CN109115714A - A method of measurement hydroxyethyl starch concentration - Google Patents
A method of measurement hydroxyethyl starch concentration Download PDFInfo
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- CN109115714A CN109115714A CN201811347687.0A CN201811347687A CN109115714A CN 109115714 A CN109115714 A CN 109115714A CN 201811347687 A CN201811347687 A CN 201811347687A CN 109115714 A CN109115714 A CN 109115714A
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- China
- Prior art keywords
- distilled water
- serum
- anthrone
- room temperature
- hydroxyethyl starch
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- Pending
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- 238000000034 method Methods 0.000 title claims abstract description 25
- 229920001612 Hydroxyethyl starch Polymers 0.000 title claims abstract description 11
- 229940050526 hydroxyethylstarch Drugs 0.000 title claims abstract description 11
- 238000005259 measurement Methods 0.000 title claims description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 28
- 239000012153 distilled water Substances 0.000 claims abstract description 23
- 210000002966 serum Anatomy 0.000 claims abstract description 23
- RJGDLRCDCYRQOQ-UHFFFAOYSA-N anthrone Chemical compound C1=CC=C2C(=O)C3=CC=CC=C3CC2=C1 RJGDLRCDCYRQOQ-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000005457 ice water Substances 0.000 claims abstract description 16
- 238000002835 absorbance Methods 0.000 claims abstract description 11
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims abstract description 8
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 7
- RQPZNWPYLFFXCP-UHFFFAOYSA-L barium dihydroxide Chemical compound [OH-].[OH-].[Ba+2] RQPZNWPYLFFXCP-UHFFFAOYSA-L 0.000 claims abstract description 6
- 229910001863 barium hydroxide Inorganic materials 0.000 claims abstract description 6
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 6
- 239000000706 filtrate Substances 0.000 claims abstract description 6
- 210000002700 urine Anatomy 0.000 claims abstract description 6
- NWONKYPBYAMBJT-UHFFFAOYSA-L zinc sulfate Chemical compound [Zn+2].[O-]S([O-])(=O)=O NWONKYPBYAMBJT-UHFFFAOYSA-L 0.000 claims abstract description 6
- 229960001763 zinc sulfate Drugs 0.000 claims abstract description 6
- 229910000368 zinc sulfate Inorganic materials 0.000 claims abstract description 6
- 238000010438 heat treatment Methods 0.000 claims abstract description 5
- 238000004090 dissolution Methods 0.000 claims abstract description 3
- 210000003462 vein Anatomy 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 19
- 238000009835 boiling Methods 0.000 claims description 6
- 239000013558 reference substance Substances 0.000 claims description 6
- 210000004369 blood Anatomy 0.000 claims description 5
- 239000008280 blood Substances 0.000 claims description 5
- 230000023555 blood coagulation Effects 0.000 claims description 5
- 238000010521 absorption reaction Methods 0.000 claims description 4
- 239000000203 mixture Substances 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 3
- 238000010790 dilution Methods 0.000 claims description 3
- 239000012895 dilution Substances 0.000 claims description 3
- 238000007710 freezing Methods 0.000 claims description 3
- 230000008014 freezing Effects 0.000 claims description 3
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 claims 1
- 102000002322 Egg Proteins Human genes 0.000 claims 1
- 108010000912 Egg Proteins Proteins 0.000 claims 1
- 210000001367 artery Anatomy 0.000 claims 1
- 235000014103 egg white Nutrition 0.000 claims 1
- 210000000969 egg white Anatomy 0.000 claims 1
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 239000000686 essence Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- NNBFNNNWANBMTI-UHFFFAOYSA-M brilliant green Chemical compound OS([O-])(=O)=O.C1=CC(N(CC)CC)=CC=C1C(C=1C=CC=CC=1)=C1C=CC(=[N+](CC)CC)C=C1 NNBFNNNWANBMTI-UHFFFAOYSA-M 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 150000005846 sugar alcohols Chemical class 0.000 description 1
- 239000012085 test solution Substances 0.000 description 1
Classifications
-
- 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
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)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention discloses a kind of methods for measuring hydroxyethyl starch concentration, take anthrone concentrated sulfuric acid dissolution, dilute ten thousand rivers in Shangdong Province, and anthrone color developing agent is added and shakes up, makees blank by the same method with distilled water, cuvette colorimetric carries out recurrence calculating, obtains regression equation；Serum or urine are accurately drawn when vein hematometry, distilled water and barium hydroxide solution is added to mix, again plus solution of zinc sulfate, it is centrifuged after mixing, obtains deproteinized serum filtrate, then albumen serum is removed to mix with anthrone reagent, heating, is immediately placed in ice water, equilibrium at room temperature, make blank cuvette colorimetric by the same method with distilled water, ultraviolet light spectrophotometric instrument surveys absorbance.It is easy to operate the beneficial effects of the invention are as follows precision height.
Description
Technical field
The invention belongs to field of biotechnology, are related to a kind of method for measuring hydroxyethyl starch concentration.
Background technique
Hydroxyethyl starch is the polymer substance extracted from corn, is that one kind is widely used in severe section, fiber crops over more than 50 years
The clinical common medicine of liquor-saturated section.Hydroxyethyl starch is according to relative molecular mass and replaces the difference of grade that can be divided into many types: the 3rd
Low replacing grade on behalf of middle relative molecular mass, i.e., the 2005 HES solution (130/0.4) released, ten thousand rivers in Shangdong Province are exactly (130/
0.4), compared with HES (200/0.5), the molecular distribution of ten thousand rivers in Shangdong Province is more concentrated, and replaces grade to be reduced to 0.4 from 0.5, and replace
Mode increases to 9:1 from 5:1, replaces grade higher, and capacity enlargement effect is better, influences on hepatic and renal function smaller.
Summary of the invention
The purpose of the present invention is to provide a kind of methods for measuring hydroxyethyl starch concentration, and the beneficial effects of the invention are as follows essences
Density is high, easy to operate.
The technical scheme adopted by the invention is that following the steps below:
Step 1) takes anthrone concentrated sulfuric acid dissolution, dilutes ten thousand rivers in Shangdong Province, take not ten thousand river in Shangdong Province of same amount with distilled water supply 1ml, be placed in
Have in plug 10ml test tube, whole test tubes are put in ice water, each anthrone color developing agent that is added shakes up, then test tube is put into boiling water bath
It boiling, is immediately placed in ice water after taking-up, equilibrium at room temperature makees blank with distilled water by the same method, and cuvette colorimetric measures trap A,
Absorbance A is corresponded to reference substance solution concentration C and carries out recurrence calculating, obtains regression equation；
Step 2) venous blood is put into dry graduated centrifuge tube natural blood coagulation, centrifugation at room temperature, draws upper serum freezing
Spare, when measurement, accurately draws serum or urine, and distilled water and barium hydroxide solution is added to mix, then plus solution of zinc sulfate, mix
Centrifugation, obtains deproteinized serum filtrate, then removes albumen serum and mix with anthrone reagent after even, heats, is immediately placed in ice water
In, equilibrium at room temperature makees blank cuvette colorimetric with distilled water by the same method, and ultraviolet light spectrophotometric instrument surveys absorbance.
Further, in step 1), anthrone 1g concentrated sulfuric acid 500ml is taken to dissolve, ten thousand river in Shangdong Province concentration of dilution are 0.1mg/ml, respectively
It takes ten thousand river in Shangdong Province 0.05 0.1mg/ml, 0.1,0.2,0.3,0.4,0.6,0.8ml to supply 1ml with distilled water, has been placed in plug 10ml examination
Whole test tubes are put in ice water by Guan Zhong, and each anthrone color developing agent 4m that is added shakes up, then test tube is put into boiling water bath and boils 10min,
It is immediately placed in 5min in ice water, equilibrium at room temperature 10min after taking-up, blank, 0.5cm cuvette, 620nm are made by the same method with distilled water
Wavelength colorimetric measures trap A, corresponds to absorbance A with reference substance solution concentration C and carries out recurrence calculating, obtains regression equation.
Further, regression equation: A=0.021+9.89C, r=0.999.
Further, step 2) medium sized vein blood 3ml, is put into the dry graduated centrifuge tube of 5ml natural blood coagulation at room temperature, after 2h from
Heart 15min, absorption upper serum are frozen in spare in -20 DEG C of refrigerators.Serum or urine 0.5ml are accurately drawn when measurement, are added
Distilled water 2.5ml and barium hydroxide solution 1ml are mixed, then plus solution of zinc sulfate 1ml, be centrifuged 10min after mixing, obtain deproteinized
Then serum filtrate takes 0.1ml deproteinized serum to mix with 4ml anthrone reagent, 100 DEG C of heating 10min are immediately placed in ice water
5min, equilibrium at room temperature 10min, make blank 0.5cm cuvette with distilled water by the same method, 620nm wavelength colorimetric, and UV2100 is ultraviolet
Line spectrophotometric instrument surveys absorbance.
Specific embodiment
The present invention is described in detail With reference to embodiment.
The preparation of standard curve
Anthrone 1g concentrated sulfuric acid 500ml is taken to dissolve, ten thousand river in Shangdong Province concentration of dilution are 0.1mg/ml, take ten thousand river in Shangdong Province 0.1mg/ml respectively
0.05,0.1,0.2,0.3,0.4,0.6,0.8ml supplies 1ml with distilled water, has been placed in plug 10ml test tube, by whole test tubes
It is put in ice water, each anthrone color developing agent 4m that is added shakes up, then test tube is put into boiling water bath and boils 10min, is immediately placed in after taking-up
5min in ice water, equilibrium at room temperature 10min make blank, 0.5cm cuvette with distilled water by the same method, and 620nm wavelength colorimetric measures
Trap A corresponds to absorbance A with reference substance solution concentration C and carries out recurrence calculating, obtains regression equation.Regression equation: A=
0.021+9.89C, r=0.999.
Measuring method
Venous blood 3ml is put into the dry graduated centrifuge tube of 5ml natural blood coagulation at room temperature, is centrifuged 15min after 2h, in absorption
Layer serum is frozen in spare in -20 DEG C of refrigerators.Serum (or urine) 0.5ml is accurately drawn when measurement, add distilled water 2.5ml and
Barium hydroxide solution 1ml mix, then plus solution of zinc sulfate 1ml, be centrifuged 10min after mixing, obtain deproteinized serum filtrate, then
0.1ml deproteinized serum is taken to mix with 4ml anthrone reagent, 100 DEG C of heating 10min are immediately placed in 5min in ice water, equilibrium at room temperature
10min makees blank 0.5cm cuvette, 620nm wavelength colorimetric, UV2100 ultraviolet light spectrophotometer with distilled water by the same method
Device surveys absorbance.
Measuring method precision of the present invention is high, easy to operate.Polysaccharide component is first hydrolyzed into list under the action of sulfuric acid
Sugar, and rapid dehydration generates alditol derivative, and emerald green solution is then condensed into anthrone reagent, this solution colour is stablized, and
Reference substance and test solution have characteristic absorption at wavelength 620nm.It can be measured the lower limit of polysaccharide concentration with the method for anthrone
50 μ g/ml, the coefficient of variation 2.9%.
The above is only not to make limit in any form to the present invention to better embodiment of the invention
System, any simple modification that embodiment of above is made according to the technical essence of the invention, equivalent variations and modification,
Belong in the range of technical solution of the present invention.
Claims (4)
1. a kind of method for measuring hydroxyethyl starch concentration, it is characterised in that follow the steps below:
Step 1) takes anthrone concentrated sulfuric acid dissolution, dilutes ten thousand rivers in Shangdong Province, take not ten thousand river in Shangdong Province of same amount with distilled water supply 1ml, be placed in plug
In 10ml test tube, whole test tubes are put in ice water, each anthrone color developing agent that is added shakes up, then test tube is put into boiling water bath and is boiled,
It being immediately placed in ice water after taking-up, equilibrium at room temperature, blank is made by the same method with distilled water, cuvette colorimetric measures trap A, with
Reference substance solution concentration C corresponds to absorbance A and carries out recurrence calculating, obtains regression equation；
Step 2) venous blood is put into dry graduated centrifuge tube natural blood coagulation at room temperature, and centrifugation, absorption upper serum freezing is spare,
Accurately draw serum or urine when measurement, distilled water and barium hydroxide solution added to mix, then plus solution of zinc sulfate, after mixing from
The heart obtains deproteinized serum filtrate, then removes albumen serum and mixes with anthrone reagent, and heating is immediately placed in ice water, room temperature
Balance, makees blank cuvette colorimetric with distilled water by the same method, and ultraviolet light spectrophotometric instrument surveys absorbance.
2. according to a kind of method for measuring hydroxyethyl starch concentration described in claim 1, it is characterised in that: in the step 1),
Take anthrone 1g concentrated sulfuric acid 500ml to dissolve, ten thousand river in Shangdong Province concentration of dilution are 0.1mg/ml, take respectively ten thousand river in Shangdong Province 0.05 0.1mg/ml, 0.1,
0.2,0.3,0.4,0.6,0.8ml supplies 1ml with distilled water, has been placed in plug 10ml test tube, whole test tubes are put in ice water
In, each anthrone color developing agent 4m that is added shakes up, then test tube is put into boiling water bath and boils 10min, is immediately placed in ice water after taking-up
5min, equilibrium at room temperature 10min make blank, 0.5cm cuvette with distilled water by the same method, and 620nm wavelength colorimetric measures trap
A corresponds to absorbance A with reference substance solution concentration C and carries out recurrence calculating, obtains regression equation.
3. according to a kind of method for measuring hydroxyethyl starch concentration described in claim 1, it is characterised in that: the regression equation: A
=0.021+9.89C, r=0.999.
4. according to a kind of method for measuring hydroxyethyl starch concentration described in claim 1, it is characterised in that: quiet in the step 2)
Arteries and veins blood 3ml is put into natural blood coagulation at room temperature in the dry graduated centrifuge tube of 5ml, is centrifuged 15min after 2h, draws upper serum freezing
It is spare in -20 DEG C of refrigerators.Serum or urine 0.5ml are accurately drawn when measurement, add distilled water 2.5ml and barium hydroxide solution
1ml mix, then plus solution of zinc sulfate 1ml, be centrifuged 10min after mixing, obtain deproteinized serum filtrate, then 0.1ml is taken to remove egg
White serum is mixed with 4ml anthrone reagent, and 100 DEG C of heating 10min are immediately placed in 5min in ice water, equilibrium at room temperature 10min, to steam
Distilled water makees blank 0.5cm cuvette, 620nm wavelength colorimetric by the same method, and UV2100 ultraviolet light spectrophotometric instrument surveys absorbance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811347687.0A CN109115714A (en) | 2018-11-13 | 2018-11-13 | A method of measurement hydroxyethyl starch concentration |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811347687.0A CN109115714A (en) | 2018-11-13 | 2018-11-13 | A method of measurement hydroxyethyl starch concentration |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109115714A true CN109115714A (en) | 2019-01-01 |
Family
ID=64853993
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811347687.0A Pending CN109115714A (en) | 2018-11-13 | 2018-11-13 | A method of measurement hydroxyethyl starch concentration |
Country Status (1)
| Country | Link |
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| CN (1) | CN109115714A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101644713A (en) * | 2009-07-10 | 2010-02-10 | 杨维明 | Application of hydroxyethyl starch within molecular weight range of 60,000-80,000 |
| CN103175794A (en) * | 2011-12-20 | 2013-06-26 | 四川科伦药物研究有限公司 | Method for measuring content of sodium element in hydroxyethyl starch injection preparation |
| EP2443447B1 (en) * | 2009-06-19 | 2015-08-12 | B. Braun Melsungen AG | Use of an oxygen indicator for parenteral and enteral modes of administration |
-
2018
- 2018-11-13 CN CN201811347687.0A patent/CN109115714A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2443447B1 (en) * | 2009-06-19 | 2015-08-12 | B. Braun Melsungen AG | Use of an oxygen indicator for parenteral and enteral modes of administration |
| CN101644713A (en) * | 2009-07-10 | 2010-02-10 | 杨维明 | Application of hydroxyethyl starch within molecular weight range of 60,000-80,000 |
| CN103175794A (en) * | 2011-12-20 | 2013-06-26 | 四川科伦药物研究有限公司 | Method for measuring content of sodium element in hydroxyethyl starch injection preparation |
Non-Patent Citations (4)
| Title |
|---|
| WILKES, NJ: "Hydroxyethyl starch in balanced electrolyte solution (Hextend((R)))-pharmacokinetic and pharmacodynamic profiles in healthy volunteers", 《ANESTHESIA AND ANALGESIA》 * |
| 曾媛: "6 %羟乙基淀粉200/0.5 的药代动力学研究", 《临床麻醉学杂志》 * |
| 熊方武: "《中国临床药物大辞典 化学药卷 下》", 31 August 2018, 中国医药科技出版社 * |
| 王燕萍: "血浆中辅酶Q10和羟乙基淀粉定量分析方法研究和应用", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
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Application publication date: 20190101 |
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