CN110221003A - The remaining detection method of ethyl alcohol in a kind of hemostatic material - Google Patents
The remaining detection method of ethyl alcohol in a kind of hemostatic material Download PDFInfo
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- CN110221003A CN110221003A CN201910650620.2A CN201910650620A CN110221003A CN 110221003 A CN110221003 A CN 110221003A CN 201910650620 A CN201910650620 A CN 201910650620A CN 110221003 A CN110221003 A CN 110221003A
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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
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Abstract
The invention discloses the remaining detection methods of ethyl alcohol in a kind of hemostatic material, belong to medical material chemical residual detection technique field.The remaining detection method of ethyl alcohol is to extract ethyl alcohol in hemostatic material as internal standard using normal propyl alcohol in hemostatic material of the invention, measures ethanol content with gas chromatography.Go out peak position in the method for the present invention and ethyl alcohol appearance positional distance is moderate, meet separating degree requirement, peak shape is good;Testing result is accurate, and stability is high.
Description
Technical field
The invention belongs to medical material chemical residual detection technique fields, and in particular to ethyl alcohol residual in a kind of hemostatic material
Detection method.
Background technique
Ethyl alcohol used in the production technology of hemostatic material is difficult to vapor away completely, and the presence of micro ethanol will make in product
It generates different degrees of stimulate the reaction to patient in use, may also increase the organs such as liver in addition to generating pain
Disease risks, so answering the residual quantity of strict control ethyl alcohol.In pharmacopeia clear stipulaties Residual ethanol must not be higher than 0.5%, but
Be Residual ethanol in pharmacopeia detection method it is cumbersome, Check-Out Time is longer.
Using Residual ethanol in external standard method detection preparation in existing patent CN201510988447.9, although method is simple
It is single, but external standard method, for volatile ethyl alcohol, precision and stability all significantly limit its testing result.Patent
Dimethyl sulfoxide and dimethylformamide (DMF) are used in CN201711099859.2 and patent CN201310175426.6 respectively
For solvent, increase experiment risk, temperature programming range is wide, and time-consuming.Use acetonitrile as molten in patent CN201510651237.0
Agent, and prepare liquid is obtained with organic membrane filtration, the volatilization time of ethyl alcohol is increased, keeps testing result less than normal.
Summary of the invention
The present invention is provided a method with regard to the remaining detection of hemostatic material ethyl alcohol.Its chief value is to improve existing medicine
Limitation existing for Residual ethanol detection method in allusion quotation, while having evaded after hemostatic material imbibition expansion that volume is uncertain asks
Topic, the method for the invention is simple and easy, and assay accuracy is high.
In order to achieve the above object, the present invention adopts the following technical scheme:
Technical scheme is as follows:
The detection method of Residual ethanol, includes the following steps: in a kind of hemostatic material
(1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol, with ultrapure water constant volume, shakes up, is prepared into concentration
For 0.25%~2% solution, as internal standard stock solution;
(2) preparation method of standard solution: precision measures a certain amount of ethyl alcohol and is shaken up with ultrapure water constant volume, is prepared into dense
The solution that degree is 5%, as ethyl alcohol Standard Stock solutions;Respectively measure ethyl alcohol Standard Stock solutions in right amount and a certain amount of internal standard
Stock solution is configured to the series standard solution that internal standard concentration is identical, concentration of alcohol is 0.0025%~0.5%;
(3) preparation method of test solution: precision weighs appropriate test sample, adds a certain amount of internal standard stock solution, uses
Ultrapure water constant volume keeps internal standard concentration identical as internal standard concentration in standard items;5~60min of extraction is shaken up, test solution is obtained;
(4) determination step:
Chromatographic condition:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);
Injector temperature: 200 DEG C;Detector: 220 DEG C of FID, temperature;
Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;
Input mode: head space split sampling, split ratio 10:1~20:1;
Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;
Air: 400mL/min, hydrogen: 30mL/min, make-up gas (N2): 25mL/min;
Test solution headspace sampling prepared by step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
On the basis of above scheme, in the step (4), using head space split sampling, split ratio 10:1.
On the basis of above scheme, in the step (1), the concentration of the internal standard stock solution of preparation is 0.5%.
On the basis of above scheme, in the step (2), the series standard solution of preparation be internal standard concentration 0.01%,
The serial solution that concentration of alcohol is 0.005%~0.2%.
The advantages of technical solution of the present invention:
Inventor had once attempted a variety of internal standard solvents and had detected, but found in actual mechanical process, acetone, methylene chloride
Etc. a variety of internal standard compounds, separating degree, appearance time etc. cannot be optimal effect.It is that internal standard is molten that the present invention, which selects normal propyl alcohol,
Liquid, peak position and ethyl alcohol appearance positional distance are moderate out, have not only met separating degree requirement, but also will not generate error because of hypertelorism,
The present invention uses split sampling, reduces hangover, preferably controls peak shape;Temperature programming range is controlled, detection time is reduced;To confession
Test product is extracted, and the ethyl alcohol in test sample is completely dissolved in ultrapure water, increases inspection result accuracy.
In addition, the confession of stable homogeneous cannot be made due to the special imbibition of hemostatic material and property not soluble in water
Hemostatic material is soaked in ultrapure water by test sample solution using ethyl alcohol and water arbitrarily than the property dissolved each other, and is sufficiently extracted, and is obtained
Test solution, can both solve that hemostatic material is not soluble in water and the property of water swelling, can also Accurate Determining wherein ethyl alcohol remains.
Detailed description of the invention
The measurement of Fig. 1 ethyl alcohol and normal propyl alcohol retention time, wherein ordinate is response, and abscissa is retention time;
The standard curve of Fig. 2 ethyl alcohol, wherein ordinate is the ratio between ethyl alcohol and normal propyl alcohol peak area, and abscissa is that ethyl alcohol is dense
Degree.
Specific embodiment
Term as used in the present invention generally has those of ordinary skill in the art usual unless otherwise specified
The meaning of understanding.
Combined with specific embodiments below, and referring to the data further detailed description present invention.Following embodiment only be
It illustrates the present invention, rather than limits the scope of the invention in any way.
Embodiment 1
1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol solution in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 0.5% internal standard stock solution.
2) preparation of standard solution
Method: precision measures a certain amount of ethyl alcohol in 100mL volumetric flask, with ultrapure water constant volume, shakes up, obtains 5% second
Alcohol standard solution.Ethyl alcohol standard solution is measured respectively in right amount with a certain amount of internal standard stock solution, and being configured to internal standard concentration is
0.02%, concentration of alcohol is the series standard solution of 0.005%-0.2%.
3) preparation method of test solution: precision weighs 0.5g sample in 100mL volumetric flask, adds a certain amount of internal standard
Stock solution, with ultrapure water constant volume, internal standard solution concentration is 0.02%, shakes up to obtain test solution.
4) determination step:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);Injector temperature: 200 DEG C;Detector: FID, temperature 220
℃;Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;Sample introduction
Mode: head space split sampling, split ratio 10:1;Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;Air: 400mL/min, hydrogen
Gas: 30mL/min, make-up gas (N2): 25mL/min.
Test solution headspace sampling prepared by step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
Embodiment 2
1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol solution in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 0.5% internal standard stock solution
2) preparation method of standard solution: precision measures a certain amount of ethyl alcohol in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 5% ethyl alcohol standard solution.Respectively measure ethyl alcohol standard solution in right amount and a certain amount of internal standard stock solution,
It is configured to the series standard solution that internal standard concentration is 0.01%, concentration of alcohol is 0.005%-0.2%.
3) preparation method of test solution: precision weighs 0.25g sample in 100mL volumetric flask, adds in a certain amount of
Stock solution is marked, with ultrapure water constant volume, internal standard solution concentration is 0.01%, shakes up to obtain test solution.
4) determination step:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);Injector temperature: 200 DEG C;Detector: FID, temperature 220
℃;Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;Sample introduction
Mode: head space split sampling, split ratio 10:1;Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;Air: 400mL/min, hydrogen
Gas: 30mL/min, make-up gas (N2): 25mL/min.
Test solution headspace sampling prepared by step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
Embodiment 3
1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol solution in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 0.5% internal standard stock solution
2) preparation method of standard solution: precision measures a certain amount of ethyl alcohol in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 5% ethyl alcohol standard solution.Respectively measure ethyl alcohol standard solution in right amount and a certain amount of internal standard stock solution,
It is configured to the series standard solution that internal standard concentration is 0.01%, concentration of alcohol is 0.005%-0.2%.
3) preparation method of test solution: precision weighs 0.5g sample in 100mL volumetric flask, adds a certain amount of internal standard
Stock solution, with ultrapure water constant volume, internal standard solution concentration is 0.01%, shakes up to obtain test solution.
4) determination step:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);Injector temperature: 200 DEG C;Detector: FID, temperature 220
℃;Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;Sample introduction
Mode: head space split sampling, split ratio 10:1;Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;Air: 400mL/min, hydrogen
Gas: 30mL/min, make-up gas (N2): 25mL/min.
Test solution headspace sampling prepared by step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
Embodiment 4
1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol solution in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 0.5% internal standard stock solution
2) preparation method of standard solution: precision measures a certain amount of ethyl alcohol in 100mL volumetric flask, fixed with ultrapure water
Hold, shakes up, obtain 5% ethyl alcohol standard solution.Respectively measure ethyl alcohol standard solution in right amount and a certain amount of internal standard stock solution,
It is configured to the series standard solution that internal standard concentration is 0.01%, concentration of alcohol is 0.005%-0.2%.
3) preparation method of test solution: precision weighs 1.0g sample in 100mL volumetric flask, adds a certain amount of internal standard
Stock solution, with ultrapure water constant volume, internal standard solution concentration is 0.01%, shakes up to obtain test solution.
4) determination step:
Chromatographic condition:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);Injector temperature: 200 DEG C;Detector: FID, temperature 220
℃;Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;Sample introduction
Mode: head space split sampling, split ratio 10:1;Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;Air: 400mL/min, hydrogen
Gas: 30mL/min, make-up gas (N2): 25mL/min.
Test solution headspace sampling prepared by step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
The measurement result of 1 Examples 1 to 4 of table
Project | Embodiment 1 | Embodiment 2 | Embodiment 3 | Embodiment 4 |
Ethyl alcohol percentage composition | 0.153% | 0.149% | 0.152% | 0.153% |
Conclusion: can be determined that hemostatic material sample-adding amount according to the above results, peak area ratio is in proportion in 0.25g-1.0g
Increase, test sample concentration can be detected accurately between 0.25%-1%, and the change of inner mark solution concentration, will not be influenced
Final detection result;It can be seen that the inventive method is suitable for detecting the ethyl alcohol residual of hemostatic material.
Chromatographic condition: pass through methodology validation, under chromatographic condition of the invention, each tested equal separating degree of ingredient is greater than
1.5, other compositions are noiseless to ingredient to be measured, and theoretical cam curve is higher than 10000 in terms of ethyl alcohol peak, it was demonstrated that the GC conditions
With good system suitability, suitable for the content of measurement ethyl alcohol, by the remaining control of ethyl alcohol in hemostatic material, energy
Enough quality for preferably controlling the hemostatic material.
The rate of recovery: it takes the test sample 0.1-1.0g of known Residual ethanol in 100mL volumetric flask, is added thereto respectively
100 μ L ethyl alcohol Standard Stock solutions, measure according to the method for the present invention, with measured value and test sample content difference divided by the ethyl alcohol of addition
The amount of standard items calculates the rate of recovery.Rate of recovery result illustrates that the experimental method relative error is smaller, true between 95%-105%
Reality is good.
The above described is only a preferred embodiment of the present invention, being not that the invention has other forms of limitations, appoint
What those skilled in the art changed or be modified as possibly also with the technology contents of the disclosure above equivalent variations etc.
Imitate embodiment.But without departing from the technical solutions of the present invention, according to the technical essence of the invention to above embodiments institute
Any simple modification, equivalent variations and the remodeling made, still fall within the protection scope of technical solution of the present invention.
Claims (5)
1. the detection method of Residual ethanol in a kind of hemostatic material, it is characterised in that: steps are as follows:
(1) preparation method of internal standard stock solution: taking a certain amount of normal propyl alcohol, with ultrapure water constant volume, shakes up, and is prepared into concentration and is
0.25%~2% solution, as internal standard stock solution;
(2) preparation method of standard solution: precision measures a certain amount of ethyl alcohol and is shaken up with ultrapure water constant volume, and being prepared into concentration is
5% solution, as ethyl alcohol Standard Stock solutions;Respectively measure ethyl alcohol Standard Stock solutions in right amount and a certain amount of internal standard deposit
Solution is configured to the series standard solution that internal standard concentration is identical, concentration of alcohol is 0.0025%~0.5%;
(3) preparation method of test solution: precision weighs appropriate test sample, adds a certain amount of internal standard stock solution, and use is ultrapure
Water constant volume keeps internal standard concentration identical as internal standard concentration in standard items;5~60min of extraction is shaken up, test solution is obtained;
(4) the test solution headspace sampling for preparing step (3) records chromatogram;Separately take the same method of ethanol series standard solution
Measurement obtains the residual of ethyl alcohol in hemostatic material according to internal standard method with peak area (ethyl alcohol peak area/normal propyl alcohol peak area) calculating
Amount.
2. according to claim 1 in hemostatic material Residual ethanol detection method, it is characterised in that: the step (4)
In chromatographic condition are as follows:
Chromatographic column: HP-5 (30m × 0.25mm × 0.25 μm);
Injector temperature: 200 DEG C;Detector: 220 DEG C of FID, temperature;
Column temperature: 50 DEG C of initial temperature, 2min is maintained, 80 DEG C is warming up to 5 DEG C/min, is warming up to 200 DEG C with 35 DEG C/min;
Input mode: head space split sampling, split ratio 10:1~20:1;
Ml headspace bottle temperature: 85 DEG C;Equilibration time: 30min;
Air: 400mL/min, hydrogen: 30mL/min, make-up gas (N2): 25mL/min.
3. according to claim 2 in hemostatic material Residual ethanol detection method, it is characterised in that: the step (4)
In, using head space split sampling, split ratio 10:1.
4. according to claim 1 in hemostatic material Residual ethanol detection method, it is characterised in that: the step (1)
In, the concentration of the internal standard stock solution of preparation is 0.5%.
5. according to claim 1 in hemostatic material Residual ethanol detection method, it is characterised in that: the step (2)
In, the series standard solution of preparation is internal standard concentration 0.01%, concentration of alcohol is 0.005%~0.2% serial solution.
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CN115372519A (en) * | 2022-09-21 | 2022-11-22 | 广西壮族自治区水牛研究所 | Gas chromatography detection method for ethanol in silage |
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Application publication date: 20190910 |