CN106404949A - Detection method for content of carprofen - Google Patents
Detection method for content of carprofen Download PDFInfo
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- CN106404949A CN106404949A CN201610784145.4A CN201610784145A CN106404949A CN 106404949 A CN106404949 A CN 106404949A CN 201610784145 A CN201610784145 A CN 201610784145A CN 106404949 A CN106404949 A CN 106404949A
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- 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
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Abstract
The invention provides a detection method for the content of carprofen. According to the invention, an Agilent-1260 high-performance liquid chromatograph is used for analysis; a DAD detector is used; detection wavelength is 235 nm; and a sample size is 20 [mu]l. The detection method comprises a step of carrying out determination with the Agilent-1260 high-performance liquid chromatograph wondasil C18, wherein chromatographic conditions are that a chromatographic column uses octadecylsilane-bonded silica gel as a filler; a mobile phase is methanol-sodium dihydrogen phosphate with a concentration of 0.02 mol/l and the volume fraction of methanol is 65 to 80%; flow velocity is 0.6 to 1.5 ml/min; and column temperature is 25 to 40 DEG C. The detection method for the content of carprofen realizes analysis and detection via liquid chromatography, and is simple, convenient, good in separating effect, high in precision and accuracy, accurate and reliable in detection results, great in detection scope, and high in sensitivity and accuracy.
Description
Technical field
The invention belongs to medicament contg detection field, especially relate to a kind of detection method of content of ibuprofen.
Background technology
NSAID (non-steroidal anti-inflammatory drug) is the anti-inflammatory agent that a class does not contain steroidal structure, is clinically widely used in mammal galactophore
Inflammation, the alleviation of metritiss, arthritic treatment and various heating and pain symptom, due to evident in efficacy, have become consumption at present
One of most primary species.
Carprofen is 2- aryl propionic non-steroid anti-inflammatory analgesic, is mainly used in joint of animal treatment, with other non-steroids
Body anti-inflammatory agent is compared, and effect is substantially, gastrointestinal tract acts on and Nephrotoxicity is many smaller.
Carprofen, English name：Carprofen, No. CAS is 53716-49-7, and molecular formula is C15H12CLNO2, molecular weight
For 273.71, fusing point is 197-198 DEG C, structural formula as shown in (I),
The method of monitoring carprofen is Solid-Phase Extraction LC-MS technology at present, and equipment is complicated, and process is complicated, to operator
Member's skill set requirements are higher, are not suitable for industrial mass sample operations.
Content of the invention
In view of this, it is contemplated that proposing a kind of detection method of content of ibuprofen, testing result accurately and reliably, detects model
Enclose big.
For reaching above-mentioned purpose, the technical scheme is that and be achieved in that：
A kind of detection method of content of carprofen, is analyzed using Agilent-1260 high performance liquid chromatograph, adopts
DAD detector, Detection wavelength is 235nm, and sample size is 20 μ l, comprises the steps：
(1) chromatographic condition：
Chromatographic column is with octadecylsilane chemically bonded silica as implant, using Agilent-1260 high performance liquid chromatography
wondasil C18It is measured；Mobile phase is methanol -0.02mol/l sodium dihydrogen phosphate, and wherein, the volume fraction of methanol is 65-
80%；Flow velocity is 0.6-1.5ml/min；Column temperature is 25-40 DEG C；
(2) sample treatment：
Precision weighing standard substance about 50.0mg, puts in 50ml volumetric flask, is settled to scale after adding the dissolving of chromatograph methanol
Line, uses the organic filter of 0.45m to filter after shaking up；
The test sample of precision weighing equal in quality, puts in 50ml volumetric flask, is settled to quarter after adding the dissolving of chromatograph methanol
Degree line, uses the organic filter of 0.45m to filter after shaking up；
(3) algoscopy：By above-mentioned condition, respectively auto injection is carried out to standard substance and test sample, with external standard method with peak face
The long-pending content calculating carprofen.
Further, the mensure formula of the calculating carprofen content in described step (3) is：
Carprofen content (%)=(product to be tested peak area/standard substance peak area) * (standard concentration/product to be tested concentration) *
100%.
Further, the high performance liquid chromatography wondasil C in described step (1)18Post is 4.6*250mm, 5 μm
wondasil C18Post, 4.6*150mm, 5 μm of wondasil C18Post or 4.6*100mm, 5 μm of wondasil C18In post one
Kind.
Preferably, in described step (1), the volume fraction of methanol is 75%.
Preferably, in described step (1), flow velocity is 1ml/min.
Preferably, in described step (1), column temperature is 30 DEG C.
With respect to prior art, the detection method of content of carprofen of the present invention has the advantage that：
The detection method of content of carprofen of the present invention is analyzed detecting by liquid chromatography, is a kind of letter
Just, the detection method of the high carprofen content of good separating effect, preci-sion and accuracy, testing result accurately and reliably, detects model
Enclose big, sensitivity is high and accuracy is high.
Brief description
The accompanying drawing constituting the part of the present invention is used for providing a further understanding of the present invention, the schematic reality of the present invention
Apply example and its illustrate, for explaining the present invention, not constituting inappropriate limitation of the present invention.In the accompanying drawings：
Fig. 1 is the HPLC detection collection of illustrative plates of the carprofen standard substance described in the embodiment of the present invention；
Fig. 2 is the linear relationship chart of carprofen and regression equation described in the embodiment of the present invention.
Specific embodiment
Unless otherwise indicated, term used herein is respectively provided with the conventional implication understanding of those skilled in the art, in order to
Readily appreciate the present invention, some terms used herein have been carried out following definitions.
To describe the present invention with reference to embodiment in detail.
Embodiment 1
Precision weighs standard substance 50.0mg, is placed in 50ml volumetric flask, dilutes and be settled to after additive color spectrum methanol ultrasonic dissolution
Graduation mark, takes solution to filter through 0.45 μm of disposably organic filter with 1ml syringe, obtains standard solution；Precision weighs phase
With the test sample of weight, it is placed in 50ml volumetric flask, dilution after additive color spectrum methanol ultrasonic dissolution is settled to graduation mark, is noted with 1ml
Emitter takes solution to filter through 0.45 μm of disposably organic filter, obtains need testing solution.Standard solution and need testing solution are each
5 parts of configuration.
Chromatographic condition：With Agilent-1260 high performance liquid chromatography wondasil C18Post (4.6*250mm, 5 μm)；Flowing
Phase：Methanol -0.02mol/l sodium dihydrogen phosphate (75:25)；Flow velocity：1ml/min；Detection wavelength：235nm；Acquisition time 25min；
Detector：UV-detector；Column temperature：30℃.
By above-mentioned chromatographic condition, to all standard substance and test sample sample introduction respectively, as shown in figure 1,5 samples of standard solution
Sample introduction gained peak area meansigma methodss are 0.16% for 5969, RSD value to product respectively；5 samples of need testing solution sample introduction gained respectively
Peak area meansigma methodss are 0.23% for 5932, RSD value.
According to formula：
Carprofen content (%)=(product to be tested peak area/standard substance peak area) * (standard concentration/product to be tested concentration) *
100%
Using external standard method by standard substance and test sample meansigma methodss substitute into the content of test sample carprofen is
99.51%.
The methodological study of detection method described in the embodiment of the present invention 1
(1) precision is investigated
Weigh same carprofen sample, carry out 6 times repeating to test, peak area table 1；
Table 1 carprofen repeats the area tested for 6 times
| Number of times | 1 | 2 | 3 | 4 | 5 | 6 |
| Peak area | 5980 | 5990 | 5972 | 5978 | 5964 | 5997 |
Peak area RSD is 0.20%, and result shows, this HPLC method detection carprofen content precision is high, favorable reproducibility.
(2) linear relationship test
According to invention methods described, the storing solution being 10mg/ml with the volumetric flask compound concentration of 50ml, plus methanol are dilute respectively
It is interpreted into the standard solution of 0.1mg/ml, 0.2mg/ml, 0.5mg/ml, 1.0mg/ml, 1.5mg/ml, 2.0mg/ml, according to this
Bright analysis method, carries out data regression to sample size (X) and peak area (Y), and linear relationship is as shown in Fig. 2 regression equation
For y=6033.8x-20.68.Result shows carprofen in 2ug-40ug concentration range, and peak area and sample size are in good
Linear relationship.
(3) it is loaded recovery experiment
The carprofen sample accurately weighing known content is appropriate, more a certain amount of carprofen sample of accurate addition respectively
80%th, 100%, 120%, make the carprofen concentration in need testing solution respectively in the high, medium and low region of carprofen standard curve
Each 3 parts, by the operation under item of the present invention, measure in accordance with the law, calculate the response rate, result average recovery rate for 99.68%, RSD is
0.45%, show that the method average recovery is good.
To sum up show detection method of content good stability of the present invention, precision good it is adaptable to carprofen containing measurement
Fixed.
Precision weighing 50mg same sample is placed in 50ml volumetric flask again, is settled to scale after additive color spectrum methanol dilution
Line；Precision weighing 20,40,50,60,80mg standard substance be respectively placed in 50ml volumetric flask, be settled to after additive color spectrum methanol dilution
Graduation mark, obtains standard solution 0.4mg/ml, 0.8mg/ml, 1.0mg/ml, 1.2mg/ml, 1.6mg/ml.
Chromatographic condition：Agilent Zorbax Extend C18Chromatographic column (3*50mm, 3 μm), column temperature：30 DEG C, flow velocity：
0.8ml/min；Detection wavelength：235nm；Sample size：10 μ l, mobile phase：A methanol, B 0.2% aqueous formic acid, using gradient
The method rinsed see table 2.
Table 2 gradient washes
| Time (min) | A | B |
| 0 | 10 | 90 |
| 0.5 | 10 | 90 |
| 1.5 | 90 | 10 |
| 4 | 90 | 10 |
| 4.5 | 10 | 90 |
| 5.5 | 10 | 90 |
Mass Spectrometry Conditions：Electron spray ionisation source (ESI)；Spray voltage (IS) -4.2kv；Multiple-reaction monitoring pattern (MRM)；Negative
Ion scan；Atomization gas pressure (GSI) 50psi, secondary air speed (GS2) 55L/min, gas curtain atmospheric pressure (GUR) 20psi, from
150 DEG C of source temperature (TEM), collides chamber pressure (CAD) 7psi, and selective enumeration method charge-mass ratio (m/z) is 193 negatively charged dividing
Daughter ion peak.
By above-mentioned condition, to standard substance 1,2,3,4,5 and test sample sample introduction respectively, record the content of carprofen in test sample
For 99.3%.
Embodiment 2
Sample and standard substance configure with embodiment 1.
Chromatographic condition：With Agilent-1260 high performance liquid chromatography wondasil C18Post (4.6*150mm, 5 μm)；Flowing
Phase：Methanol -0.02mol/l sodium dihydrogen phosphate (65:35)；Flow velocity：0.6ml/min；Detection wavelength：235nm；Acquisition time
35min；Column temperature：40℃.
By above-mentioned chromatographic condition, to all standard substance and test sample sample introduction respectively, as shown in figure 1,5 samples of standard solution
Sample introduction gained peak area meansigma methodss are 0.34% for 6014, RSD value to product respectively；5 samples of need testing solution sample introduction gained respectively
Peak area meansigma methodss are 0.43% for 5946, RSD value.
According to formula：
Carprofen content (%)=(product to be tested peak area/standard substance peak area) * (standard concentration/product to be tested concentration) *
100%
Using external standard method by standard substance and test sample meansigma methodss substitute into the content of test sample carprofen is
99.02%.
Embodiment 3
Sample and standard substance configure with embodiment 1.
Chromatographic condition：With Agilent-1260 high performance liquid chromatography wondasil C18Post (4.6*100mm, 5 μm)；Flowing
Phase：Methanol -0.02mol/l sodium dihydrogen phosphate (80:20)；Flow velocity：1.5ml/min；Detection wavelength：235nm；Acquisition time
25min；Column temperature：25℃.
By above-mentioned chromatographic condition, to all standard substance and test sample sample introduction respectively, as shown in figure 1,5 samples of standard solution
Sample introduction gained peak area meansigma methodss are 0.47% for 6203, RSD value to product respectively；5 samples of need testing solution sample introduction gained respectively
Peak area meansigma methodss are 0.63% for 6016, RSD value.
According to formula：
Carprofen content (%)=(product to be tested peak area/standard substance peak area) * (standard concentration/product to be tested concentration) *
100%
Using external standard method by standard substance and test sample meansigma methodss substitute into the content of test sample carprofen is
100.14%.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Within god and principle, any modification, equivalent substitution and improvement made etc., should be included within the scope of the present invention.
Claims (6)
1. a kind of detection method of content of carprofen it is characterised in that：Carried out using Agilent-1260 high performance liquid chromatograph
Analysis, using DAD detector, Detection wavelength is 235nm, and sample size is 20 μ l, comprises the steps：
(1) chromatographic condition：
Chromatographic column is with octadecylsilane chemically bonded silica as implant, using Agilent-1260 high performance liquid chromatography
wondasil C18It is measured；Mobile phase is methanol -0.02mol/l sodium dihydrogen phosphate, and wherein, the volume fraction of methanol is 65-
80%；Flow velocity is 0.6-1.5ml/min；Column temperature is 25-40 DEG C；
(2) sample treatment：
Precision weighing standard substance about 50.0mg, puts in 50ml volumetric flask, is settled to graduation mark, shakes after adding the dissolving of chromatograph methanol
The organic filter of 0.45m is used to filter after even；
The test sample of precision weighing equal in quality, puts in 50ml volumetric flask, is settled to graduation mark after adding the dissolving of chromatograph methanol,
The organic filter of 0.45m is used to filter after shaking up；
(3) algoscopy：By above-mentioned condition, respectively auto injection is carried out to standard substance and test sample, with external standard method in terms of peak area
Calculate the content of carprofen.
2. carprofen according to claim 1 detection method of content it is characterised in that：Meter in described step (3)
Calculate carprofen content mensure formula be：
Carprofen content (%)=(product to be tested peak area/standard substance peak area) * (standard concentration/product to be tested concentration) *
100%.
3. carprofen according to claim 1 detection method of content it is characterised in that：Height in described step (1)
Effect liquid phase chromatogram wondasil C18Post is 4.6*250mm, 5 μm of wondasil C18Post, 4.6*150mm, 5 μm of wondasil
C18Post or 4.6*100mm, 5 μm of wondasil C18One of post.
4. carprofen according to claim 1 detection method of content it is characterised in that：Methanol in described step (1)
Volume fraction be 75%.
5. carprofen according to claim 1 detection method of content it is characterised in that：Flow velocity in described step (1)
For 1ml/min.
6. carprofen according to claim 1 detection method of content it is characterised in that：Column temperature in described step (1)
Spend for 30 DEG C.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610784145.4A CN106404949A (en) | 2016-08-31 | 2016-08-31 | Detection method for content of carprofen |
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|---|---|---|---|
| CN201610784145.4A CN106404949A (en) | 2016-08-31 | 2016-08-31 | Detection method for content of carprofen |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112526038A (en) * | 2020-12-15 | 2021-03-19 | 河北科技大学 | Carprofen and related substance detection method |
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| CN103278594A (en) * | 2013-04-24 | 2013-09-04 | 宁波检验检疫科学技术研究院 | Universal rapid detection method for micromolecule poisonous and harmful materials in powdery food |
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2016
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101650345A (en) * | 2009-08-31 | 2010-02-17 | 山东京博控股发展有限公司 | Analysis method of content of 2-chloronicotinic acid |
| CN103278594A (en) * | 2013-04-24 | 2013-09-04 | 宁波检验检疫科学技术研究院 | Universal rapid detection method for micromolecule poisonous and harmful materials in powdery food |
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| NABILA BOUCHAIR等: "Reversed-phase high-performance liquid chromatographic separation of some 2-arylpropionic acids using vancomycin as chiral stationary phase", 《JOURNAL OF IRANIAN CHEMICAL SOCIETY》 * |
| PASQUALE GALLO 等: "Confirmatory analysis of non-steroidal anti-inflammatory drugs in bovine milk by high-performance liquid chromatography with fluorescence detection", 《JOURNAL OF CHROMATOGRAPHY A》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN112526038A (en) * | 2020-12-15 | 2021-03-19 | 河北科技大学 | Carprofen and related substance detection method |
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Application publication date: 20170215 |
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