Stabilized nano graphene oxide and preparation method thereof under the physiological condition
Technical field
The present invention relates to a kind of nanometer carbon compound, relate in particular to and a kind ofly under physiological condition, have nano graphene oxide of good stability and preparation method thereof.
Background technology
Nano material is with a wide range of applications at biomedical sector.One of them important use be exactly its as carrier, conduct drugs to specific organ-tissue in the human body, thereby play the effect of targeted therapy.The nano material that is used for medicine-carried system must have excellent biological compatibility and lower bio-toxicity, and stable under physiological condition, does not assemble.These requirements generally realize by the nano-material surface modification.At present, surface modification comparatively commonly used is to carry out polyethyleneglycol modified to nano-material surface.The biocompatibility of the nano material of Huo Deing and the stability under the physiological condition all are significantly increased like this.But there are some shortcomings in this method, promptly polyethyleneglycol modified dose cost an arm and a leg, and the preparation cost of nano material is very high, if be applied to clinically, will certainly bring very big financial burden to the consumer.
Graphene oxide is a kind of a kind of carbonaceous new material by the tightly packed one-tenth bi-dimensional cellular of monolayer carbon atom shape lattice structure, and it can utilize commercially available block graphite to make under oxidation and ultrasonic condition.Zhi Bei graphene oxide has multiple oxy radical like this, and as carboxyl, hydroxyl, epoxy radicals etc., these hydrophilic radicals make the graphene oxide can be water-soluble, and have certain stability.By further modification, can obtain the better graphene oxide of water stability (Si Y and Samulski ET.Nano Lett., 2008,8 (6): 1679-1682).But above-mentioned graphene oxide is still relatively poor in the solution such as the stability in phosphate buffer or the cell culture medium of saliferous.Polyethyleneglycol modified report (Sun X, Liu Z, the Welsher K that obtains graphene oxide stable under the physiological condition by branch-like arranged on the document, Robinson JT, Goodwin A, Zaric S, and Dai H.Nano Res, 2008,1:203-212; Liu A, Robinson JT, Sun X, and Dai H.JAm Chem Soc, 2008,130 (33): 10876-10877).But this method cost is higher, and output is lower.Limited research and the application of Graphene in fields such as biomedicines.
Summary of the invention
For improving the stability of nano graphene oxide under physiological condition, and low-cost volume production is required to satisfy its further investigation in fields such as biological medicines, the objective of the invention is to design a kind of under physiological condition the stabilized nano graphene oxide, and provide the preparation method of this nano graphene oxide.
The technical scheme that realizes first purpose of the present invention is: stabilized nano graphene oxide under the physiological condition, it is characterized in that: this nano graphene oxide also contains acetate functional group and two kinds of hydrophilic radicals of sulfonic acid functional group by further chemical modification on original finished product basis.
Further, stabilized nano graphene oxide under the aforesaid physiological condition, its length and width are all between 20nm-100nm, and its thickness is between 1nm-2nm.In addition, by modifying carboxyl functional group and sulfonic acid functional group, make the Z electromotive force of this nano graphene oxide can reach-116mV.
The technical scheme that realizes second purpose of the present invention is:
The preparation method of stabilized nano graphene oxide under the physiological condition, adopt the Hummer method with the crystalline flake graphite oxidation, then adopt ultrasonic method that the graphene oxide sheet is peeled off, adopt the ultrasonic method of probe to be broken into nanoscale to the graphene oxide sheet again, it is characterized in that: above-mentioned nano level graphene oxide is carried out acetoxylation with the water-bath ultrasonic method modify introducing acidic group functional group, more described graphene oxide is carried out the benzene sulfonic acid modified edge and introduce sulfonic group.
Further, the preparation method of stabilized nano graphene oxide under the aforementioned physiological condition, the chemical substance that wherein is used for the acetoxylation modification is sodium chloroacetate or monoxone or other alpha-halogenated carboxylic acids compound; And the chemical substance that is used for the benzene sulfonic acid modification is sulfanilic acid, metanilic acid or 2-amino-1, the 4-benzenedisulfonic acid.
The nano graphene oxide of the present invention's design can improve the stability of graphene oxide in saline solns or cell culture medium, for nano graphene oxide provides powerful guarantee in the medical application under the physiological condition.And by the preparation method that the present invention introduces, comparing to polyethyleneglycol modified method can reduce manufacturing cost conscientiously, is beneficial to volume production.
Description of drawings
Fig. 1 is preparation method's process chart of nano graphene oxide of the present invention;
Fig. 2 is the AFM photo of nano graphene oxide of the present invention;
Fig. 3 is the infared spectrum of nano graphene oxide of the present invention.
The specific embodiment
For improving the stability of nano graphene oxide under physiological condition, and low-cost volume production is required to satisfy its further investigation in fields such as biological medicines, the invention provides a kind of nano graphene oxide that under physiological conditions such as saline solns or cell culture fluid, has enough stability, this nano graphene oxide also contains acetate functional group and the sulfonic acid official can two kinds of hydrophilic radicals of figure by further chemical modification on original finished product basis.This nano graphene oxide infared spectrum as shown in Figure 3 can prove that described acetate functional group and sulfonic acid functional group successfully are connected on the nano graphene oxide.
Below, in detail the preparation method of nano graphene oxide of the present invention is described in detail by instantiation.
Can see shown in preparation method's of the present invention as shown in Figure 1 the process chart: the preparation process of this graphene oxide roughly can be divided into three parts, comprises the preparation of nano graphene oxide, the acetoxylation modification of nano graphene oxide and the benzene sulfonic acid modification of above-mentioned nano graphene oxide.Specifically:
The preparation of the first step, nano graphene oxide:
20g potassium peroxydisulfate and 5g phosphorus pentoxide are dissolved in the 30ml concentrated sulfuric acid solution, and the temperature of solution is controlled at 60 ℃; Add 50g graphite scale 1 then, reaction 6-10h, behind the cool to room temperature, water washes neutrality, air dry powdered with it again.With above-mentioned powder, join 300ml and be pre-chilled in advance in 0 ℃ the concentrated sulfuric acid.When stirring, slowly add 40g potassium permanganate, and temperature is remained on below 20 ℃.The temperature of mixture is risen to 50 ℃, and stir 2h, add the 1000ml pure water then.Be the blocking reaction, add 2.8 liters of pure water and 100ml concentration and be 30% hydrogen peroxide.The hydrochloric acid solution of mixture filtration back with 1: 10 fully washed to remove the solution metal ion, at last that the graphite oxide product 2 of resulting pale brown look is water-soluble.With the ultrasonic 2h of above-mentioned graphite oxide, 10000 rev/mins of centrifugal 30min are to 1h, and supernatant is graphene oxide 3, further the ultrasonic graphene oxide 4 of preparing nanoscale.
The acetoxylation of second step, nano graphene oxide is modified:
Earlier 10g NaOH and 5g sodium chloroacetate are added in the 50mL water and dissolve, join then in the solution that contains 100mg nano graphene oxide 2 approximately.The ultrasonic 3h of water-bath maintains the temperature at below 50 ℃.After the reaction, the hydroxyl on the graphene oxide is replaced by the COOH of sodium chloroacetate.After the reaction, brown graphene oxide solution becomes black, illustrates that graphene oxide is by partial reduction under strong alkaline condition.
The benzene sulfonic acid modification of the 3rd step, above-mentioned nano graphene oxide:
Get the 1ml concentrated hydrochloric acid and join in the 13ml water, and control its temperature about 20 ℃.Other gets the dissolution of sodium hydroxide of 100mg aminobenzenesulfonic acid with 20ml 0.25%, and adds the 60mg natrium nitrosum under constantly stirring, and this solution is slowly splashed in the above-mentioned hydrochloric acid solution, and the control temperature is below 10 ℃.Behind the reaction 30min, to wherein adding nano graphene oxide 100mg, ice bath stirs 2h, stirred overnight at room temperature, and micromolecular compound is removed in dialysis at last, just makes stabilized nano graphene oxide 5 under the physiological condition of the present invention.
Zeta potential analysis-e/or determining result shows that the Zeta electric potential of the nano graphene oxide after above-mentioned two steps modify reaches-116mV.
Above-mentioned concrete preparation technology only provides as example, and wherein acetoxylation modification and the used chemical combination material of sulfonation modification are not limited only to material mentioned in the above-mentioned example.The chemical substance that for example is used for the carboxylic acid modification can be a sodium chloroacetate, also can be that monoxone or other have the alpha-halogenated carboxylic acids compound of identical carboxyl functional group; The chemical substance that is used for the benzene sulfonic acid modification can be sulfanilic acid, metanilic acid or 2-amino-1,4-benzenedisulfonic acid etc.
The average thickness size of the nano graphene oxide of the present invention's preparation is between 1nm-2nm, and the length and width size is between 20nm-100nm.The Z electromotive force of conventional graphene oxide is about-86mV, and the nano graphene oxide that the present invention makes contains two kinds of hydrophilic radicals---COOH functional group (acetate) and SO
3H functional group (sulfonic group), the acting in conjunction of these two kinds of groups can reach-116mV the Z electromotive force of nano graphene oxide, has good stable under physiological condition.