CN103149170B - Solution concentration of nadroparin calcium determined by phenanthroline-zinc sulfate ultraviolet spectroscopy - Google Patents
Solution concentration of nadroparin calcium determined by phenanthroline-zinc sulfate ultraviolet spectroscopy Download PDFInfo
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Abstract
The invention discloses solution concentration of nadroparin calcium determined by phenanthroline-zinc sulfate ultraviolet spectroscopy. Under the environment of buffered solution, Ultraviolet absorbance degrees of mixed solution which includes Zn2+ions, phenanthroline and the nadroparin calcium are determined, and then the concentration of the nadroparin calcium is determined by quantity. According to the solution concentration of the nadroparin calcium determined by the phenanthroline-zinc sulfate ultraviolet spectroscopy, a phenanthroline-zinc sulfate ultraviolet spectroscopy test system is applied, quick quantitative determination of the solution concentration of the nadroparin calcium can be achieved. The linear range of an analytical method of the solution concentration of the nadroparin calcium determined by the phenanthroline-zinc sulfate ultraviolet spectroscopy is 1.184%-16.33%, meanwhile, the analytical method of the solution concentration of the nadroparin calcium determined by the phenanthroline-zinc sulfate ultraviolet spectroscopy has good anti-jamming capability and can be applied in the practical production process of nadroparin calcium medicines and capable of carrying out concentration determination to intermediate product solution or finished product solution.
Description
Technical field
The invention belongs to the production field of nadroparin calcium medicine, particularly relate to a kind of method applying Phen-zinc sulfate ultraviolet spectroscopy quantitative determination nadroparin calcium solution concentration.
Background technology
The mucopolysaccharide sulfuric ester that heparin (Heparin) is alternately made up of GLUCOSAMINE, L-iduronic acid, N-acetyl-glucosamine and glucuronic acid, molecular weight is from 5 to 30kDa, and wherein sulfate radical accounts for 40%.Usually, heparin class medicine derives from pig, Roll mucous membrane and lungs at first.Medically heparin, mainly as anticoagulant, is formally applied to clinical treatment in nineteen fifty-three current, is considered to the most effectively and anticoagulation medicine that quantity is maximum.Use unfractionated heparin at first, i.e. Unfractionated Heparin.To last century Mo, by the method such as chemical cracking or biological enzymolysis, unfractionated heparin is prepared into the heparin derivatives that mean molecular weight is distributed as 3500 ~ 6500, i.e. LMWHs (Low Molecular Weight Heparin), its drug effect obviously obtains enhancing, becomes the novel heparin class medicine be widely used.According to statistics, the current whole world about has the heparin bulk drug of 1/3rd to be used for making standard heparin preparation of sodium, all the other about 2/3rds as raw material for the production of Low-molecular-weight Heparins Calcium (low molecular sodium heparin) bulk drug.Further data show, in 2008, global heparin bulk drug sales volume was about 800,000,000 dollars, and global heparin class sales amount of medicine is then close to 6,000,000,000 dollars simultaneously; Estimate 2012, the sales volume of global heparin class medicine will more than 9,000,000,000 dollars.The general flow sheet of LMWHs class medicine as shown in Figure 1.
Wherein, nadroparin calcium (Nadroparin Calcium) is the LMWHs medicine adopting nitrous acid cleavage method to prepare, and its principal character comprises: average molecular weight range is 3600-5000Da; The reduction end of molecular structure is 6-O-sulfuric acid-2,5-dehydrations-PEARLITOL 25C, and non-reducing end is 2-O-sulfuric acid-α-L-iduronic acid; Anti-Xa tire be 95 ~ 130IU/mg, AXa/A II a numerical value be 2.5 ~ 4.0.The nadroparin calcium medicine that our company produces reaches corresponding quality index, and its simple technological process of production as shown in Figure 2.In actual production process, need to carry out the techniques such as ultrafiltration, alcohol precipitation and drying, we have found the nadroparin calcium solution obtained after ultrafiltration technology, need to carry out rapidly Concentration Testing accurately, and according to gained test result, suitable concentration regulation and control are carried out to it, so extremely be conducive to the control of subsequent technique, especially alcohol precipitation process.But in the production run of reality, the nadroparin calcium solution obtained after ultrafiltration technology is difficult to by analytical approachs such as solution weight method or high performance liquid chromatographies, completes this Detection task rapidly.
On the other hand, the initial source of heparin class medicine is mainly all the animal materials such as pig intestinal mucosa both at home and abroad at present, there is geographic difference, in nitrogen content, sulfur content and glycosyl sequence etc., has larger variation range.Simultaneously, nadroparin calcium medicine is in fact also one and defines molecular weight ranges and have the macromolecule mixture of other main character, therefore the detection method for nadroparin calcium must possess stronger antijamming capability, otherwise in the testing process of actual sample, can produce larger analytical error, gained test result can not be applied in technique to be controlled and instructs subsequent technique.
Summary of the invention
The object of the invention is to for the above-mentioned deficiency in existing nadroparin calcium actual production, provide a kind of Phen-zinc sulfate ultraviolet spectroscopy to measure the solution concentration of nadroparin calcium.
In order to achieve the above object, present invention employs following technical scheme:
Phen-zinc sulfate ultraviolet spectroscopy measures the solution concentration of nadroparin calcium, under the environment of buffer solution, by measuring Zn
2+the ultraviolet absorptivity of ion, Phen and nadroparin calcium three mixed solution carrys out the concentration of quantitative measurement nadroparin calcium.
Particularly, under the environment of buffer solution, by Zn
2+solion, Phen solution and nadroparin calcium solution mix, Zn in mixed solution
2+the concentration range of ion is at 0.05 ~ 0.3mM, and the amount of substance of Phen is Zn
2+more than 2 times of the amount of substance of ion, measure the ultraviolet absorptivity of mixed solution;
During Criterion working curve, add the nadroparin calcium solution for concentration known of mixed solution, the quality of nadroparin calcium is relative to Zn
2+the amount of substance of ion is 11.84g/mmol ~ 163.3g/mmol;
After Criterion working curve, add the nadroparin calcium solution for unknown concentration of mixed solution, calculate the concentration of nadroparin calcium solution according to the absorbance of mensuration and the standard working curve of foundation, when the quality of the nadroparin calcium added is relative to Zn
2+the amount of substance of ion is not when the scope of 11.84g/mmol ~ 163.3g/mmol, change by method that is concentrated or dilution the concentration adding nadroparin calcium solution in mixed solution when not changing and adding volume, measure its absorbance and utilize standard working curve to calculate the concentration of nadroparin calcium solution, making the quality of nadroparin calcium relative to Zn
2+the amount of substance of ion is in the scope of 11.84g/mmol ~ 163.3g/mmol;
It is pointed out that the concentration of nadroparin calcium refers to the quality percent by volume of nadroparin calcium.
Particularly, the solvent of mixed solution is at least one in water, ethanol, methyl alcohol, isopropyl alcohol, n-propanol and normal butyl alcohol.
Particularly, buffer solution comprises the aqueous solution of at least one material in sodium acetate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium carbonate, sodium bicarbonate and niter cake.
Preferably, buffer solution is sodium acetate solution, and the concentration of sodium acetate in mixed solution is 0.05 ~ 0.6M.
Preferably, the concentration of sodium acetate in mixed solution is 0.3M.
Particularly, Zn
2+ion from zinc sulfate, Zn
2+the concentration of ion is 0.2mM.
Preferably, the amount of substance of Phen is Zn
2+4 times of the amount of substance of ion.
Preferably, when measuring the absorbance of mixed solution, ultraviolet wavelength is 302 ± 1nm.
Preferably, during preparation mixed solution, zinc sulfate solution is 1.0mL, 2.0mM, the ethanolic solution of Phen is 1.0mL, 8.0mM, aqueous sodium acetate solution is 3.0mL, 1.0M, fully after mixing, adds 2.0mL nadroparin calcium solution and 3.0mL water for injection, abundant mixing also, after leaving standstill, measures the absorbance of mixed solution at 302 ± 1nm.
The invention discloses the solution concentration that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium, this assay method application Phen-zinc sulfate ultraviolet spectrum test system, achieve the Quantitative detection to nadroparin calcium solution concentration, the range of linearity of its analytical approach is 1.184% ~ 16.33%; Meanwhile, the method has good antijamming capability to calcium ion, sodion and sulfate ion, can be applied in the actual production process of nadroparin calcium medicine, carries out Concentration Testing to middle product solution or final mean annual increment solution.
Accompanying drawing explanation
Fig. 1 is LMWHs class medicament universal flow sheet;
Fig. 2 is the production technological process of nadroparin calcium medicine;
Fig. 3 is the ultraviolet spectrum detection figure of different batches nadroparin calcium sample in Phen-solution of zinc sulfate;
Fig. 4 is variable concentrations calcium ion affects detection figure to what detect solution;
Fig. 5 is different sodium acetate solution consumption affects detection figure to what detect solution;
Fig. 6 is different Phen solution usage affects detection figure to what detect solution;
Fig. 7 is different sodium nitrite solution consumption affects detection figure to what detect solution;
Fig. 8 is the detection figure of variable concentrations nadroparin calcium sample in Phen-solution of zinc sulfate.
Embodiment
Below in conjunction with specific embodiment, technical scheme of the present invention is made further clear and complete instructions.
First it should be noted that, the liquaemin sample that in the present invention, production technology source is used is from Hebei Changshan Biochemical Pharmaceutical Co., Ltd.; Nadroparin calcium sample, from Shenzhen Sciprogen Biology Medicine Co., Ltd.Meanwhile, the Primary Chemical used in the present invention comprises: anhydrous sodium acetate (CH
3cOONa, analyzes pure, CAS127-09-3); Phen (C
12h
8n
2h
2o, analyzes pure, when CAS5144-89-8 uses, is mixed with ethanolic solution); White vitriol (ZnSO
47H
2o, analyzes pure, CAS7446-20-0); Lime chloride (CaCl
2, pharmaceutical grade, CAS10043-52-4).In embodiment, the concentration of nadroparin calcium solution refers to the quality percent by volume of nadroparin calcium.
Embodiment
Embodiment discloses the solution concentration that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium, first by experiment the impact of the applicability of Phen-zinc sulfate system, anti-interference on calcium ion, the impact of sodium acetate solution consumption, the impact of Phen consumption and sodium nitrite solution consumption is checked in one to experiment five successively, so that the applicability of this detection system, alterable scope and Cleaning Principle to be described; Then six standard working curves setting up absorbance and nadroparin calcium solution concentration by experiment, the reliability of the method and this standard working curve is verified in experiment seven by experiment.
The applicability of experiment one, Phen-zinc sulfate system
Get 6 15mL centrifuge tubes, wherein 5 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate, 1.0mL8.0mM Phen solution and 3.0mL1.0M sodium acetate solution respectively, fully mix.Then, toward in sample test system, the nadroparin calcium sample solution of the different lot number of 2.0mL is added respectively, and 3.0mL water for injection, shake up; Toward in blank system, add 5.0mL water for injection, fully mix.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 5 sample test systems respectively.Gained test result as shown in Figure 3.
Can find from Fig. 3, in Phen-zinc sulfate system, the nadroparin calcium sample solution of 5 different batches, all has obvious maximal ultraviolet absorption peak at 302 ± 1nm place.This shows, Phen-zinc sulfate system detects for the ultraviolet spectrum of different batches nadroparin calcium sample solution, has good system suitability.
Experiment two, Phen-zinc sulfate system are to the anti-interference of calcium ion
Get 7 15mL centrifuge tubes, wherein 6 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate, 1.0mL8.0mM Phen solution and 3.0mL1.0M sodium acetate solution respectively, abundant mixing. then, toward in 6 sample test systems, all add 2.00mL10% nadroparin calcium sample solution, shake up.Then, toward in 6 sample test systems, add 0.00 respectively, 0.05,0.10,0.15,0.20, the 5.0M calcium chloride solution of 0.25mL, and 3.00,2.95,2.90,2.85,2.80, the water for injection of 2.75mL; Toward in blank system, add 5.0mL water for injection, fully mix.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 6 sample test systems respectively.Gained test result as shown in Figure 4.
Can find from Fig. 4, concentration range is the additional calcium ion of 2.5%-12.5%, does not significantly affect the test result of Phen-zinc sulfate ultraviolet spectrum system.This shows, Phen-zinc sulfate ultraviolet spectrum system, has very strong Anti-Jamming to the calcium ion concentration in nadroparin calcium sample test solution.
Experiment three, sodium acetate solution consumption are on the impact of Phen-zinc sulfate ultraviolet spectrum test system
Get 7 15mL centrifuge tubes, wherein 6 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate and 1.0mL8.0mM Phen solution respectively, fully mix.Then, toward in 6 sample test systems, all add 2.00mL10% nadroparin calcium sample solution, shake up.Then, toward in 6 sample test systems, add 1.0 respectively, 1.5,2.0,3.0,3.5, the 1.0M sodium acetate solution of 4.0mL, and 5.0,4.5,4.0,3.0,2.5, the water for injection of 2.0mL; Toward in blank system, add 3.0mL sodium acetate solution and 5.0mL water for injection, fully mix.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 6 sample test systems respectively.Gained test result as shown in Figure 5.
Can find from Fig. 5, different sodium acetate solution consumptions, to the test result of Phen-zinc sulfate ultraviolet spectrum system, only can produce smaller change.This shows, the impact of sodium acetate solution consumption on Phen-zinc sulfate ultraviolet spectrum test system is very little.But, in order to strengthen Phen-zinc sulfate ultraviolet spectrum system, to the anti-interference of testing sample solution acidity and salt content, in testing program, the addition keeping sodium acetate solution is 3.0mL.
Experiment four, Phen solution usage are on the impact of Phen-zinc sulfate ultraviolet spectrum test system
Get 7 15mL centrifuge tubes, wherein 6 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate and 3.0mL sodium acetate solution respectively, fully mix.Then, toward in 6 sample test systems, all add 2.00mL10% nadroparin calcium sample solution, shake up.Then, toward in 1-6 sample test system, add 0 respectively, 0.5,0.8,1.0,1.2, the 8.0mM Phen solution of 1.5mL, and 4.0,3.5,3.2,3.0,2.8, the water for injection of 2.5mL; Toward in blank system, add 1.0mL8.0mM Phen solution and 5.0mL water for injection, fully mix.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 6 sample test systems respectively.Acquired results as shown in Figure 6.
Can find, different Phen solution usage Phen-zinc sulfate ultraviolet test system is had to the impact of highly significant from Fig. 6.When the consumption of Phen solution is less than the Phen solution usage in blank system, gained ultraviolet spectrum drops on the minus zone of absorbance; And when the consumption of Phen solution is equal to or greater than the Phen solution usage in blank system, the absorbance of gained ultraviolet spectrum at Zheng Qu, and becomes multiple to increase.Which illustrate, nadroparin calcium test sample, at the maximal ultraviolet absorption peak at 302nm place, derives from Phen and is combined while nadroparin calcium molecule, zinc ion, and be not the combination of zinc ion and nadroparin calcium molecule.
Experiment five, sodium nitrite solution consumption are on the impact of Phen-zinc sulfate ultraviolet spectrum test system
Get 7 15mL centrifuge tubes, wherein 6 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate, 1.0mL8.0mM Phen solution and 3.0mL1.0M sodium acetate solution respectively, fully mix.Then, toward in 6 sample test systems, all add 2.00mL10% nadroparin calcium sample solution, shake up.Then, toward in 1-6 sample test system, add 0.00 respectively, 0.05,0.10,0.15,0.20, the 2.0M sodium nitrite solution of 0.25mL, and 3.00,2.95,2.90,2.85,2.80, the water for injection of 2.75mL; Toward in blank system, add 5.0mL water for injection, fully mix. leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 6 sample test systems respectively.Gained test result is as shown in Figure 7.
Can find from Fig. 7, along with the increase of sodium nitrite solution consumption, gained ultraviolet spectrum fades away at the maximum absorption band at 302nm place, simultaneously at the maximum absorption band that 354nm place appearance one is new.Which illustrate, sodium nitrite solution is very large on the impact of Phen-zinc sulfate ultraviolet test system, and its maximum absorption band can be made to produce significant Red Shift Phenomena.According to the chemical structure characteristic of nadroparin calcium, its reason is likely in test system, additional a large amount of nitrite ions have oxidation, change the hydroxyl in nadroparin calcium molecule into aldehyde radical or carboxyl, thus increase the conjugative effect of colour developing group, and then its maximal ultraviolet absorption peak is made to create Red Shift Phenomena., this also illustrates, the hydroxyl in nadroparin calcium molecule take part in the formation of whole color development system meanwhile.
By above-mentioned experiment one to experiment five, can prove that Phen-zinc sulfate system can be used as measuring the concentration of nadroparin calcium solution.Test the standard working curve first setting up Phen-zinc sulfate system ultraviolet spectroscopy nadroparin calcium solution concentration, then confirm the reliability of this working curve by experiment for below two groups.
Experiment six, Phen-zinc sulfate ultraviolet spectrum test system are to the standard working curve of nadroparin calcium
Get 7 15mL centrifuge tubes, wherein 6 is sample test system, and 1 is blank system (not adding nadroparin calcium sample).First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate, 1.0mL8.0mM Phen solution and 3.0mL1.0M sodium acetate solution respectively, fully mixing (Zn in other embodiments
2+the concentration range of ion, in the whole mixed system after adding nadroparin calcium and water for injection, is 0.05 ~ 0.3mM; In other embodiments, the amount of substance of Phen is Zn
2+more than 2 times of the amount of substance of ion).Then, toward in sample test system, add the nadroparin calcium sample solution of 2.0mL variable concentrations respectively, concentration successively 5.69%, 7.86%, 9.61%, 11.84%, 14.14%, 16.33%, and 3.0mL water for injection, shake up; Toward in blank system, add 5.0mL water for injection, fully mix.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan the ultraviolet spectrum of 6 sample test systems respectively.As shown in Figure 8, in Fig. 8, the concentration of the nadroparin calcium solution added of shown curve is followed successively by gained test result from top to bottom: 5.69%, 7.86%, 9.61%, 11.84%, 14.14% and 16.33%.
Can find from Fig. 8, Phen-zinc sulfate ultraviolet test system is to nadroparin calcium sample, and within concentration range 5.69% ~ 16.33%, present good linear relationship, namely the quality of nadroparin calcium is relative to Zn
2+the amount of substance of ion, when the scope of 56.9g/mmol ~ 163.3g/mmol, can quantitatively detect exactly, and the standard working curve equation calculated is: y=2.6362x+0.0644, R
2=0.9987, wherein y represents the absorbance at 302nm place, and x represents the concentration of the nadroparin calcium added.Show according to the further result of study of the method, Phen-zinc sulfate ultraviolet test system range of linearity total to nadroparin calcium sample concentration is: 1.184% ~ 16.33%, namely when the quality of nadroparin calcium is relative to Zn
2+the amount of substance of ion, when the scope of 11.84g/mmol ~ 163.3g/mmol, is applicable to the detection of nadroparin calcium actual sample in production run.
Experiment seven, Phen-zinc sulfate ultraviolet spectrum test system are to the detection of nadroparin calcium actual sample
On the basis of experiment six, separately get 3 15mL centrifuge tubes, as actual sample test system, namely repeated test three times, then averages.First, toward in all centrifuge tubes, add 1.0mL2.0mM solution of zinc sulfate, 1.0mL8.0mM Phen solution and 3.0mL1.0M sodium acetate solution respectively, fully mix.Then, toward in actual sample test system, add nadroparin calcium solution through 2 times of dilutions that 2.0mL produces in actual production process (as shown in Figure 2, actual product is here the intermediate product solution between ultrafiltration technology and alcohol precipitation process) respectively, and 3.0mL water for injection, shake up.Leave standstill after 15 minutes, using blank system as placebo solution, within the scope of 200 ~ 400nm, scan its ultraviolet spectrum, the absorbance at record 302nm place.By the standard working curve equation that gained absorbance substitutes in experiment six, calculate its concentration, test result lists in table 1.
Table 1 Phen-zinc sulfate ultraviolet spectrum test system is to the detection of nadroparin calcium actual sample
It is pointed out that, in above-mentioned all experiments, the sodium acetate as buffer solution solute can replace with at least one in sodium acetate, sodium hydrogen phosphate, sodium dihydrogen phosphate, sodium carbonate, sodium bicarbonate and niter cake.When preparing the solution of Phen, available solute comprises at least one in water, ethanol, methyl alcohol, isopropyl alcohol, n-propanol and normal butyl alcohol.
From above-described embodiment, Phen disclosed by the invention-zinc sulfate ultraviolet spectroscopy measures the solution concentration application Phen-zinc sulfate ultraviolet spectrum test system of nadroparin calcium, achieve the Quantitative detection to nadroparin calcium solution concentration, the range of linearity of its analytical approach is 1.184% ~ 16.33%; Meanwhile, the method has good antijamming capability to calcium ion, sodion and sulfate ion, can be applied in the actual production process of nadroparin calcium medicine, carries out Concentration Testing to middle product solution or final mean annual increment solution.
It is to be noted, the method is not only applicable to the concentration determination of nadroparin calcium medicine, and the solution concentration that can be applied to other several heparin class medicine quantitatively detects, such as existing Dalteparin Sodium, Enoxaparin Sodium, Parnaparin Sodium, spit of fland liquaemin commercially, even comprises the liquaemin raw material etc. of upstream.Meanwhile, based on the test philosophy of ultraviolet spectrum, the Phen in the present invention-zinc sulfate system can be deformed into simply: Phen-copper sulphate system, Phen-ammonium ferric sulfate system, Phen-silver nitrate system etc.
Above-described embodiment is only illustrating technical scheme of the present invention, is not intended to limit the present invention, and every equivalent replacement or conversion doing same design to the present invention, all belongs to protection scope of the present invention.
Claims (8)
1. apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: under the environment of sodium acetate solution, by Zn
2+solion, Phen solution and nadroparin calcium solution mix, Zn described in mixed solution
2+the concentration range of ion is at 0.05-0.3mM, and the amount of substance of described Phen is Zn
2+more than 2 times of the amount of substance of ion, the quality of described nadroparin calcium is relative to Zn
2+the amount of substance of ion is 11.84g/mmol-163.3g/mmol, and the range of linearity is 1.184%-16.33%; The concentration of quantitative measurement nadroparin calcium is carried out by the ultraviolet absorptivity measuring mixed solution.
2. apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration according to claim 1, it is characterized in that: during Criterion working curve, add the nadroparin calcium solution for concentration known of mixed solution, and the quality of described nadroparin calcium is relative to described Zn
2+the amount of substance of ion is 11.84g/mmol-163.3g/mmol;
After Criterion working curve, add the nadroparin calcium solution for unknown concentration of mixed solution, the concentration of described nadroparin calcium solution is calculated, when the quality of the nadroparin calcium added is relative to described Zn according to the absorbance of mensuration and the standard working curve of foundation
2+the amount of substance of ion is not when the scope of 11.84g/mmol-163.3g/mmol, change by method that is concentrated or dilution the concentration adding nadroparin calcium solution in mixed solution when not changing and adding volume, measure its absorbance and utilize standard working curve to calculate the concentration of described nadroparin calcium solution, making the quality of nadroparin calcium relative to described Zn
2+the amount of substance of ion is in the scope of 11.84g/mmol-163.3g/mmol;
The concentration of described nadroparin calcium refers to the quality percent by volume of nadroparin calcium.
3. according to claim 1 or 2, apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: the solvent of described mixed solution is at least one in water, ethanol, methyl alcohol, isopropyl alcohol, n-propanol and normal butyl alcohol.
4. according to claim 1 or 2, apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: the concentration of described sodium acetate in described mixed solution is 0.3M.
5. according to claim 1 or 2, apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: described Zn
2+ion from zinc sulfate, described Zn
2+the concentration of ion is 0.2mM.
6. according to claim 1 or 2, apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: the amount of substance of described Phen is described Zn
2+4 times of the amount of substance of ion.
7. according to claim 1 or 2, apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration, it is characterized in that: when measuring the absorbance of described mixed solution, ultraviolet wavelength is 302 ± 1nm.
8. apply the method that Phen-zinc sulfate ultraviolet spectroscopy measures nadroparin calcium solution concentration according to claim 4, it is characterized in that: during preparation mixed solution, described zinc sulfate solution is 1.0mL, 2.0mM, the ethanolic solution of described Phen is 1.0mL, 8.0mM, described aqueous sodium acetate solution is 3.0mL, 1.0M, after abundant mixing, add 2.0mL nadroparin calcium solution and 3.0mL water for injection, abundant mixing also, after leaving standstill, measures the absorbance of described mixed solution at 302 ± 1nm.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102435571A (en) * | 2011-09-16 | 2012-05-02 | 东华大学 | Method for detecting heparin content with polyethyleneimine-stabilized gold nanoparticle |
| CN102721674A (en) * | 2012-05-24 | 2012-10-10 | 北京化工大学 | Quaternary fluorescence enhancement system of Eu-vanillin type Schiff base-phenanthroline-Zn, preparation method and application |
| CN102759596A (en) * | 2012-07-09 | 2012-10-31 | 山东大学 | Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050069958A1 (en) * | 2003-09-26 | 2005-03-31 | Mills Rhonda A. | Method for simultaneous evaluation of a sample containing a cellular target and a soluble analyte |
| US8716491B2 (en) * | 2006-07-18 | 2014-05-06 | New York University | Benzimidazolium dyes and their use as fluorescent chemosensors |
-
2013
- 2013-01-30 CN CN201310036199.9A patent/CN103149170B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102435571A (en) * | 2011-09-16 | 2012-05-02 | 东华大学 | Method for detecting heparin content with polyethyleneimine-stabilized gold nanoparticle |
| CN102721674A (en) * | 2012-05-24 | 2012-10-10 | 北京化工大学 | Quaternary fluorescence enhancement system of Eu-vanillin type Schiff base-phenanthroline-Zn, preparation method and application |
| CN102759596A (en) * | 2012-07-09 | 2012-10-31 | 山东大学 | Method for detecting low-molecular-weight heparin by combining ion pair reversed phase chronmatogaphy and mass spectrum |
Non-Patent Citations (4)
| Title |
|---|
| Phenanthroline-based Fluorescent Chemosensor for Selective Detection of Zn2+ in DMF Buffer Solution;Wang xiu-Li et.al.;《CHEM. RES. CHINESE UNIVERSITIES》;20101125;第26卷(第6期);第884-887页，尤其是第886页左栏3.2，附图3 * |
| Spectrofluorimetric determination of trace heparin using lomefloxacin-terbium probe;Wei Wei et.al.;《Spectrochimica Acta Part A》;20061231;第63卷;第241-246页 * |
| 茜素红S催化动力学光度法测定痕量铁(Ⅲ )及反应机理探讨;陈贤光 等;《分析化学》;20060330;第34卷(第3期);第371-374页 * |
| 银(Ⅰ)邻菲罗啉体系共振瑞利散射测定肝素钠;黄伟涛等;《西南大学学报(自然科学版)》;20110720;第33卷(第07期);第50-53页 * |
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