CN1736637A - Preparation method of monodisperse gold nanometer particle for immunological chromatography test - Google Patents
Preparation method of monodisperse gold nanometer particle for immunological chromatography test Download PDFInfo
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- CN1736637A CN1736637A CN 200510017119 CN200510017119A CN1736637A CN 1736637 A CN1736637 A CN 1736637A CN 200510017119 CN200510017119 CN 200510017119 CN 200510017119 A CN200510017119 A CN 200510017119A CN 1736637 A CN1736637 A CN 1736637A
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Abstract
The invention discloses a method for preparation of monodispersed gold nanometer particles used for detection of immunity chromatography, belonging to the technical field of preparing nanometer materials. Using sodium citrate as the reducer and reducing gold schloride in water- phase, the technique is characterized in that: the molar ratio of sodium citrate to gold schloride is among 7.0- 20: 1; the temperature of the reaction system keeps 80- 99Deg. C; the PH value of gold schloride water solution is adjusted between 2.0- 5.0, the sodium citrate is added into the solution with stirring, and the stirring continues until the color of the sol is not changeable. The method is characterized in that the operation is simple, the repeatability is easy, and the cost is low; the grain diameter of gold nanometer particle is among 20- 40nm, and the polydispersion degree decreases to among 5- 10%, which satisfy the requirements of technique of biology mark and immunity chromatography more.
Description
Technical field
The invention belongs to the nano material preparation technical field, particularly a kind of immunochromatography detects the preparation method with spherical monodisperse gold nanometer particle.
Background technology
Golden nanometer particle is as one of the most stable noble metal colloidal sol, optics and the electrical properties relevant with size with many uniquenesses.In recent years, utilizing golden nanometer particle to carry out the assembling of nanostructured, the design of molectronics device and the research of biological detection label as functional unit emerges in an endless stream.The gold-marking immunity chromatography detection technique of wherein utilizing golden nanometer particle to grow up as trace labelling thing and developer, easy and simple to handle, with low cost with it, detect fast and advantage such as sensitivity height and be subjected to people's attention.One of core technology that the gold-marking immunity chromatography detects is exactly the technology of preparing of golden nanometer particle, and preparation uniform particle diameter, pattern high-quality golden nanometer particle controlled, good stability are one of key issues in this technology industrialization.
Show that after deliberation the sensitivity that the gold-marking immunity chromatography detects is closely related with employed golden nanometer particle size, size is less than the golden nanometer particle of 10nm and the coupling efficiency height of biomolecule, but its color developing effect is relatively poor, is not easy to the perusal testing result.Large-sized golden nanometer particle color developing effect is good, but coupling efficiency is relatively poor relatively, and the golden nanometer particle of size between 20～40nm can satisfy the requirement aspect coupling efficiency and the color developing effect two preferably, and is therefore suitable.
Biomarker is aqueous phase tetra chlorauric acid (HAuCl with the preparation method that golden nanometer particle prepares generally employing at present
4) reducing process, reducing agent commonly used has natrium citricum, sodium sulfocynanate, white phosphorus, sodium borohydride, ascorbic acid, tannic acid etc., wherein is most widely used with the natrium citricum reducing agent again.This method can obtain to be of a size of the golden nanometer particle of 10～20nm effectively, seek out the particle of particle diameter between 20～40nm, can recently increase the size of particle by the mole that changes natrium citricum and tetra chlorauric acid, but increase along with size, the polydispersity of particle also increases thereupon, and particle morphology shows as ellipsoid or other is irregularly shaped, and particle stability reduces, especially in synthesizing in batches, be difficult to control the repeatability between different synthetic batch.
For overcoming these defectives, many research groups have developed some new technologies of preparing.Develop a kind of seeded growth method as Natan group and obtained monodispersed large scale golden nanometer particle (Brown, K.R.; Natan, M.J.Langmuir 1998,14, and 726.), but the particle concentration that this method obtains is very low, is unsuitable for actual production.Also have the organic ligand that utilizes mercaptan or other kinds control gold nanometer particle grain size the preparation method, but all exist reaction condition to be difficult to control, problem such as poor reproducibility and particle concentration be low as a result and be difficult to satisfy the production requirement of immuno-chromatographic assay technology.
The golden nanometer particle preparation method who is close most with the present invention is the Frens method, this method discloses the reduction reaction of utilizing natrium citricum and tetra chlorauric acid, by changing natrium citricum obtains different-grain diameter in the concentration of particle nucleation stage golden nanometer particle (13～150nm).It is that the detailed process of golden nanometer particle of 13nm is as follows that the Frens legal system is equipped with particle diameter: at first, get 5ml concentration and be 1% HAuCl
4Solution is diluted with water to 100ml, is heated to boiling, at the sodium citrate solution 5ml of stirring condition property adding next time 1%, this moment natrium citricum and Au
3+Mol ratio be about 3.5: 1, keep boiling 5 minutes, can obtain red golden nanometer particle colloidal sol.This moment, the average grain diameter of golden nanometer particle was 13nm, and the particle polydispersity is 9.8%, and it is spherical that particle is essentially.In the Frens method, the golden nanometer particle of different-grain diameter can obtain by identical preparation process, only needs to change the amount that sodium citrate solution adds, and promptly changes natrium citricum and Au
3+Mol ratio, both can obtain the golden nanometer particle of different-grain diameter.In general, the particle diameter of the golden nanometer particle of preparation is along with natrium citricum and Au
3+Reducing of mol ratio and increase gradually.As preparing the golden nanometer particle that particle diameter is 20～40nm, need be with natrium citricum and Au
3+Mol ratio be reduced in 2.7～1.4: between 1, natrium citricum and Au
3+The decline that reduces to have caused the particle monodispersity of mol ratio.As particle diameter is that its polydispersity of golden nanometer particle of 20nm is 12.9%, and particle diameter is that its polydispersity of golden nanometer particle of 28nm is 20.9%, and particle diameter is that its polydispersity of golden nanometer particle of 35nm is 32%.And the golden nanometer particle of Frens method preparation is along with the increase of size, and its shape becomes ellipse or other are irregularly shaped from sphere, can't satisfy the application request of immuno-chromatographic assay technology.
Summary of the invention
The technical problem to be solved in the present invention is, at problem and the defective in the present golden nanometer particle aqueous phase preparation method, set up a kind of particle diameter method between 20～40nm, monodispersed color of spherical gold for preparing, that this method has is easy and simple to handle, good reproducibility, with low cost, the characteristics that are suitable for producing, prepared gold nanometer particle grain size controllability and monodispersity are good, are applicable to the needs of immuno-chromatographic assay technology.
The preparation method of the monodispersed golden nanometer particle of sphere of the present invention is, utilizes natrium citricum to reduce tetra chlorauric acid (HAuCl at aqueous phase
4), by improving natrium citricum and Au
3+Molar ratio keep the monodispersity of nano particle, thereby control the spherical monodisperse gold nanometer particle of the speed preparation different-grain diameter of nucleation and growth in the reaction by the pH value of regulation and control reaction system.
Concrete is, makees reducing agent with natrium citricum, at aqueous phase reduction tetra chlorauric acid, it is characterized in that the mol ratio of natrium citricum and tetra chlorauric acid is 7.0～20: 1 scope; Reaction system keeps 80～99 ℃; Course of reaction is that the tetra chlorauric acid pH value of aqueous solution is adjusted between 2.0～5.0, injects natrium citricum under stirring condition, continues stirring reaction to the colloidal sol constant color.
The concentration range of the tetra chlorauric acid aqueous solution can be 0.05 * 10
-3～1.0 * 10
-3Mol/L.
The regulation and control of said tetra chlorauric acid pH value of aqueous solution are, under the mol ratio of natrium citricum of determining and tetra chlorauric acid, by adding acid, alkali and being equivalent to acid or the salt of alkali is adjusted to 2.0～5.0 with the tetra chlorauric acid pH value of aqueous solution.Reasonable pH value scope is 2.9～3.6.
That method of the present invention has is easy and simple to handle, good reproducibility, with low cost, the characteristics that are suitable for producing, prepared gold nanometer particle grain size polydispersity is reduced between 5～10% by 10～35% in the Frens method, particle diameter can be regulated and control between 20～40nm continuously, and the sample resting period is longer.The golden nanometer particle that these advantages are synthesized is more suitable for the requirement of biomarker and immunochromatography technique.
The specific embodiment:
Embodiment 1:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing hydrochloric acid solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 2.9, this solution is heated to 99 ℃, at the sodium citrate solution 5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1, continues to stir and keep 99 ℃ of solution, do not change to colloidal sol color (redness), promptly obtaining average grain diameter is 20.0nm, and the particle polydispersity is 6.0%, is spherical golden nanometer particle substantially.
Embodiment 2:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing sodium hydroxide solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 3.3, this solution is heated to 99 ℃ on the magnetic force heating stirrer, at the sodium citrate solution 5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1, continues to stir and maintain the temperature at 99 ℃, do not change until colloidal sol color (redness), promptly obtaining average grain diameter is 26.4nm, and the particle polydispersity is 3.7%, is spherical golden nanometer particle substantially.
Embodiment 3:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing hydrochloric acid solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 3.0, this solution is heated to 99 ℃ on the magnetic force heating stirrer, at the sodium citrate solution 7.5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 10.5: 1; Continue to stir and also to keep 99 ℃ of reaction temperatures, do not change to colloidal sol color (redness), promptly obtaining average grain diameter is 31.8nm, and the particle polydispersity is 6.4%, be the golden nanometer particle of sphere substantially.
Embodiment 4:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing solution of potassium carbonate (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 3.6, this solution is heated to 99 ℃ on the magnetic force heating stirrer, at the sodium citrate solution 5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1; Continue to stir and also to keep 99 ℃ of reaction temperatures, do not change to colloidal sol color (redness), promptly obtaining average grain diameter is 36.1nm, and the particle polydispersity is 6.7%, be the golden nanometer particle of sphere substantially.
Embodiment 5:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilize sodium hydroxide solution (1mol/L) to regulate tetra chlorauric acid pH value of aqueous solution 3.3, this solution is heated to 99 ℃ on the magnetic force heating stirrer, sodium citrate solution 12.5ml in stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 17.5: 1, continue to stir and keep 99 ℃ of reaction temperatures, do not change until colloidal sol color (redness), promptly obtaining average grain diameter is 38.5nm, the particle polydispersity is 7.8%, is spherical golden nanometer particle substantially.
Embodiment 6:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing sodium hydroxide solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 3.3, this solution is heated to 90 ℃ on the magnetic force heating stirrer, at the sodium citrate solution 5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1, continues to stir and keep 90 ℃ of reaction temperatures, do not change until colloidal sol color (redness), promptly obtaining average grain diameter is 24.2nm, and the particle polydispersity is 6.1%, is spherical golden nanometer particle substantially.
Embodiment 7:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing sodium hydroxide solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 3.3, this solution is heated to 80 ℃ on the magnetic force heating stirrer, at the sodium citrate solution 5ml of stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1, continues to stir and keep 80 ℃ of reaction temperatures, do not change until colloidal sol color (redness), promptly obtaining average grain diameter is 24.9nm, and the particle polydispersity is 7.1%, is spherical golden nanometer particle substantially.
Embodiment 8:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilizing hydrochloric acid solution (1mol/L) to regulate the tetra chlorauric acid pH value of aqueous solution is 2.0, this solution is heated to 90 ℃, sodium citrate solution 10ml in stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 14: 1, continue to stir and keep 90 ℃ of temperature, do not change until colloidal sol color (redness).The golden nanometer particle average grain diameter of preparation is 24.0nm, and the particle polydispersity is 7.3%, and particle is spherical substantially.
Embodiment 9:
Getting 500ml concentration is 0.25 * 10
-3The HAuCl of mol/L
4Solution, utilize sodium hydroxide solution (1mol/L) to regulate tetra chlorauric acid pH value of aqueous solution 5.0, this solution is heated to 99 ℃ on the magnetic force heating stirrer, sodium citrate solution 5ml in stirring condition property adding next time 5%, the mol ratio of natrium citricum and tetra chlorauric acid is about 7: 1, continue to stir and keep boiling, do not change until colloidal sol color (redness).The golden nanometer particle average grain diameter of preparation is 35.5nm, and the particle polydispersity is 7.2%, and particle is spherical substantially.
In the above embodiments 1～9, HAuCl
4The concentration of solution can be 0.05 * 10
-3～1.0 * 10
-3Change in the mol/L scope, the effect that makes golden nanometer particle do not had too much influence, promptly the particle diameter of golden nanometer particle still 20～40nm scope, polydispersity still between 5～10%, shape of particle be sphere substantially.
Claims (2)
1, a kind of preparation method of monodisperse gold nanometer particle for immunological chromatography test, make reducing agent with natrium citricum,, it is characterized in that at aqueous phase reduction tetra chlorauric acid, the mol ratio of natrium citricum and tetra chlorauric acid is 7.0～20: 1 scope, and reaction system keeps 80～99 ℃; Course of reaction is that the tetra chlorauric acid pH value of aqueous solution is adjusted between 2.0～5.0, injects natrium citricum under stirring condition, continues stirring reaction to the colloidal sol constant color.
According to the preparation method of the described a kind of monodisperse gold nanometer particle for immunological chromatography test of claim 1, it is characterized in that 2, the concentration range of the tetra chlorauric acid aqueous solution is 0.05 * 10
-3～1.0 * 10
-3Mol/L; The tetra chlorauric acid pH value of aqueous solution is adjusted between 2.9～3.6.
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| CN100463747C (en) * | 2006-09-30 | 2009-02-25 | 南京大学 | Method for preparing multiple high-purity anisotropic gold nano particle |
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