CN113390999A - Control and detection method for sodium nitroprusside degradation impurities - Google Patents
Control and detection method for sodium nitroprusside degradation impurities Download PDFInfo
- Publication number
- CN113390999A CN113390999A CN202110707465.0A CN202110707465A CN113390999A CN 113390999 A CN113390999 A CN 113390999A CN 202110707465 A CN202110707465 A CN 202110707465A CN 113390999 A CN113390999 A CN 113390999A
- Authority
- CN
- China
- Prior art keywords
- sodium nitroprusside
- impurities
- sample
- liquid chromatography
- solution
- 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.)
- Pending
Links
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
- G01N30/50—Conditioning of the sorbent material or stationary liquid
- G01N30/52—Physical parameters
- G01N30/54—Temperature
-
- 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
Landscapes
- Physics & Mathematics (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Investigating Or Analyzing Non-Biological Materials By The Use Of Chemical Means (AREA)
Abstract
The invention provides an evaluation method of sodium nitroprusside stability, and discloses an analysis method for rapidly determining sodium nitroprusside degradation impurities. The method is characterized in that the method adopts high performance liquid chromatography for determination, adopts a chromatographic column with phenyl bonded silica gel as a filling agent, and takes a mixed solution of an ion pair reagent and an organic reagent as a mobile phase. The method has the advantages of strong specificity, good repeatability and simple and convenient operation, and can ensure the effective monitoring of the sodium nitroprusside and the degradation impurities in the preparation placing process.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry, particularly relates to the technical field of determination of related substances of medicines, and particularly relates to a control and detection method of sodium nitroprusside degradation impurities.
Background
Sodium nitroprusside, a nitrohydrocyanate.
Sodium nitroprusside is a vasodilator and is a common medicine for treating hypertensive emergency and acute left heart failure. The composition has direct dilating effect on resistance and volume blood vessel, and has effect on afterload greater than nitroglycerin, so as to reduce left ventricular filling pressure and increase cardiac output. For acute decompensation of chronic left ventricular failure patients, the sodium nitroprusside has quick and strong effect. The action mechanism is the same as that of nitrate, and the nitrate can enable vascular endothelial cells to release NO and activate guanylate cyclase, increase the intracellular cGMP level and dilate blood vessels. During metabolism, it is first converted into cyanide by erythrocytes and then converted into the end metabolite thiocyanate by the enzyme rhodanese in the liver. The thiocyanic acid is discharged from the kidney, and the half-life period of the patient with normal renal function is 4-7 days.
The sodium nitroprusside is convenient to use, takes effect quickly after intravenous drip, has quick disappearance, generally does not need the monitoring of traumatic hemodynamics, and is a preferred medicament for various hypertension emergencies.
The half-life of sodium nitroprusside is extremely short. The action is maintained for only 5-15 min, so the curative effect should be maintained by intravenous drip.
Sodium nitroprusside needs to be freshly prepared for intravenous administration. When the sodium nitroprusside is prepared with sodium chloride injection and 5 percent glucose injection, the stability of the sodium nitroprusside is reduced rapidly along with the prolonging of time.
Sodium nitroprusside is unstable in a solution state, and is usually observed through the change of items such as content, appearance color, pH value, visible foreign matters, insoluble particles and the like, and the change trend of degraded impurities is rarely concerned.
The control of related substances of sodium nitroprusside and related preparations is also shown to be extremely important.
The sodium nitroprusside has weaker ultraviolet response, and the degradation impurities of the sodium nitroprusside are mainly molecular ionic compounds with large polarity. The above features present challenges to conventional detection methods.
In the research, the liquid chromatography is adopted for control, the operation is simple, and the main degradation impurities of the sodium nitroprusside can be effectively detected.
Disclosure of Invention
The invention aims to provide a method for separating and measuring trace impurities in sodium nitroprusside by using a liquid chromatography, and an ultraviolet detector is adopted for measuring. The method can effectively achieve the separation and determination of various degradation impurities, thereby realizing the effective control of the quality of the sodium nitroprusside. Solves the problem of quality control of sodium nitroprusside degradation impurities in the prior art, and the method is simple, rapid and accurate. The method is characterized in that:
the mobile phase is phosphate buffer solution, acetonitrile or methanol, preferably 0.01-0.1 mol/L phosphate solution and acetonitrile, and more preferably 0.02mol/L phosphate buffer solution.
The ion-pairing agent in the mobile phase may be trifluoroacetic acid, tetrabutylammonium hydroxide, tetrabutylammonium bromide and tetrabutylammonium hydrogen sulfate, preferably tetrabutylammonium hydrogen sulfate.
The water phase in the mobile phase is phosphate buffer solution which contains 0.1-1% of ion pair reagent, and the pH value is adjusted to 3.0-6.0 by phosphoric acid. The method preferably comprises 0.2-0.4% of ion-pairing agent, and the pH value is 5.0-6.0.
The diluent is the initial mobile phase or water, preferably water.
The concentration of the sodium nitroprusside in the test sample is 5-25 mg/ml, preferably 10-20 mg/ml.
The method adopts chromatographic columns with octadecylsilane chemically bonded silica and phenyl chemically bonded silica as fillers, and preferably adopts chromatographic columns with phenyl chemically bonded silica as fillers.
The method is characterized in that the flow velocity of the mobile phase is set to be 1.0-2.0 ml/min, and the column temperature is controlled to be 35-50 ℃.
In the method, an ultraviolet detector is adopted, and the detection wavelength is preferably 210-230 nm.
The specific method for controlling and measuring the degradation impurities in the sodium nitroprusside is as follows:
instrument and reagent
Shijin 2030plus high performance liquid chromatograph, Xiang apparatus L420 type centrifuge, permanent lighting box, acetonitrile (pure chromatogram), hydrochloric acid (pure chromatogram), sodium hydroxide (pure chromatogram), tetrabutyl ammonium hydrogen sulfate (pure chromatogram).
Diluent agent: and (3) water.
Mobile phase, phosphate buffer: dissolving 3g tetrabutylammonium hydrogen sulfate and 2.8g anhydrous disodium hydrogen phosphate in 1000ml water, and adjusting the pH value to 6.0 by phosphoric acid;
control solution: taking a proper amount of sodium nitroprusside, a nitrous acid solution, a nitric acid solution, a ferrocyanide compound and a ferricyanide compound as reference substances, quantitatively diluting with water, and shaking up to obtain a corresponding reference substance solution.
Secondly, measuring the sample:
(1) pretreatment of a test sample: performing operation in a dark place, taking a sodium nitroprusside sample, precisely weighing, and adding water to dissolve the sodium nitroprusside sample to obtain a test solution; performing operation in a dark place, taking a sodium nitroprusside sample, precisely weighing, adding water to dissolve, and placing in a water bath at 60 ℃ for 4 hours to serve as a high-temperature damage test sample solution; and (3) operating in a dark place, taking a sodium nitroprusside sample, precisely weighing, adding water to dissolve, and placing the sodium nitroprusside sample in a 5000lx light condition for 2 hours to serve as a strong light damage test sample solution.
(2) Chromatographic conditions are as follows:
mobile phase: the phosphate buffer solution is used as a water phase, the methanol is used as an organic phase, and gradient elution is carried out.
The ultraviolet detection wavelength is 230nm by adopting high performance liquid chromatography for determination.
(3) Specificity test
Under the chromatographic conditions, taking blank solvent, impurity reference substances and test solution with strong sodium nitroprusside destruction in turn, and recording chromatogram. According to the detection result, the separation degrees between the sodium nitroprusside and each known impurity meet the requirement, the separation degrees between the sodium nitroprusside and the adjacent chromatographic peak under the strong degradation condition are both larger than 1.5, the blank solvent interferes the detection, and the specificity meets the requirement.
TABLE 1 Strong degradation test results
(4) Limit of quantification
Under the chromatographic conditions described above, the limit of quantitation was determined from the response concentration at which the signal-to-noise ratio of the chromatographic peak was about 10: 1. The limit of quantitation of the ferricyanide compound was determined to be about 0.3 μ g/ml; the limit of quantitation of the ferrocyanide compound is about 0.05 μ g/ml; the quantitative limit concentration of nitrous acid impurities is about 0.05 mu g/ml; the limiting concentration of nitric acid impurity was about 0.25. mu.g/ml.
(5) Linearity
Preparing reference substance solutions with different concentrations, respectively carrying out sample injection detection, recording peak areas, carrying out linear regression by taking the concentration as a horizontal coordinate and the peak area as a vertical coordinate to obtain a regression equation, wherein the result is shown in table 2.
TABLE 2 Linear measurement results
(6) Precision test of instrument
One part of sodium nitroprusside reference substance solution with quantitative limit concentration is prepared, 6 needles are continuously injected according to the conditions, the peak area is recorded, the standard average deviation of the relative peak area is calculated to be 1.8 percent, and the result is shown in table 3.
TABLE 3 measurement of precision of sodium nitroprusside
| |
1 | 2 | 3 | 4 | 5 | 6 | RSD |
| Peak area | 11235 | 11178 | 11105 | 11104 | 10887 | 10712 | 1.8％ |
(7) Stability of the solution
Taking sodium nitroprusside test solution and self-control solution, placing the sodium nitroprusside test solution and the self-control solution under the conditions of light shielding and no light shielding, respectively injecting and measuring, and inspecting the change of the solutions, wherein the results are shown in table 4. Sodium nitroprusside is sensitive to light in an aqueous solution state, and impurities are increased quickly; under the condition of keeping out of the light, the impurities are not obviously increased.
TABLE 4 stability assay results for sodium nitroprusside solution
The method has the characteristics of strong specificity, high sensitivity, wide linear range, good solution stability under the condition of light shielding and the like. Provides powerful scientific basis for feasible determination of degraded impurities in the sodium nitroprusside raw material and the preparation.
The invention has the following positive effects: the invention provides a high performance liquid chromatography for monitoring various degradation impurities of sodium nitroprusside, which provides a reference for optimizing detection methods of sodium nitroprusside and preparations thereof. The invention adopts high performance liquid chromatography for determination, and particularly obtains special chromatographic conditions through creative labor, thereby realizing the rapid monitoring of the change trend of impurities in sodium nitroprusside, and finally ensuring the effectiveness and safety of the sodium nitroprusside.
Drawings
FIG. 1 is a liquid chromatogram of a sodium nitroprusside test solution obtained by a specific experiment according to the invention.
FIG. 2 is a liquid chromatogram of a sodium nitroprusside high-temperature 1 destroyed test sample solution obtained in example 3 of the present invention.
FIG. 3 is a liquid chromatogram of sodium nitroprusside obtained in example 3 of the present invention under strong light damage.
Detailed Description
The following examples are intended to illustrate the invention, but not to limit the scope of the invention.
Example 1
The instrument comprises the following steps: high performance liquid chromatography (Shimadzu, LC-2030C, UV detector).
A chromatographic column: Welchphenyl-Ether (4.6 mm. times.250 mm, 5 μm).
Mobile phase: the aqueous phase was phosphate buffer (3g tetrabutylammonium hydrogen sulfate in 1000ml of water with 2.8g anhydrous disodium hydrogen phosphate, phosphoric acid adjusted to pH 6.0); the organic phase was methanol and the elution gradient was as follows:
flow rate: 1.0 ml/min.
Column temperature: at 40 ℃.
Detection wavelength: 230 nm.
Sample introduction amount: 20 μ l.
Diluent agent: and (3) water.
Sample treatment: weighing 50mg of the product, accurately weighing, placing in a 5ml measuring flask, adding water for dissolving, quantitatively diluting with water to scale, shaking, and centrifuging to obtain test solution.
Precisely measuring the test solution, respectively injecting into a liquid chromatograph, and recording the chromatogram.
As can be seen from FIG. 3, the method of the present invention can effectively determine newly added degradation impurities in sodium nitroprusside, and can be used for quality detection of sodium nitroprusside raw materials and intermediates.
Example 2.
The instrument comprises the following steps: the same as in example 1.
A chromatographic column: ultimate XB-phenyl (3.9 mm. times.300 mm, 10 μm is suitable).
Diluent agent: the same as in example 1.
Mobile phase: the aqueous phase was phosphate buffer (2g tetrabutylammonium hydrogen sulfate in 1000ml water with 1.5g anhydrous disodium hydrogen phosphate, phosphoric acid adjusted to pH 6.0); the organic phase was acetonitrile and the elution gradient was as follows:
flow rate: 2.0 ml/min.
Column temperature: at 30 ℃.
Detection wavelength: 210 nm.
Sample introduction amount: 20 μ l.
Sample treatment: taking 8ml of sodium nitroprusside injection (specification 25mg/ml), putting the sodium nitroprusside injection into a 10ml measuring flask, quantitatively diluting the sodium nitroprusside injection to a scale with water, and shaking up to obtain a test solution; taking appropriate amount of impurity ferricyanide compound and ferrocyanide compound as reference substance, dissolving with diluent, and quantitatively diluting to obtain solution of about 40 μ g per l ml as reference substance solution.
Precisely measuring reference substance and sample solution, respectively injecting into liquid chromatograph, recording chromatogram, wherein the content of ferricyanide compound and ferricyanide compound is not more than 0.20% according to external standard method.
Example 3
Instrument, column and chromatographic conditions: the same as in example 1.
Sample treatment: strong light damage: taking sodium nitroprusside injection (specification 25mg/ml), placing for 2 hours under 5000lx illumination condition, taking out, quantitatively diluting with water to a solution containing 20mg of sodium nitroprusside in every 1ml, and shaking up to obtain a strong light test solution; high temperature 1 destruction: taking sodium nitroprusside injection (specification 25mg/ml), placing for 5 days at 60 ℃, taking out, quantitatively diluting with water to a solution containing 20mg of sodium nitroprusside in every 1ml, and shaking up to be used as a high-temperature 1 test sample solution; high temperature 2 destruction: taking sodium nitroprusside injection (specification 25mg/ml), placing the injection into high-pressure steam sterilization at 115 ℃ for 15 minutes, taking out the injection, quantitatively diluting the injection with water until each 1ml contains about 20mg of sodium nitroprusside, and shaking up the solution to be used as a high-temperature 2 test sample solution.
And (4) injecting the test solution into a liquid chromatograph respectively, and recording the chromatogram. The sodium nitroprusside injection is extremely unstable under strong light, the appearance of the sodium nitroprusside injection is changed into magenta within 2 hours, the degradation impurities are rapidly increased, and the total impurities are increased by nearly 6 percent; the sodium nitroprusside injection is placed for 5 days at the high temperature of 60 ℃, the appearance is not obviously changed, and the total impurity is increased by 0.05 percent; the sodium nitroprusside injection is sterilized for 15 minutes at the high temperature of 115 ℃, the appearance is not obviously changed, and the total impurity is increased by 0.12 percent.
Sodium nitroprusside is very sensitive to light and is relatively unstable to high temperature. Therefore, the sodium nitroprusside raw material and the preparation thereof need to be protected from light and high temperature during storage.
Claims (6)
1. A method for controlling and detecting the degradation impurities of sodium nitroprusside is characterized in that in the storage environment of sodium nitroprusside, a sample needs to adopt a high performance liquid chromatography, a chromatographic column with common phenyl bonded silica gel as a filler is used for measurement, and a mixed solution of buffer salt and an organic phase is used as a mobile phase.
2. An assay method as claimed in claim 1, wherein the storage of the sodium nitroprusside sample is: avoiding strong light and high temperature.
3. The method of claim 1, wherein the impurities that degrade sodium nitroprusside are detected by high performance liquid chromatography and an ultraviolet detector.
4. The analytical method according to claim 1, wherein a column packed with phenyl-bonded silica gel is used.
5. The method of claim 1, wherein the diluent is an aqueous solution, the concentration of sodium nitroprusside in the sample is 10-20 mg/ml, the mobile phase and the aqueous phase are phosphate buffer solutions containing 0.2-0.4% of tetrabutylammonium hydroxide and tetrabutylammonium hydrogen sulfate ion pair reagent, and the pH of the phosphoric acid is adjusted to 6.0, and the method adopts high performance liquid chromatography and ultraviolet detector to measure the ultraviolet wavelength is 210-230 nm.
6. The method for determining the impurity spectrum of the product according to any one of claims 1 to 5 through a degradation test of sodium nitroprusside raw materials and preparations: unknown degraded impurities 1, unknown degraded impurities 2, nitrite impurities, nitrate impurities, ferrocyanide compounds, ferricyanide compounds, and the like.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110707465.0A CN113390999A (en) | 2021-06-24 | 2021-06-24 | Control and detection method for sodium nitroprusside degradation impurities |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110707465.0A CN113390999A (en) | 2021-06-24 | 2021-06-24 | Control and detection method for sodium nitroprusside degradation impurities |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113390999A true CN113390999A (en) | 2021-09-14 |
Family
ID=77623854
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110707465.0A Pending CN113390999A (en) | 2021-06-24 | 2021-06-24 | Control and detection method for sodium nitroprusside degradation impurities |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113390999A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117849208A (en) * | 2023-12-07 | 2024-04-09 | 乐声药业石家庄有限公司 | Method for measuring nitrate and nitrite content in nitroglycerin tablet |
-
2021
- 2021-06-24 CN CN202110707465.0A patent/CN113390999A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117849208A (en) * | 2023-12-07 | 2024-04-09 | 乐声药业石家庄有限公司 | Method for measuring nitrate and nitrite content in nitroglycerin tablet |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Kubota et al. | Determination of benzoic acid and hippuric acid in human plasma and urine by high-performance liquid chromatography | |
| CN113588817B (en) | Method for simultaneously measuring atropine sulfate and EDTA-2Na content in atropine eye drops | |
| CN109387587B (en) | Detection method of L-2-amino-5-guanidino valeric acid enantiomer | |
| CN113390999A (en) | Control and detection method for sodium nitroprusside degradation impurities | |
| CN111983064B (en) | Detection method of aminocaproic acid and polymer in preparation thereof | |
| Michail et al. | Development and validation of a liquid chromatographic method for the determination of hydroxymethylfurfural and alpha-ketoglutaric acid in human plasma | |
| Patil et al. | Assay determination of tranexamic acid in pharmaceutical dosage form (tablet) using HPLC and ELS detector | |
| CN112903846B (en) | Analysis method for determining rivaroxaban and impurities thereof | |
| CN110441421A (en) | A kind of method of high effective liquid chromatography for measuring Entacapone tablet dissolution | |
| CN113514589B (en) | High performance liquid chromatography analysis method of stannous glucoheptonate relative substance for injection | |
| CN111380978B (en) | Method for simultaneously determining contents of coenzymes NADP and FAD in medicine | |
| CN108828110B (en) | Method for determining residual quantity of benzimidazole drugs in chicken fat | |
| Kasimedu | A Novel UPLC Method for the Estimation of Antidiabetic Drugs in Bulk and its Tablet dosage form | |
| CN112379006A (en) | Method for detecting impurity C, impurity D and impurity E in topiroxostat tablets | |
| CN113521244A (en) | Argatroban injection and preparation method thereof | |
| CN112595793B (en) | Earthworm injection detection method based on phenol determination | |
| CN114965790B (en) | Liquid phase detection method for amino acid impurities in lysine zinc gluconate particles | |
| CN112557558B (en) | Method for detecting SCH59566 impurity content in ezetimibe simvastatin tablets | |
| CN111398442A (en) | Method for detecting N- (2-nitrobenzyl) -N-methylcyclohexylamine in bromhexine hydrochloride inhalation solution | |
| CN111351886A (en) | Method for determining impurities contained in etamsylate and content of main drug thereof | |
| CN114200050B (en) | HPLC detection method for content of related substances in p-bromoanisole | |
| CN113075306B (en) | Method for detecting content of edetate disodium in pantoprazole sodium | |
| CN111380991B (en) | Method for detecting content of degradation impurities in vitamin C medicament | |
| CN117890496B (en) | Method for detecting related substances of compound preparation of novel oral solution of guaifenesin | |
| CN111812227A (en) | Method for analyzing benserazide impurity A in poly-barserazide compound preparation |
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 |