CN106248819A - The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel - Google Patents
The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel Download PDFInfo
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- CN106248819A CN106248819A CN201610562816.2A CN201610562816A CN106248819A CN 106248819 A CN106248819 A CN 106248819A CN 201610562816 A CN201610562816 A CN 201610562816A CN 106248819 A CN106248819 A CN 106248819A
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- sodium hyaluronate
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- 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
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- 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
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- 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
- G01N2030/042—Standards
- G01N2030/045—Standards internal
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Abstract
The present invention relates to the detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel.Method particularly includes: product is hydrolyzed through hyaluronidase, through filtering after completing, uses high performance liquid chromatograph to carry out assay.The present invention is the content detection of lidocaine hydrochloride in medical cross-linking sodium hyaluronate, makes product hydrolysis by hyaluronidase, and adds mobility, mix homogeneously, membrane filtration, then high performance liquid chromatograph detection.Product hydrolysis i.e. decreases the hyaluronate sodium impact on detection, improves Detection accuracy, easy to operate, reproducible.
Description
Technical field
The present invention relates to medicine detection technique field, particularly to hydrochloric acid profit in a kind of medical cross-linking sodium hyaluronate gel
The detection method of many caines content
Background technology
Lidocaine hydrochloride content detection in medical cross-linking sodium hyaluronate gel, wherein medical cross-linking sodium hyaluronate coagulates
Glue is particulate matter and lubricating fluid composition, and wherein particulate matter is close tridimensional network, and lidocaine hydrochloride is dissolved in product
In, may exist in network structure.Lidocaine hydrochloride content detection, product needed filters, and possibly cannot make all of
Hydrochloric acid benefit cartoon filter membrane, affects accuracy and the repeatability of testing result.
Summary of the invention
The technical problem to be solved is to exist for existing Chinese Pharmacopoeia lidocaine hydrochloride content detection
Deficiency and the detection method of lidocaine hydrochloride content in a kind of preparation is provided, the method is simple to operate, and accuracy is good, repeat
Property high
The technical problem to be solved can be achieved through the following technical solutions:
The detection method of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel, has following steps:
(1) flowing phase preparation steps
Take 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, be diluted with water to
1000ml, phosphate solution: acetonitrile=500:500, regulation pH is 8.0；
(2) reference substance solution preparation steps
Take lignocaine reference substance appropriate, accurately weighed, with flowing phase dilution to 0.2mg/ml～1.0mg/ml；
(3) need testing solution preparation steps
Take the medical cross-linking sodium hyaluronate gel 1.0g of hydrochloric benefit card, be the hyalomitome of 20-100U/ml by concentration
Acid sodium enzymatic solution 3.0-5.0ml, constant temperature 12-28 hour in 37 DEG C of water-baths, it is settled to 10ml, 0.45 μm mutually with flowing after dissolving
Membrane filtration, standby；
(4) testing procedure:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
The each 20 μ L of reference substance solution that precision takes need testing solution prepared by step (3), prepared by step (2) inject liquid phase colors
Spectrometer, records chromatogram, with external standard method by calculated by peak area content.
In a preferred embodiment of the invention, in described step (2), described reference substance solution concentration is 0.3mg/ml-
0.4mg/ml。
In a preferred embodiment of the invention, in described step (3), the concentration of described hyaluronidase solution is 50U/
ml-60U/ml。
In a preferred embodiment of the invention, in described step (3), in 37 DEG C of water-baths, constant temperature is 20-24 hour.
The present invention makes medical cross-linking sodium hyaluronate gel hydrolyze by hyaluronidase, then filters, and makes hydrochloric acid profit
Many caines, completely by filter membrane, are filled in detection sample, and detection correctness is good, and repeatability is high, easy to operate.
Accompanying drawing explanation
Fig. 1 is the liquid chromatogram of the reference substance solution of the embodiment of the present invention 1.
Fig. 2 is the liquid chromatogram of the need testing solution of the embodiment of the present invention 1.
Fig. 3 is the liquid chromatogram of the reference substance solution of the embodiment of the present invention 2.
Fig. 4 is the liquid chromatogram of the need testing solution of the embodiment of the present invention 2.
Fig. 5 is the liquid chromatogram of the reference substance solution of the embodiment of the present invention 3.
Fig. 6 is the liquid chromatogram of the need testing solution of the embodiment of the present invention 3.
Fig. 7 is the liquid chromatogram of the reference substance solution of the embodiment of the present invention 4.
Fig. 8 is the liquid chromatogram of the need testing solution of the embodiment of the present invention 4.
Fig. 9 is the liquid chromatogram of the reference substance solution of the embodiment of the present invention 5.
Figure 10 is the liquid chromatogram of the need testing solution of the embodiment of the present invention 5.
Detailed description of the invention:
Below in conjunction with embodiment, the present invention is further described, and those skilled in the art can be helped to be more completely understood by
The present invention.But limit the present invention never in any form, the equivalent of all any this areas done according to present disclosure
Within belonging to protection scope of the present invention.
Embodiment 1
Flowing preparation mutually: 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, uses
Water is diluted to 1000ml, phosphate solution: acetonitrile=500:500, and regulation pH is 8.0
Reference substance solution is prepared: take lignocaine reference substance 15.6mg, with flowing phase dilution to 0.312mg/ml, standby.
Need testing solution is prepared: take product 1.1014g, with the hyaluronate sodium enzymatic solution 4.0ml that concentration is 60U/ml, and 37
Constant temperature 24 hours in DEG C water-bath, is settled to 10ml mutually with flowing after dissolving, and 0.45 μm membrane filtration is standby.
Detection:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
Precision takes need testing solution, each 20 μ L of reference substance solution inject chromatograph of liquid, records chromatogram such as Fig. 1 and Fig. 2,
With external standard method by calculated by peak area content.The results are shown in Table 1
Table 1
| Sample ID | Retention time | Peak area % | Peak area | Remarks |
| JJ-10 | 13.852 | 100.00 | 1843474 | Reference substance |
| JJ-2 | 13.814 | 100.00 | 1775824 | Test sample |
Embodiment 2
Flowing preparation mutually: 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, uses
Water is diluted to 1000ml, phosphate solution: acetonitrile=500:500, and regulation pH is 8.0
Reference substance solution is prepared: take lignocaine reference substance 15.1mg, with flowing phase dilution to about 0.302mg/ml, standby
With.
Need testing solution is prepared: take product 1.0689g, with the hyaluronate sodium enzymatic solution 4.0ml that concentration is 50U/ml, and 37
Constant temperature 24 hours in DEG C water-bath, is settled to 10ml mutually with flowing after dissolving, and 0.45 μm membrane filtration is standby.
Detection:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
Precision takes need testing solution, each 20 μ L of reference substance solution inject chromatograph of liquid, records chromatogram such as Fig. 3 and Fig. 4,
With external standard method by calculated by peak area content.Result sees table 2
Table 2
| Sample ID | Retention time | Peak area % | Peak area | Remarks |
| JJ-11 | 13.842 | 100.00 | 1833534 | Reference substance |
| JJ-4 | 13.776 | 100.00 | 1786631 | Test sample |
Embodiment 3
Flowing preparation mutually: 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, uses
Water is diluted to 1000ml, phosphate solution: acetonitrile=500:500, and regulation pH is 8.0
Reference substance solution is prepared: take lignocaine reference substance 16.6mg, with flowing phase dilution to about 0.332mg/ml, standby
With.
Need testing solution is prepared: take product 1.1826g, with the hyaluronate sodium enzymatic solution 4.0ml that concentration is 100U/ml,
Constant temperature 16 hours in 37 DEG C of water-baths, is settled to 10ml mutually with flowing after dissolving, and 0.45 μm membrane filtration is standby.
Detection:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
Precision takes need testing solution, each 20 μ L of reference substance solution inject chromatograph of liquid, records chromatogram such as Fig. 5 and Fig. 6,
With external standard method by calculated by peak area content.Result sees table 3
Table 3
| Sample ID | Retention time | Peak area % | Peak area | Remarks |
| JJ-1 | 12.952 | 100.00 | 1943474 | Reference substance |
| JJ-10 | 13.852 | 100.00 | 1885474 | Test sample |
Embodiment 4
Flowing preparation mutually: 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, uses
Water is diluted to 1000ml, phosphate solution: acetonitrile=500:500, and regulation pH is 8.0
Reference substance solution is prepared: take lignocaine reference substance 15.3mg, with flowing phase dilution to about 0.306mg/ml, standby
With.
Need testing solution is prepared: take product 1.0307g, is settled to 10ml mutually with flowing, and 0.45 μm membrane filtration is standby.
Detection:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
Precision takes need testing solution, each 20 μ L of reference substance solution inject chromatograph of liquid, records chromatogram such as Fig. 7 and Fig. 8,
With external standard method by calculated by peak area content.Result sees table 4
Table 4
| Sample ID | Retention time | Peak area % | Peak area | Remarks |
| JJ-5 | 13.525 | 100.00 | 1753475 | Reference substance |
| JJ-1 | 13.853 | 100.00 | 614346 | Test sample |
Embodiment 5
Flowing preparation mutually: 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, uses
Water is diluted to 1000ml, phosphate solution: acetonitrile=500:500, and regulation pH is 8.0
Reference substance solution is prepared: take lignocaine reference substance 15.1mg, with flowing phase dilution to about 0.302mg/ml, standby
With.
Need testing solution is prepared: take product 1.0204g, with the hyaluronate sodium enzymatic solution 4.0ml that concentration is 20U/ml, and 37
Constant temperature 28 hours in DEG C water-bath, is settled to 10ml mutually with flowing after dissolving, and 0.45 μm membrane filtration is standby.
Detection:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
Precision takes need testing solution, each 20 μ L of reference substance solution inject chromatograph of liquid, records chromatogram such as Fig. 9 and Tu
10, with external standard method by calculated by peak area content.Result sees table 5
Table 5
| Sample ID | Retention time | Peak area % | Peak area | Remarks |
| JJ-1 | 13.832 | 100.00 | 1743447 | Reference substance |
| JJ-1 | 12.194 | 100.00 | 584759 | Test sample |
By above example gained finished product censorship, assay and process parameter are summarized as follows table 6:
Table 6
By upper table it could be assumed that, embodiment 1, embodiment 2, embodiment 3 all use hyaluronidase to be hydrolyzed after examine
Surveying, result is close to theoretical value, and result is the most stable, reproducible, and relative standard deviation is 0.3%.Embodiment 4, embodiment 5 does not has
There are employing hyaluronic acid enzyme hydrolysis, Lower result, poor repeatability, relative standard deviation 8.0%.
Claims (4)
1. the detection method of lidocaine hydrochloride content in a medical cross-linking sodium hyaluronate gel, it is characterised in that have
Following steps:
(1) flowing phase preparation steps
Take 1mol/L sodium dihydrogen phosphate 1.3ml and 0.5mol/L disodium phosphate soln 32.5ml, be diluted with water to
1000ml, phosphate solution: acetonitrile=500:500, regulation pH is 8.0；
(2) reference substance solution preparation steps
Take lignocaine reference substance appropriate, accurately weighed, with flowing phase dilution to 0.2mg/ml～1.0mg/ml；
(3) need testing solution preparation steps
Take the medical cross-linking sodium hyaluronate gel 1.0g of hydrochloric benefit card, be the hyaluronate sodium of 20-100U/ml by concentration
Enzymatic solution 3.0-5.0ml, constant temperature 12-28 hour in 37 DEG C of water-baths, it is settled to 10ml, 0.45 μm filter membrane mutually with flowing after dissolving
Filter, standby；
(4) testing procedure:
Instrument: high performance liquid chromatograph, octadecylsilane chemically bonded silica chromatographic column
Column temperature: 25 DEG C, flow velocity: 1ml/min
The each 20 μ L of reference substance solution that precision takes need testing solution prepared by step (3), prepared by step (2) inject liquid chromatograph
Instrument, records chromatogram, with external standard method by calculated by peak area content.
The detection side of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel the most as claimed in claim 1
Method, it is characterised in that in described step (2), described reference substance solution concentration is 0.3mg/ml-0.4mg/ml.
The detection side of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel the most as claimed in claim 1
Method, it is characterised in that in described step (3), the concentration of described hyaluronidase solution is 50U/ml-60U/ml.
The detection side of lidocaine hydrochloride content in a kind of medical cross-linking sodium hyaluronate gel the most as claimed in claim 1
Method, it is characterised in that in described step (3), in 37 DEG C of water-baths, constant temperature is 20-24 hour.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111208236A (en) * | 2020-03-05 | 2020-05-29 | 遂成药业股份有限公司 | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography |
| RU2725157C1 (en) * | 2019-12-26 | 2020-06-30 | Федеральное государственное бюджетное учреждение науки Институт общей и неорганической химии им. Н.С. Курнакова Российской академии наук (ИОНХ РАН) | Ion-selective electrode membrane for determining lidocaine |
| CN112410398A (en) * | 2020-12-07 | 2021-02-26 | 海雅美生物技术(珠海)有限公司 | Sterility detection method for crosslinked sodium hyaluronate gel for injection |
| CN113504321A (en) * | 2021-06-29 | 2021-10-15 | 杭州协合医疗用品有限公司 | Method for simultaneously detecting residual cleaning agent n-hexane and ethyl acetate |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2725157C1 (en) * | 2019-12-26 | 2020-06-30 | Федеральное государственное бюджетное учреждение науки Институт общей и неорганической химии им. Н.С. Курнакова Российской академии наук (ИОНХ РАН) | Ion-selective electrode membrane for determining lidocaine |
| CN111208236A (en) * | 2020-03-05 | 2020-05-29 | 遂成药业股份有限公司 | Method for measuring related substances of lidocaine hydrochloride by high performance liquid chromatography |
| CN112410398A (en) * | 2020-12-07 | 2021-02-26 | 海雅美生物技术(珠海)有限公司 | Sterility detection method for crosslinked sodium hyaluronate gel for injection |
| CN113504321A (en) * | 2021-06-29 | 2021-10-15 | 杭州协合医疗用品有限公司 | Method for simultaneously detecting residual cleaning agent n-hexane and ethyl acetate |
| CN113504321B (en) * | 2021-06-29 | 2023-09-01 | 杭州协合医疗用品有限公司 | Method for simultaneously detecting residual cleaning agent n-hexane and ethyl acetate |
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Application publication date: 20161221 |