main peak peak area, and result see table 3.
Table 3 hydrochloric acid conivaptan sample introduction Precision test result (n=6)
Conclusion：Sample introduction precision meets regulation.
B, replica test
Take same batch sample (lot number：20130129), accurately weighed appropriate, solubilizer dissolves and dilutes to be made in every 1mL
The solution of about hydrochloric conivaptan 0.2mg, as need testing solution, 6 parts of parallel preparation.Separately take hydrochloric acid conivaptan reference substance
In right amount, accurately weighed, solubilizer dissolves and dilutes the solution making about hydrochloric conivaptan 0.2mg in every 1mL, as comparison
The each 10 μ L injection liquid chromatographs of product solution, accurate absorption reference substance solution and need testing solution, record chromatogram；By external standard method
With calculated by peak area, result such as table 4 below.
Table 4 hydrochloric acid conivaptan replica test result
Conclusion：This experimental technique repeatability meets regulation.
C, Intermediate precision test
Prepare solution according under " b, replica test " item, by two analysis different instruments of librarian use, when different
Between tested, gained assay data, result such as table 5 below：
Table 5 hydrochloric acid conivaptan Intermediate precision result of the test
Conclusion：This method Intermediate precision is good.
Experimental example 5 assay solution stability testing to be measured
Take hydrochloric acid conivaptan bulk drug appropriate, accurately weighed, solubilizer dissolve and dilute make about hydrochloric in every 1mL
The solution of conivaptan 0.2mg, as need testing solution.Measure in 0,2,4,6,8,10,12 and 24 hours sample introductions respectively, investigate
The situation of change of need testing solution main peak area, result of the test see table 6：
Table 6 hydrochloric acid conivaptan assay solution stability testing to be measured
Conclusion：From above-mentioned result of the test, need testing solution was stable in 24 hours.
Experimental example 6 serviceability test
Parameter in chromatographic condition such as flow velocity, column temperature, flowing phase pH value, mobile phase ratio and different chromatographic columns etc. are made
Suitable change, investigates mass-tone spectral peak situation in hydrochloric acid conivaptan, and result see table 7.
Table 7 serviceability test
Conclusion：After parameter in chromatographic condition is made suitably to change, the peak area of hydrochloric acid conivaptan main peak does not produce
Significantly change, this chromatographic condition has preferable durability.
Comparative example 1
It is respectively adopted HPLC method of the present invention and potentiometric titration, hydrochloric acid conivaptan bulk drug content is carried out with contrast inspection
Test is tested.Understand the otherness of two methods.The results are shown in Table down 8.
Table 8 two methods measurement result compares
HPLC method of the present invention comprises the following steps：
1) following chromatographic condition is set：
Chromatographic column：C18,5 μm of 250 × 4.6mm (Cat.no. of specification:99603) or Phenomenex
C18, specification 5 μm of 250 × 4.6mm or Agilent C18,5 μm of 250 × 4.6mm of specification；
Mobile phase：Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer, and the two volume ratio is 57:43；
Wherein, the preparation method of described phosphate buffer include sodium dihydrogen phosphate and lauryl sodium sulfate add water molten
Solution, with phosphorus acid for adjusting pH value to 3.0；In described phosphate buffer, the concentration of sodium dihydrogen phosphate is 0.02mol/L, dodecyl sulphur
The mass concentration of sour sodium is 0.5%；
Type of elution is isocratic elution；Flow velocity：1.0mL/min, column temperature：25 DEG C, sampling volume：10μL；
Detector is Agilent G1315B DAD detector or Agilent G1315B VWD detector；Detection wavelength is
240nm；
2) preparation of need testing solution：Take sample appropriate, accurately weighed, solubilizer dissolves and dilutes makes about hydrochloric examining
Buddhist nun cuts down the solution of smooth 0.2mg/mL, as need testing solution；
The preparation of reference substance solution：Take hydrochloric acid conivaptan reference substance appropriate, accurately weighed, solubilizer dissolves and dilutes system
Become the solution of about hydrochloric conivaptan 0.2mg/mL, as reference substance solution；
Described solvent is according to volume ratio 57 by mobile phase A with Mobile phase B:The mixture of 43 compositions；
3) detect：Accurate absorption reference substance solution and each 10 μ L of need testing solution, inject high performance liquid chromatograph, record color
Spectrogram, by external standard method with the content of hydrochloric acid conivaptan in calculated by peak area test sample.
Potentiometric titration：Take this product about 0.4g, accurately weighed, plus after the dissolving of 5mL formic acid, add 20mL glacial acetic acid and 5mL
Mercuric acetate, uses 0.1mol/L perchloric acid titration drop fixed according to potentiometric titration (Chinese Pharmacopoeia two annex VII A of version in 2010), and
The result blank test of titration is corrected.Every 1mL perchloric acid titration liquid (0.1mol/L) is equivalent to hydrochloric acid conivaptan
(C32H26N4O2·HCl)53.504mg.
Table 8 result shows, this product HPLC method assay result is basically identical with potentiometric titration assay result, says
Bright two methods are all applied to the assay of hydrochloric acid conivaptan.Because of assay (HPLC method) and Related substances separation chromatogram
Consistent, for ease of operation, determine the content assaying method as this product using HPLC method.
Although, above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
Claims (10)
1. a kind of method of employing high effective liquid chromatography for measuring hydrochloric acid conivaptan content is it is characterised in that include following color
Spectral condition：Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer；Wherein, in described phosphate buffer sodium dihydrogen phosphate dense
Spending the mass concentration for 0.02mol/L, lauryl sodium sulfate is 0.5%, pH value 2.8-3.2.
2. method according to claim 1 is it is characterised in that described mobile phase A is 55-59 with the volume ratio of Mobile phase B:
45-41, preferably 55:45、59:41 or 57:43.
3. method according to claim 1 and 2 is it is characterised in that the preparation method of described phosphate buffer includes fitting
Amount sodium dihydrogen phosphate and lauryl sodium sulfate are dissolved in water, and are adjusted to required pH value with phosphoric acid；Preferably, described phosphoric acid buffer
The pH value of liquid is 3.0.
4. method according to claim 1 and 2 is it is characterised in that chromatographic column used is silane group silicagel column, preferably
Octadecylsilane chemically bonded silica post；More preferably
C18,5 μm of 250 × 4.6mm (Cat.no. of specification:99603)；Or,
Phenomenex C18,5 μm of 250 × 4.6mm of specification；Or,
Agilent C18,5 μm of 250 × 4.6mm of specification.
5. method according to claim 1 and 2 is it is characterised in that type of elution is isocratic elution；
And/or, flow rate of mobile phase 0.9-1.1mL/min, preferably 1.0mL/min.
6. method according to claim 1 and 2 is it is characterised in that 22-28 DEG C of column temperature, preferably 25 DEG C.
7. method according to claim 1 and 2 is it is characterised in that detector is UV-detector；It is preferably Agilent
G1315B DAD detector or Agilent G1315B VWD detector.
8. method according to claim 1 and 2 is it is characterised in that Detection wavelength is 200-280nm, preferably 240nm.
9. method according to claim 1 and 2 is it is characterised in that also include the preparation of need testing solution, reference substance solution
Preparation；
The preparation of described need testing solution includes：Take sample appropriate, accurately weighed, plus flowing phased soln dilute and make about saliferous
The solution of sour conivaptan 0.2mg/mL, as need testing solution；
The preparation of described reference substance solution includes：Take hydrochloric acid conivaptan reference substance appropriate, accurately weighed, plus flowing phased soln is simultaneously
The solution of about hydrochloric conivaptan 0.2mg/mL is made in dilution, as reference substance solution.
10. method according to claim 1 and 2 is it is characterised in that comprise the following steps：
1) following chromatographic condition is set：
Chromatographic column：C18,5 μm of 250 × 4.6mm of specification；Or, Phenomenex C18,5 μm 250 of specification ×
4.6mm；Or, Agilent C18,5 μm of 250 × 4.6mm of specification；
Mobile phase：Mobile phase A is acetonitrile, and Mobile phase B is phosphate buffer, and the two volume ratio is 57:43；Described phosphate buffer
PH value be 3.0；
Type of elution is isocratic elution；
Flow rate of mobile phase：1.0mL/min, column temperature：25 DEG C, sampling volume：10μL；
Detector is Agilent G1315B DAD detector or Agilent G1315B VWD detector；
Detection wavelength is 240nm；
2) preparation of need testing solution：Take sample appropriate, accurately weighed, plus flowing phased soln diluting makes and about hydrochloric examines Buddhist nun
Cut down the solution of smooth 0.2mg/mL, as need testing solution；
The preparation of reference substance solution：Take hydrochloric acid conivaptan reference substance appropriate, accurately weighed, plus flowing phased soln diluting makes
The solution of about hydrochloric conivaptan 0.2mg/mL, as reference substance solution；
3) detect：Accurate absorption reference substance solution and each 10 μ L of need testing solution, inject high performance liquid chromatograph, record chromatogram
Figure, by external standard method with the content of hydrochloric acid conivaptan in calculated by peak area test sample.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103497195A (en) * | 2013-10-21 | 2014-01-08 | 北京科莱博医药开发有限责任公司 | Conivaptan-hydrochloride novel crystal form and preparation method thereof |
| CN105310978A (en) * | 2014-08-04 | 2016-02-10 | 李峰 | Drug combination containing conivaptan hydrochloride as active ingredient and preparation of drug combination |
-
2016
- 2016-08-24 CN CN201610714667.7A patent/CN106442751A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103497195A (en) * | 2013-10-21 | 2014-01-08 | 北京科莱博医药开发有限责任公司 | Conivaptan-hydrochloride novel crystal form and preparation method thereof |
| CN105310978A (en) * | 2014-08-04 | 2016-02-10 | 李峰 | Drug combination containing conivaptan hydrochloride as active ingredient and preparation of drug combination |
Non-Patent Citations (3)
| Title |
|---|
| 蒋大圆 等: "HPLC法和非水电位滴定法测定盐酸考尼伐坦的含量", 《宜春学院学报》 * |
| 郑登宇 等: "盐酸考尼伐坦的合成", 《中国医药工业杂志》 * |
| 马亚娟 等: "盐酸考尼伐坦的研究进展", 《广东化工》 * |
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