CN106581697A - Nanometer contrast agent with lung-cancer-targeting magnetic resonance imaging capacity and preparation method of nanometer contrast agent - Google Patents
Nanometer contrast agent with lung-cancer-targeting magnetic resonance imaging capacity and preparation method of nanometer contrast agent Download PDFInfo
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Abstract
The invention relates to a nanometer contrast agent with the lung-cancer-targeting magnetic resonance imaging capacity and a preparation method of the nanometer contrast agent, belonging to the technical field of radiodiagnostics. The preparation method of the nanometer contrast agent comprises the following steps: preparing asymmetrical Au-Ni nano-particles, preparing asymmetrical Au-Ni nano-particles with two modified ends, and preparing the nanometer contrast agent. The prepared nanometer contrast agent with the lung-cancer-targeting magnetic resonance imaging capacity has the good morphological control capacity, good biocompatibility and excellent lung cancer targeting property, can be well dispersed in a biological vector, and has the excellent magnetic resonance targeting imaging capacity for lung cancer cells. In addition, the preparation method provided by the invention is easy and convenient to operate, the adopted raw materials are easily available, and the cost is low.
Description
Technical field
The invention belongs to radiodiagnostics technical field, and in particular to a kind of with pulmonary carcinoma targeting nuclear magnetic resonance ability
Nano-contrast agent and preparation method thereof.
Background technology
Pulmonary carcinoma is one of common malignant tumor of China, and pulmonary carcinoma early stage lacks distinctive clinical manifestation, and some patientss exist
In the middle and advanced stage of disease when disease is made a definite diagnosis.Hence set up a kind of more effective system to realize that the early diagnosiss to pulmonary carcinoma show
Obtain particularly important.The contrast agent of nuclear magnetic resonance (magnetic resonance imaging, MRI) at present is superparamagnetism
Contrast agent and Water-soluble paramagnetic contrast agent.Such as superparamagnetism Fe3O4Preparation pairT 2There is significant impact time, and it is led
Have the disadvantage under dimmed imaging pattern, to obtain more dark-coloured image, the contrast of organ, imaging of tissue to target site
Degree is little, and easily mutually obscures with the MRI effects of surrounding tissue and cause mistaken diagnosis.It is different, general paramagnetic contrast medium
It is main to affectT 1Time, that is, longitudinal relaxation time is shortened by, is madeT 1Weighting is as brightening, therefore paramagnetic contrast medium（T 1Radiography
Agent）Typically all just strengthen contrast agent, easily clearly distinguishing lesions position with normal structure, and then strengthen medical diagnosis on disease
Effect.Licensed listing at present is most by ferriferrous oxide particles core with the superparamagnetic contrast agents just in clinical experiment
The one floor height polymers composition of the heart and outside cladding, particle diameter is about 25-40 nanometers, and its synthesis technique is complicated, manufacturing cost is high, grain
Footpath is larger, degree of crystallinity is low, MRI imaging effects are poor, surface coating layer uneven thickness.And presently used paramagnetic contrast medium
The overwhelming majority is the organic coordination compound containing small molecule gadolinium, is extracellular reagent, in vivo in non-specific distribution, it is adaptable to
The imaging of brain, kidney and blood system, but it is poor to the imaging effect of the systems such as transhepatic cholangiography, and this causes them to cannot act as selectivity
(targeting) contrast agent.
Folacin receptor is a kind of cell surface protein that can be used for tumor marker, with useful antitumor drug transmission target
Position.Therefore Folic Acid is often connected low-molecular-weight drug molecule or the high polymer end of the chain, transmits molecule that targeting combines to pernicious
Tumor surface, plays the purpose that targeting recognizes tumor.Folacin receptor can show in lung carcinoma cell in overexpression, therefore pulmonary carcinoma
Write the intake ability for lifting the nano-particle crossed to modified with folic acid.
The content of the invention
The purpose of the present invention is to overcome the deficiencies in the prior art and provide a kind of with pulmonary carcinoma targeting nuclear magnetic resonance ability
Nano-contrast agent and preparation method thereof, should have nano-contrast agent of pulmonary carcinoma targeting nuclear magnetic resonance ability will be with pulmonary carcinoma target
Asymmetric golden nickel is anchored to the small molecule Folic Acid of identification ability（Au-Ni）Nano grain surface, to realize it to lung carcinoma cell
Targeting imaging.
The present invention is employed the following technical solutions：
The preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability, comprises the steps：
Step one：A certain amount of nickel acetylacetonate is placed in the mixed solvent of oleyl amine and 18-amine., is heated to treating that it is completely molten
Xie Shi, the HAuCl of Deca same molar4, intensification stirring reaction；Reaction treats that system is lowered the temperature after terminating, add chloroform, then side to stir
Mix side and add appropriate 18-amine., permanent magnet is placed outside reaction vessel, collect asymmetric Au-Ni nano-particle, after it is anti-with chloroform
Multiple drip washing removes unnecessary nanometer gold, finally collects and obtains pure higher asymmetric Au-Ni nano-particle；
Step 2：The asymmetric Au-Ni nano-particle freeze-dried back that step one is obtained, weighs after lyophilization not
Symmetrical Au-Ni nano-particle ultrasonic disperse adds in dimethylformamide (DMF) and the mixed solution of chloroform after being completely dispersed
Enter COOH-PEG-NH2, concussion；
Step 3：SH-PEG is added, is shaken after ultrasound；
Step 4：Using Magnet collecting reaction product, and chloroform, ethanol and distilled water wash are used for several times respectively, by obtain two
The asymmetric Au-Ni nano-particle respectively through modification is held to be scattered in DMSO solution；
Step 5：During Folic Acid (FA) is dispersed in into DMSO, 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides are subsequently adding
Hydrochlorate (EDC) and N- hydroxysuccinimides (NHS), stirring reaction；
Step 6：Step 4 and step 5 gained reactant liquor are merged, after ultrasonic reaction terminates, precipitation is collected by centrifugation, be used in combination
DMSO, ethanol and distilled water protection washing for several times, that is, obtains the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability
PEG-SH@Au-Ni@COOH-PEG-FA, are scattered in stand-by in PBS.
Further, the mass ratio of oleyl amine described in step one and 18-amine. is 1:2, the mixing of oleyl amine and 18-amine. is molten
Liquid and the nickel acetylacetonate and HAuCl that add4The mass ratio of total amount is 100:0.6.
Further, heating rate when heating up described in step one is 5 DEG C/min, and intensification stirring reaction is specially：
200-210 DEG C is warming up to, stirring reaction 1-1.5 h.
Further, the mixed solution of dimethylformamide described in step 2 and chloroform and asymmetric Au-Ni nanometers
The mass ratio of granule is 100:0.02, the asymmetric Au-Ni nano-particle and COOH-PEG-NH2Mass ratio be 5:1, institute
The mass ratio for stating dimethylformamide and chloroform is 1:1, the concussion is specially shakes 3-5 h on oscillator.
Further, shake after ultrasound described in step 3 and be specially：Ultrasonic 0.5-1 h shake 3- on agitator
5 h, SH-PEG and the mass ratio of the asymmetric Au-Ni nano-particle of addition in step 2 of addition are 20:3.
Further, Folic Acid described in step 5,1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides,
Two ends described in N- hydroxysuccinimides, step 4 are respectively through the mass ratio of the asymmetric Au-Ni nano-particle of modification
5:3:3:27, stirring reaction is specially described in step 5：The stirring reaction 6-8 h at 25-28 DEG C.
Further, ultrasonic reaction time described in step 6 is 24-26 h.
The present invention also provides a kind of preparation by the described nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability
The obtained nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability of method.
The present invention compared with prior art, has the advantages that：It is of the invention obtained with pulmonary carcinoma targeting magnetic resonance
The nano-contrast agent of imaging capability has good morphology control ability, good biocompatibility and excellent pulmonary carcinoma targeting
Property, can well disperse in vector, and there is good magnetic resonance targeted imaging ability to lung carcinoma cell.In addition,
Preparation method of the invention is easy to operate, raw material that is adopting is easy to get and with low cost.
Description of the drawings
Fig. 1 is electron microscope of the asymmetric Au-Ni nano-particle of the invention in chloroform；
Fig. 2 is electron microscope of nano-contrast agent of the present invention with pulmonary carcinoma targeting nuclear magnetic resonance ability in pure water；
Fig. 3 is nano-contrast agent of the present invention with pulmonary carcinoma targeting nuclear magnetic resonance ability to human lung carcinoma cell (A549) and people
The cell survival rate of bronchiolar epithelium like cell (BEAS-2B)；
Fig. 4 is magnetic resonance targeting of nano-contrast agent of the present invention with pulmonary carcinoma targeting nuclear magnetic resonance ability to lung carcinoma cell
Image.
Specific embodiment
The present invention is described in further detail below by specific embodiment, but those skilled in the art will manage
Solution, the following example is merely to illustrate the present invention, and should not be taken as limiting the scope of the invention.The raw material examination that the present invention is used
Agent, if no special instructions, can be by commercially available acquisition.
Embodiment 1
The preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability, comprises the following steps that：
Step one：A certain amount of nickel acetylacetonate is placed in into the mixed solvent of oleyl amine and 18-amine.（The quality of oleyl amine and 18-amine.
Than for 1:2）In, be heated to 120 DEG C whne its be completely dissolved into green viscous liquid when, Deca same molar
HAuCl4, high reaction temperature is risen to 210 DEG C and magnetic agitation 1h with the heating rate of 5 DEG C/min, wherein oleyl amine and 18-amine.
Mixed solvent and the total metal content for adding（Nickel acetylacetonate and HAuCl4）Mass ratio be 100:0.6；Reaction treats body after terminating
It is that temperature is dropped to after room temperature, adds substantial amounts of chloroformic solution, reactant and product is dissolved in chloroform, mends under stirring
Plus appropriate 18-amine. does not adsorb unnecessary nano-particle up on magneton, and a block permanent magnet is placed outside reaction vessel,
Asymmetric Au-Ni nano-particle is collected by the action of a magnetic field, by can will be unnecessary single with the method for chloroform drip washing repeatedly
The nanometer gold of component is removed totally, is finally collected and is obtained the higher asymmetric Au-Ni nano-particle of purity, its pattern such as Fig. 1 institutes
Show.
Asymmetric Au-Ni nano-particle obtained in the method compare document report method (DS. Wang, Y.D. Li,
“One-Pot Protocol for Au-Based Hybrid Magnetic Nanostructures via a Noble-
Metal-Induced Reduction Process ", J.AM.CHEM.SOC.2010,132,6280-6281) obtained in nanometer
Granule, its product morphology is more controllable and product is purer, it is to avoid unnecessary Au nano-particle is to follow-up function band
The impact for coming.Now the asymmetric Au-Ni nano-particle due to by 18-amine. protection can in chloroform solvent fine dispersion, but
Dispersibility in water is poor.
Step 2：After by obtained asymmetric Au-Ni nano-particle lyophilization in step one, 100mg is weighed asymmetric
Au-Ni nano-particle ultrasonic disperse is in 500g dimethylformamides (DMF) and the mixed solution of chloroform（Dimethylformamide and chlorine
Imitative mass ratio is 1:1）In, the COOH-PEG-NH of 20mg is added in system after being completely dispersed2, by reaction unit as
3h is shaken on oscillator；
Step 3：15mg SH-PEG are added in step 2 system, is placed in after ultrasonic 0.5h on agitator and is shaken 3h；
Step 4：Using Magnet collecting reaction product, and respectively with chloroform, ethanol and distilled water wash for several times afterwards by 135mg two
The asymmetric Au-Ni nano-particle respectively through modification is held to be scattered in 20 mL DMSO solutions；
Step 5：10mg Folic Acid (FA) is dispersed in 20 mL DMSO, 1- (the 3- dimethylaminos third of 6.1 mg are subsequently adding
Base) -3- ethyl-carbodiimide hydrochlorides (EDC) and 6.3 mg N- hydroxysuccinimides (NHS), stir anti-at 25 DEG C
Answer 6h；
Step 6：Merge by step 4 and with the reactant liquor in step 5, after ultrasonic reaction 24h, precipitation is collected by centrifugation, be used in combination
DMSO, ethanol and distilled water protection washing for several times, obtains the nano-contrast agent PEG- with pulmonary carcinoma targeting nuclear magnetic resonance ability
SH@Au-Ni@COOH-PEG-FA, are scattered in stand-by in PBS, and its pattern in water is as shown in Figure 2.
Embodiment 2
The preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability, comprises the following steps that：
Step one：A certain amount of nickel acetylacetonate is placed in into the mixed solvent of oleyl amine and 18-amine.（The quality of oleyl amine and 18-amine.
Than for 1:2）In, be heated to 120 DEG C whne its be completely dissolved into green viscous liquid when, Deca same molar
HAuCl4, high reaction temperature is risen to 200 DEG C and magnetic agitation 1.5h, wherein oleyl amine and 18-amine. with the heating rate of 5 DEG C/min
Mixed solvent with add total metal content（Nickel acetylacetonate and HAuCl4）Mass ratio be 100:0.6；Reaction is treated after terminating
System temperature is dropped to after room temperature, adds substantial amounts of chloroformic solution, reactant and product is dissolved in chloroform, under stirring
Appropriate 18-amine. is added until not adsorbing unnecessary nano-particle on magneton, one block of permanent magnetism is placed outside reaction vessel
Ferrum, by the action of a magnetic field asymmetric Au-Ni nano-particle is collected, by can be by unnecessary list with the method for chloroform drip washing repeatedly
The nanometer gold of one component is removed totally, is finally collected and is obtained the higher asymmetric Au-Ni nano-particle of purity.
Step 2：After by obtained asymmetric Au-Ni nano-particle lyophilization in step one, 100mg is weighed asymmetric
Au-Ni nano-particle ultrasonic disperse is in 500g dimethylformamides (DMF) and the mixed solution of chloroform（Dimethylformamide and chlorine
Imitative mass ratio is 1:1）In, the COOH-PEG-NH of 20mg is added in system after being completely dispersed2, by reaction unit as
5h is shaken on oscillator；
Step 3：15mg SH-PEG are added in step 2 system, is placed in after ultrasonic 1h on agitator and is shaken 5h；
Step 4：Using Magnet collecting reaction product, and respectively with chloroform, ethanol and distilled water wash for several times afterwards by 135mg two
The asymmetric Au-Ni nano-particle respectively through modification is held to be scattered in 20 mL DMSO solutions；
Step 5：10mg Folic Acid (FA) is dispersed in 20 mL DMSO, 1- (the 3- dimethylaminos third of 6.1 mg are subsequently adding
Base) -3- ethyl-carbodiimide hydrochlorides (EDC) and 6.3 mg N- hydroxysuccinimides (NHS), stir anti-at 26 DEG C
Answer 7h；
Step 6：Merge by step 4 and with the reactant liquor in step 5, after ultrasonic reaction 25h, precipitation is collected by centrifugation, be used in combination
DMSO, ethanol and distilled water protection washing for several times, obtains the nano-contrast agent PEG- with pulmonary carcinoma targeting nuclear magnetic resonance ability
SH@Au-Ni@COOH-PEG-FA, are scattered in stand-by in PBS.
Embodiment 3
The preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability, comprises the following steps that：
Step one：A certain amount of nickel acetylacetonate is placed in into the mixed solvent of oleyl amine and 18-amine.（The quality of oleyl amine and 18-amine.
Than for 1:2）In, be heated to 120 DEG C whne its be completely dissolved into green viscous liquid when, Deca same molar
HAuCl4, high reaction temperature is risen to 205 DEG C and magnetic agitation 1h with the heating rate of 5 DEG C/min, wherein oleyl amine and 18-amine.
Mixed solvent and the total metal content for adding（Nickel acetylacetonate and HAuCl4）Mass ratio be 100:0.6；Reaction treats body after terminating
It is that temperature is dropped to after room temperature, adds substantial amounts of chloroformic solution, reactant and product is dissolved in chloroform, mends under stirring
Plus appropriate 18-amine. does not adsorb unnecessary nano-particle up on magneton, and a block permanent magnet is placed outside reaction vessel,
Asymmetric Au-Ni nano-particle is collected by the action of a magnetic field, by can will be unnecessary single with the method for chloroform drip washing repeatedly
The nanometer gold of component is removed totally, is finally collected and is obtained the higher asymmetric Au-Ni nano-particle of purity.
Step 2：After by obtained asymmetric Au-Ni nano-particle lyophilization in step one, 100mg is weighed asymmetric
Au-Ni nano-particle ultrasonic disperse is in 500g dimethylformamides (DMF) and the mixed solution of chloroform（Dimethylformamide and chlorine
Imitative mass ratio is 1:1）In, the COOH-PEG-NH of 20mg is added in system after being completely dispersed2, by reaction unit as
4h is shaken on oscillator；
Step 3：15mg SH-PEG are added in step 2 system, is placed in after ultrasonic 1h on agitator and is shaken 3h；
Step 4：Using Magnet collecting reaction product, and respectively with chloroform, ethanol and distilled water wash for several times afterwards by 135mg two
The asymmetric Au-Ni nano-particle respectively through modification is held to be scattered in 20 mL DMSO solutions；
Step 5：10mg Folic Acid (FA) is dispersed in 20 mL DMSO, 1- (the 3- dimethylaminos third of 6.1 mg are subsequently adding
Base) -3- ethyl-carbodiimide hydrochlorides (EDC) and 6.3 mg N- hydroxysuccinimides (NHS), stir anti-at 28 DEG C
Answer 6h；
Step 6：Merge by step 4 and with the reactant liquor in step 5, after ultrasonic reaction 26h, precipitation is collected by centrifugation, be used in combination
DMSO, ethanol and distilled water protection washing for several times, obtains the nano-contrast agent PEG- with pulmonary carcinoma targeting nuclear magnetic resonance ability
SH@Au-Ni@COOH-PEG-FA, are scattered in stand-by in PBS.
Obtained to embodiment 1 have pulmonary carcinomaT 1The nano-contrast agent of targeting nuclear magnetic resonance ability carries out biological safety
Test：By the good human lung carcinoma cell of growth conditions (A549) and human bronchial epithelial like cell (BEAS-2B) adjustment density
For 0.8-1.0 × 105Individual/mL, per the μ L of hole 100 96 well culture plates are inoculated in, 37 DEG C, 5%CO2Culture 24 is little under the conditions of saturated humidity
When after cell attachment, in cell add PEG-SH@Au-Ni@COOH-PEG-FA nano-particle so as to concentration is followed successively by：
12.5th, 25,50,100,200 μ g/ mL, each concentration acts on respectively 24 h.Per group has blank, and each Kong Jun does 3
Repeating hole；After effect terminates, supernatant culture fluid is abandoned, washed 2 times with the phosphate buffer of 200 μ L pH 7.2 per hole, per hole
Add the sodium acetate buffer (pH 5. 0) of the M of 100 μ L 0.1,0.1 % Triton X-100（Polyethylene Glycol octyl group benzene
Base ether）And 5 mM p-nitrophenyl phosphates (p-NPP) put and cultivate in 37 C incubators 2 h, experiment adds 10 when terminating
L 1 M NaOH；After culture terminates, VIVTOR is used3The detection of 1420-050 types microplate reader is per suction of the hole at 405 nm wavelength
Shading value (A), the solution of p-NPP substrate hydrolysis calculates survival rate as blank according to following formula using in not celliferous holeIC 50Value, survival rate（%）= (Adrug-blank /Acontrol-blank× 100)×100 %.Exercising result（As shown in Figure 3）Show
The obtained nano-contrast agent PEG-SH@Au-Ni@COOH-PEG-FA with pulmonary carcinoma targeting nuclear magnetic resonance ability of the invention have
There is good biological safety, to normal cell and lung carcinoma cell without inhibition in the range of regulation using dosage.
Tested with the external magnetic resonance radiography of human lung carcinoma cell (A549) and human bronchial epithelial like cell (BEAS-2B)
MR imaging effects on inspection PEG-SH@Au-Ni@COOH-PEG-FA nano-contrast agents.The sample P EG-SH@Au- of Example 1
The mg of Ni@COOH-PEG-FA 10.26 are dissolved in the ultra-pure water of 5 mL the solution for preparing that Ni concentration is 1.865 mM, then
Each 1.5 mL of solution that Ni concentration is 0.028,0.052,0.095,0.128,0.563,0.819 mM is diluted to respectively；With
0.5T MR testers test each sampleT 1Value.Figure 4, it is seen that increasing with Ni ion concentrations, A549
With BEAS-2B cell samplesT 1Signal all presents the trend for gradually becoming strong.Further, since the Folic Acid on lung carcinoma cell surface
Receptor is in that the PEG-SH@Au-Ni@COOH-PEG-FA ratios in high expression status, certain hour into lung carcinoma cell enter normal thin
Born of the same parents' is more, thus with the increase of concentration, lung carcinoma cellT 1Wanting for signal enhancing Amplitude Ratio normal cell is big, realizes to lung
The targeting imaging of cancerous cell.
Claims (8)
1. there is the preparation method of the nano-contrast agent of pulmonary carcinoma targeting nuclear magnetic resonance ability, it is characterised in that including following step
Suddenly：
Step one：A certain amount of nickel acetylacetonate is placed in the mixed solvent of oleyl amine and 18-amine., is heated to treating that it is completely molten
Xie Shi, the HAuCl of Deca same molar4, intensification stirring reaction；Reaction treats that system is lowered the temperature after terminating, add chloroform, then side to stir
Mix side and add appropriate 18-amine., permanent magnet is placed outside reaction vessel, collect asymmetric Au-Ni nano-particle, after it is anti-with chloroform
Multiple drip washing removes unnecessary nanometer gold, finally collects and obtains pure higher asymmetric Au-Ni nano-particle；
Step 2：The asymmetric Au-Ni nano-particle freeze-dried back that step one is obtained, weighs after lyophilization not
Symmetrical Au-Ni nano-particle ultrasonic disperse is added in the mixed solution of dimethylformamide and chloroform after being completely dispersed
COOH-PEG-NH2, concussion；
Step 3：SH-PEG is added, is shaken after ultrasound；
Step 4：Using Magnet collecting reaction product, and chloroform, ethanol and distilled water wash are used for several times respectively, by obtain two
The asymmetric Au-Ni nano-particle respectively through modification is held to be scattered in DMSO solution；
Step 5：Folic Acid is dispersed in DMSO, 1- (3- dimethylamino-propyls) -3- ethyl carbodiimide hydrochlorides are subsequently adding
Salt and N- hydroxysuccinimides, stirring reaction；
Step 6：Step 4 and step 5 gained reactant liquor are merged, after ultrasonic reaction terminates, precipitation is collected by centrifugation, be used in combination
DMSO, ethanol and distilled water protection washing for several times, that is, obtains the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability
PEG-SH@Au-Ni@COOH-PEG-FA, are scattered in stand-by in PBS.
2. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, the mass ratio of oleyl amine described in step one and 18-amine. is 1:2, the mixed solution and addition of oleyl amine and 18-amine.
Nickel acetylacetonate and HAuCl4The mass ratio of total amount is 100:0.6.
3. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, heating rate when heating up described in step one is 5 DEG C/min, intensification stirring reaction is specially：It is warming up to 200-
210 DEG C, stirring reaction 1-1.5 h.
4. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, the quality of the mixed solution of dimethylformamide described in step 2 and chloroform and asymmetric Au-Ni nano-particle
Than for 100:0.02, the asymmetric Au-Ni nano-particle and COOH-PEG-NH2Mass ratio be 5:1, the dimethyl methyl
The mass ratio of amide and chloroform is 1:1, the concussion is specially shakes 3-5 h on oscillator.
5. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, shakes after ultrasound described in step 3 and be specially：Ultrasonic 0.5-1 h shake 3-5 h on agitator, addition
SH-PEG is 3 with the mass ratio of the asymmetric Au-Ni nano-particle of addition in step 2:20.
6. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, Folic Acid described in step 5,1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, N- hydroxyls fourth two
Two ends described in acid imide, step 4 are 5 respectively through the mass ratio of the asymmetric Au-Ni nano-particle of modification:3:3:27, step
Stirring reaction is specially described in rapid five：The stirring reaction 6-8 h at 25-28 DEG C.
7. the preparation method of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability according to claim 1, its
It is characterised by, ultrasonic reaction time described in step 6 is 24-26 h.
8. the preparation of the nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability by described in any one of claim 1 to 7
The obtained nano-contrast agent with pulmonary carcinoma targeting nuclear magnetic resonance ability of method.
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