CN102759518A - Resonance light scattering detection method for sodium heparin - Google Patents
Resonance light scattering detection method for sodium heparin Download PDFInfo
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- CN102759518A CN102759518A CN2012102671340A CN201210267134A CN102759518A CN 102759518 A CN102759518 A CN 102759518A CN 2012102671340 A CN2012102671340 A CN 2012102671340A CN 201210267134 A CN201210267134 A CN 201210267134A CN 102759518 A CN102759518 A CN 102759518A
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Abstract
The invention discloses a resonance light scattering detection method for sodium heparin, wherein under an alkaline condition with the NaOH concentration being 0.042mol/L, Co(II) and a cobalt reagent are bonded to produce a complex with positive charges, the sodium heparin carries negative charges and is electrostatically bonded with the complex to form a ternary complex, so that the intensity of resonance light scattering is enhanced obviously and has the maximal scattering peak located at 550nm, the scattering intensity is in good linear relation with the concentration of the sodium heparin in a range of 0.05-1.60 mu g/mL, and the detection limit (3 sigma) is 1.3ng/mL. The method has the advantages of simple required detection equipment, readily available reagent, simpleness for operation, short detection time, wider linear range and higher sensitivity. The invention enriches the detection methods of the sodium heparin and is favorable for meeting detection requirements under different conditions.
Description
Technical field
The invention belongs to analysis technical field, relate to a kind of Pharmaceutical Analysis method.
Background technology
Cardiovascular and cerebrovascular disease is human No.1 disease killer.The Along with people's living standard improves overnutrition, global environment deterioration, rhythm of life quickening, the aging population aggravation that brings, and the M & M of global cardiovascular and cerebrovascular disease increases just year by year.Liquaemin is the maximum anticoagulation medicine of at present the most effective and clinical in the world consumption, is mainly used in cardiovascular and cerebrovascular disease and hemodialysis, and is unique effective specific medicament in hemodialysis.Clinical practice and research show that liquaemin also has other multiple biologically active and clinical applications except that having blood coagulation resisting function, comprise effects such as reducing blood lipid, anti-middle film smooth muscle cell proliferation, promotion fibrinolysis.With the liquaemin is that the low molecular sodium heparin that raw material further is processed into then has clinical medicine purposes more widely, has become the choice drug of acute phlebothrombosis of treatment and acute coronary artery syndrome diseases such as (angina pectoris, miocardial infarctions).The international market is very powerful to the demand of liquaemin bulk drug in recent years, and the Lipo-Hepin consumption is steadily increasing, and the rapid dilatation in the market of low molecular sodium heparin also keeps rapid growth trend.
The molecular structure of liquaemin is complicated, in a short time can't artificial chemosynthesis, and have only the liquaemin that in pig or Roll mucous membrane, extracts to can be used in clinical treatment at present.Because it is clinical that the mode of the many at present employing drug administration by injection of liquaemin is used for, thereby require the liquaemin bulk drug must possess very high purity, can guarantee the drug safety of preparation.In addition, liquaemin can cause hematostaxis when dosage is excessive clinically, like mucosal bleeding, hemarthrosis, wound bleeding etc.Therefore, the content of accurately measuring liquaemin has very important significance.
The assay method of liquaemin has biological method and chemical method two big classes at present.The assay method that existing pharmacopeia is recorded is the heparin bioassay method, is to prolong the effect of setting time of fresh rabbit blood or rabbit, porcine blood plasma through relatively heparin standard items and test sample, measures tiring of test sample.Chemical method mainly contains high performance liquid chromatography, capillary electrophoresis, AAS, fluorescence method, light scattering method, electrochemical methods etc.For example, Liu Shaopu etc. are probe with alkaline diphenyl naphthyl methane dye and alkaline kiton colors, adopt AAS or light scattering method that liquaemin is measured; Sun Wei etc. are luminosity reagent with the dimethyl diaminophenazine chloride, in the B-R of pH3.0 buffer solution, utilize dimethyl diaminophenazine chloride and liquaemin to mutually combine and cause the phenomenon of solution decolourization, adopt the spectrophotometry liquaemin; Guo Xiaoqun etc. make toner with the methyl red dyestuff, near neutral B-R buffer solution, utilize liquaemin and methyl red dyestuff to form the phenomenon that the resonance Rayleigh intensity significantly strengthens behind the ionic associate, adopt resonance rayleigh light scattering method mensuration liquaemin.Though existing at present many bibliographical informations the assay method of multiple liquaemin; But from further improving accuracy in detection, sensitivity and degree of accuracy; Simplify the operation; Enrich detection method to satisfy the purposes such as detection demand under the different condition, still be necessary to continue to develop more, more advanced liquaemin assay methods.
Summary of the invention
In view of this; The object of the present invention is to provide a kind of resonant light scattering (RLS) detection method of liquaemin; Equipment is simple, reagent is easy to get, easy operating, detection time weak point, range of linearity broad, sensitivity is higher; To enrich the detection method of liquaemin, satisfy the detection demand under the different condition.
For achieving the above object, the present invention adopts following technical scheme:
The resonant light scattering detection method of liquaemin may further comprise the steps:
A. the making of typical curve
A1. in reaction vessel, add the WS of Co (II) and the ethanol solution of cobalt reagent, mixing adds the WS of NaOH again, and mixing adds the WS of liquaemin again, and dilute with water is processed and contained 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5Mol/L cobalt reagent, 0.042 mol/L NaOH and liquaemin concentration in gradient series standard solution in 0.05 ~ 1.60 μ g/mL scope;
A2. the blank solution that does not add liquaemin according to the said method preparation of step a1;
A3. get the series standard solution of step a1 preparation; With fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out synchronous scanning under the condition of emission wavelength, slit 10 nm; Obtain resonant light scattering spectrum, measure the scattering strength at 550nm place, be designated as
IOther gets the blank solution of step a2 preparation, and the scattering strength with method mensuration 550nm place is designated as
I 0According to formula △
I 550=
I-
I 0Calculate △
I 550, make △
I 550Typical curve to liquaemin concentration;
B. sample determination
B1. in reaction vessel, add the WS of Co (II) and the ethanol solution of cobalt reagent, mixing adds the WS of NaOH again, and mixing adds the WS of the sample of unknown liquaemin content again, and dilute with water is processed and contained 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5The solution to be measured of mol/L cobalt reagent, 0.042 mol/L NaOH and unknown liquaemin concentration;
B2. the blank solution that does not add liquaemin according to the said method preparation of step b1;
B3. get the solution to be measured of step b1 preparation; With fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out synchronous scanning under the condition of emission wavelength, slit 10 nm; Obtain resonant light scattering spectrum, measure the scattering strength at 550nm place, be designated as
IOther gets the blank solution of step b2 preparation, and the scattering strength with method mensuration 550nm place is designated as
I 0According to formula △
I 550=
I-
I 0Calculate △
I 550, the △ that obtains according to step a3 again
I 550To the typical curve of liquaemin concentration, calculate the content of liquaemin in the sample.
Further, said step a1 adds 1.0 mL 2.0 * 10 in reaction vessel
-4Mol/L CoCl
2The WS and 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent ethanol solution, mixing adds 1.4 mL, the 0.3 mol/L NaOH WS again, and mixing adds the 10 μ g/mL liquaemin WS again, is diluted with water to 10 mL, processes and contains 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5Mol/L cobalt reagent, 0.042 mol/L NaOH and liquaemin concentration are respectively the series standard solution of 0.05,0.1,0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6 μ g/mL;
Said step b1 adds 1.0 mL 2.0 * 10 in reaction vessel
-4Mol/L CoCl
2The WS and 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent ethanol solution, mixing adds 1.4 mL, the 0.3 mol/L NaOH WS again, and mixing adds the sample aqueous solution of unknown liquaemin content again, is diluted with water to 10 mL, processes and contains 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5The solution to be measured of mol/L cobalt reagent, 0.042 mol/L NaOH and unknown liquaemin concentration.
Beneficial effect of the present invention is: the present invention has set up a kind of method of utilizing resonant light scattering technology quantitative measurement liquaemin; In NaOH concentration is under the alkali condition of 0.042 mol/L; Co (II) combines the complex of generation positively charged with cobalt reagent; Liquaemin is electronegative, and the two combines to form ternary complex through static, and resonant light scattering intensity is enlarged markedly; The maximum scattering peak position is in 550 nm; Scattering strength and liquaemin concentration are good linear relationship in 0.05-1.60 μ g/mL scope, detection limit (3 σ) is 1.3 ng/mL, and required checkout equipment is simple, reagent is easy to get, easy operating, detection time weak point, range of linearity broad, sensitivity is higher.The present invention has enriched the detection method of liquaemin, is beneficial to the detection demand that satisfies under the different condition.
Description of drawings
In order to make the object of the invention, technical scheme and advantage clearer, will combine accompanying drawing that the present invention is further described below, wherein:
Fig. 1 is the interactional resonant light scattering spectrum of Co (II), cobalt reagent and liquaemin, and wherein numbering 0 is the resonant light scattering spectrum of 1.2 μ g/mL heparin sodium aquas, and numbering 1 to 7 is 2.0 * 10
-5Mol/L Co (II) and 4.0 * 10
-5The mol/L cobalt reagent is in the presence of 0.042 mol/L NaOH, respectively with the interactional resonant light scattering spectrum of 0,0.2,0.4,0.6,0.8,1.0,1.2 μ g/mL liquaemins.
Fig. 2 is that resonant light scattering intensity is with NaOH change in concentration figure.
Fig. 3 is the mol ratio variation diagram of resonant light scattering intensity with Co (II) and cobalt reagent.
Fig. 4 is the typical curve of resonant light scattering intensity to liquaemin concentration.
Embodiment
Below will carry out detailed description through concrete experiment to the present invention with reference to accompanying drawing.
The liquaemin solid that uses in the experiment is Shanghai chemical reagents corporation of Chinese Medicine group product (160 IU/mg), and its solution with water is prepared and got; Co (II) solution is CoCl
2The WS; Cobalt reagent is 4-(5-chloro-2-pyridylazo)-1, and the 3-diaminobenzene is abbreviated as 5-Cl-PADAB, and its solution gets with the absolute ethyl alcohol preparation.Resonant light scattering spectrum that relates in the experiment and intensity (
I) record by F-4500 fluorospectrophotometer (Hitachi, Ltd).
One, the interactional resonant light scattering spectral signature of Co (II), cobalt reagent and liquaemin
In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent solution and 1.4 mL, 0.3 mol/L NaOH solution; Whenever add all abundant mixing of a kind of reagent; Add an amount of 10 μ g/mL liquaemin standard solution (final concentration is respectively 0.2,0.4,0.6,0.8,1.0,1.2 μ g/mL) again; Last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals emission wavelength (△
λ=0 nm), carry out synchronous scanning under the condition of slit 10 nm and obtain resonant light scattering spectrum.Simultaneously according to the method described above but do not add liquaemin preparation blank, measure with method.
The result shows that Co (II) and cobalt reagent act on the complex that forms a kind of positively charged under alkali condition, because liquaemin is electronegative, it can form ternary complex through the complex-bound of electrostatic interaction and above-mentioned positively charged, thereby causes scattered light to strengthen.As shown in Figure 1; The scattering strength of independent liquaemin or Co (II)-cobalt reagent complex is all very low, and after the two reaction formed ternary complex, scattering strength significantly strengthened; And at wavelength 280nm and two characteristic scattering peaks of 550nm place appearance; Wherein the scattering at 550nm place strengthens more obviously, and scattering strength increases along with the increase of liquaemin concentration (0.2-1.2 μ g/mL), demonstrates certain linear relationship.
Two, testing conditions optimization
1, NaOH concentration is optimized
In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent solution and an amount of 0.3 mol/L NaOH solution (final concentration is respectively 0,0.006,0.012,0.018,0.024,0.030,0.036,0.042,0.048,0.054 mol/L); Whenever add all abundant mixing of a kind of reagent; Add 1mL 10 μ g/mL liquaemin standard solution again; Last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out under the condition of emission wavelength, slit 10 nm synchronous scanning and obtains resonant light scattering spectrum, measures the scattering strength at 550nm place.Simultaneously according to the method described above but do not add liquaemin preparation blank solution, measure with method.Draw scattering strength with NaOH change in concentration figure.
The result is as shown in Figure 2, under the neutrallty condition that does not add NaOH, adds behind the liquaemin almost no change of scattering strength; Along with the increase that adds NaOH concentration, add liquaemin after scattering strength also increase gradually, when NaOH concentration reached 0.042 mol/L, it is maximum that scattering strength reaches, and weakens gradually again afterwards.Analyzing its reason, possibly be when NaOH concentration is lower than 0.042 mol/L, combines with Co (II)-cobalt reagent complex static owing to the liquaemin deionization is unfavorable for it; When NaOH concentration increased gradually, liquaemin ionization and be with more negative charge gradually caused stronger scattered signal with positively charged complex-bound easily.Therefore, NaOH concentration is preferably 0.042 mol/L.
2, the mol ratio optimization of Co (II) and cobalt reagent
In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, an amount of 5.0 * 10
-4(final concentration is respectively 1.0,2.0,3.0,4.0,5.0,6.0,7.0 * 10 to mol/L cobalt reagent solution
-5Mol/L) and 1.4 mL, 0.3 mol/L NaOH solution, whenever add all abundant mixings of a kind of reagent, add 1mL 10 μ g/mL liquaemin standard solution again, last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out under the condition of emission wavelength, slit 10 nm synchronous scanning and obtains resonant light scattering spectrum, measures the scattering strength at 550nm place.Simultaneously according to the method described above but do not add liquaemin preparation blank solution, measure with method.Draw the mol ratio variation diagram of scattering strength with Co (II) and cobalt reagent.
The result is as shown in Figure 3, keeps Co (II) concentration constant, increases the concentration of cobalt reagent gradually, and scattering strength appears increases the trend that afterwards reduces earlier, and when the mol ratio of Co (II) and cobalt reagent was 1:2, scattering strength was maximum.Analyze its reason; Possibly be because a part Co (II) can form complex with two molecule cobalt reagent coordinations; When the two mol ratio is 1:2, form the bigger saturated complex of molecular structure just, combine with liquaemin static again; Gained ternary complex particle volume is bigger, thereby scattered signal is strengthened.Therefore, the mol ratio of Co (II) and cobalt reagent is preferably 1:2.
3, the influence of coexisting substances
In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent solution and 1.4 mL, 0.3 mol/L NaOH solution; Whenever add all abundant mixing of a kind of reagent; Add 1mL 10 μ g/mL liquaemin standard solution and an amount of coexisting substances (common metallic ion, carbohydrate, amino acid or albumen) solution again; Last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out under the condition of emission wavelength, slit 10 nm synchronous scanning and obtains resonant light scattering spectrum, measures the scattering strength at 550nm place.Simultaneously according to the method described above but do not add liquaemin preparation blank solution, measure with method.Calculate the variation percent (△ that adds coexisting substances front and back RLS intensity
I RLS%).
The result is as shown in table 1, in above-mentioned detection architecture, and K
+, Na
+, Ba
2+Isoionic allowance is higher relatively, and Ag
+, Cu
2+, Pb
2+, Mg
2+, Al
3+Isoionic allowance is relatively low, and this is that these ions form precipitation of hydroxide and the parasitic resonance light scattering signal easily because the mensuration of liquaemin is carried out under alkali condition; In addition, common carbohydrate and amino acid whose allowance are higher relatively, and the allowance of albumen is relatively low; This is because under alkali condition; The common isoelectric point of pH value greater than albumen, and make the protein band negative charge, and the structure relative complex of albumen; It also can be with positively charged after Co (II)-cobalt reagent complex generation static combines, thereby also can form the mensuration that the bigger compound of structure disturbs liquaemin.But generally speaking, the relatively low concentration of interfering material in actual sample (like heparin sodium injection) of above-mentioned allowance all is lower than concentration shown in the table 1 usually, therefore can not disturb the mensuration of liquaemin.
The influence of table 1 coexisting substances
Three, the range of linearity and detection limit
In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent solution and 1.4 mL, 0.3 mol/L NaOH solution; Whenever add all abundant mixing of a kind of reagent; Add an amount of 10 μ g/mL liquaemin standard solution (final concentration is respectively 0.05,0.1,0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6 μ g/mL) again; Last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out under the condition of emission wavelength, slit 10 nm synchronous scanning and obtains resonant light scattering spectrum, measures the scattering strength at 550nm place, be designated as
ISimultaneously according to the method described above but do not add liquaemin preparation blank solution, measure the scattering strength at 550nm place, be designated as with method
I 0Calculate △
I 550=
I-
I 0, make △
I 550Typical curve to liquaemin concentration.
The result is as shown in Figure 4, is optimizing under the testing conditions, and liquaemin concentration is in 0.05-1.60 μ g/mL scope and △
I 550Be good linear relationship, linear equation is △
I=-306.1+2249.1
c, detection limit (3 σ) is 1.3 ng/mL, related coefficient is 0.9950.
Four, actual sample is measured
Get the heparin sodium injection that two commercially available different manufacturers (biochemical incorporated company of Tianjin biochemical-pharmaceutical factory and Jiangsu ten thousand nations) are produced, adopt the inventive method to detect the content of liquaemin.In 10.0 mL test tubes, add 1.0 mL 2.0 * 10 successively
-4Mol/L Co (II) solution, 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent and 1.4 mL, 0.3 mol/L NaOH solution; Whenever add all abundant mixing of a kind of reagent; Per sample liquaemin labelled amount adds an amount of sample (making the liquaemin final concentration in 0.05-1.60 μ g/mL scope) again, and last water constant volume is to 10mL, abundant mixing.With the F-4500 fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out under the condition of emission wavelength, slit 10 nm synchronous scanning and obtains resonant light scattering spectrum, measures the scattering strength at 550nm place, be designated as
ISimultaneously according to the method described above but do not add liquaemin preparation blank solution, measure the scattering strength at 550nm place, be designated as with method
I 0Calculate △
I 550=
I-
I 0, the △ that obtains according to the front again
I 550To the typical curve of liquaemin concentration, calculate the liquaemin content of sample.
The result is as shown in table 2; Adopt the inventive method to measure the content of two different manufacturers heparin sodium injections (labelled amount is 12500 IU/2mL); Average recovery rate is respectively 104.0% (RSD 2.6%) and 101.0% (RSD 2.1%), explains that the inventive method is a kind of accurate, reliable liquaemin method for quantitatively determining.
The assay result of table 2 heparin sodium injection (
n=5)
Explanation is at last; Above embodiment is only unrestricted in order to technical scheme of the present invention to be described; Although through invention has been described with reference to the preferred embodiments of the present invention; But those of ordinary skill in the art should be appreciated that and can make various changes to it in form with on the details, and the spirit and scope of the present invention that do not depart from appended claims and limited.
Claims (2)
1. the resonant light scattering detection method of liquaemin is characterized in that, may further comprise the steps:
A. the making of typical curve
A1. in reaction vessel, add the WS of Co (II) and the ethanol solution of cobalt reagent, mixing adds the WS of NaOH again, and mixing adds the WS of liquaemin again, and dilute with water is processed and contained 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5Mol/L cobalt reagent, 0.042 mol/L NaOH and liquaemin concentration in gradient series standard solution in 0.05 ~ 1.60 μ g/mL scope;
A2. the blank solution that does not add liquaemin according to the said method preparation of step a1;
A3. get the series standard solution of step a1 preparation; With fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out synchronous scanning under the condition of emission wavelength, slit 10 nm; Obtain resonant light scattering spectrum, measure the scattering strength at 550nm place, be designated as
IOther gets the blank solution of step a2 preparation, and the scattering strength with method mensuration 550nm place is designated as
I 0According to formula △
I 550=
I-
I 0Calculate △
I 550, make △
I 550Typical curve to liquaemin concentration;
B. sample determination
B1. in reaction vessel, add the WS of Co (II) and the ethanol solution of cobalt reagent, mixing adds the WS of NaOH again, and mixing adds the WS of the sample of unknown liquaemin content again, and dilute with water is processed and contained 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5The solution to be measured of mol/L cobalt reagent, 0.042 mol/L NaOH and unknown liquaemin concentration;
B2. the blank solution that does not add liquaemin according to the said method preparation of step b1;
B3. get the solution to be measured of step b1 preparation; With fluorospectrophotometer in 220-700 nm scope, excitation wavelength equals to carry out synchronous scanning under the condition of emission wavelength, slit 10 nm; Obtain resonant light scattering spectrum, measure the scattering strength at 550nm place, be designated as
IOther gets the blank solution of step b2 preparation, and the scattering strength with method mensuration 550nm place is designated as
I 0According to formula △
I 550=
I-
I 0Calculate △
I 550, the △ that obtains according to step a3 again
I 550To the typical curve of liquaemin concentration, calculate the content of liquaemin in the sample.
2. the resonant light scattering detection method of liquaemin according to claim 1 is characterized in that, said step a1 adds 1.0 mL 2.0 * 10 in reaction vessel
-4Mol/L CoCl
2The WS and 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent ethanol solution, mixing adds 1.4 mL, the 0.3 mol/L NaOH WS again, and mixing adds the 10 μ g/mL liquaemin WS again, is diluted with water to 10 mL, processes and contains 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5Mol/L cobalt reagent, 0.042 mol/L NaOH and liquaemin concentration are respectively the series standard solution of 0.05,0.1,0.2,0.4,0.6,0.8,1.0,1.2,1.4,1.6 μ g/mL;
Said step b1 adds 1.0 mL 2.0 * 10 in reaction vessel
-4Mol/L CoCl
2The WS and 0.8 mL 5.0 * 10
-4Mol/L cobalt reagent ethanol solution, mixing adds 1.4 mL, the 0.3 mol/L NaOH WS again, and mixing adds the sample aqueous solution of unknown liquaemin content again, is diluted with water to 10 mL, processes and contains 2.0 * 10
-5Mol/L Co (II), 4.0 * 10
-5The solution to be measured of mol/L cobalt reagent, 0.042 mol/L NaOH and unknown liquaemin concentration.
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| CN104076014A (en) * | 2014-06-30 | 2014-10-01 | 安徽师范大学 | Fluorescence sensor, as well as preparation method and application of sensor and method for detecting Heparin molecule |
| CN104655602A (en) * | 2015-02-05 | 2015-05-27 | 嘉兴学院 | Hydrophobic ionic liquid (MBPy) Tf2N fluorescence detection method |
| CN111684277A (en) * | 2018-03-01 | 2020-09-18 | 埃科莱布美国股份有限公司 | Method for measuring benzimidazole compounds in water |
| CN113008852A (en) * | 2021-02-23 | 2021-06-22 | 集美大学 | Detection method of vibrio phage titer |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104076014A (en) * | 2014-06-30 | 2014-10-01 | 安徽师范大学 | Fluorescence sensor, as well as preparation method and application of sensor and method for detecting Heparin molecule |
| CN104655602A (en) * | 2015-02-05 | 2015-05-27 | 嘉兴学院 | Hydrophobic ionic liquid (MBPy) Tf2N fluorescence detection method |
| CN104655602B (en) * | 2015-02-05 | 2017-06-27 | 嘉兴学院 | A kind of hydrophobic ionic liquid [MBPy] Tf2The fluorescence detection method of N |
| CN111684277A (en) * | 2018-03-01 | 2020-09-18 | 埃科莱布美国股份有限公司 | Method for measuring benzimidazole compounds in water |
| CN111684277B (en) * | 2018-03-01 | 2023-10-20 | 埃科莱布美国股份有限公司 | Method for measuring benzimidazole compounds in water |
| CN113008852A (en) * | 2021-02-23 | 2021-06-22 | 集美大学 | Detection method of vibrio phage titer |
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