CN112516972A - Nano material capable of selectively enriching and separating sulfur-containing compounds, and preparation method and application thereof - Google Patents
Nano material capable of selectively enriching and separating sulfur-containing compounds, and preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a nano material capable of selectively enriching and separating sulfur-containing compounds, which is a core-shell ferroferric oxide/polydopamine/gold (Fe3O4/PDA/Au) composite magnetic nano material. The preparation method of the material comprises the following steps: firstly, preparing Fe3O4/PDA modified by amino, reducing chloroauric acid (HAuCl4) by tetrakis (hydroxymethyl) phosphonium chloride (THPC) to obtain gold nanoparticles with small particle size, and adsorbing the gold nanoparticles on the surface of the Fe3O4/PDA through electrostatic interaction and the viscosity of the PDA. And finally, gold nanoparticles adsorbed on the surface of Fe3O4/PDA are used as crystal nuclei, and HAuCl4 is reduced for multiple times by hydroxylamine hydrochloride to form a complete gold nano layer. The core-shell type Fe3O4/PDA/Au composite nano material prepared by the method has good dispersibility, stability and biocompatibility, and can effectively and selectively enrich and separate sulfur-containing compounds from a sample solution.
Description
Technical Field
The invention relates to a core-shell type magnetic nano material capable of selectively enriching and separating sulfur-containing compounds, a preparation method and application thereof, and Fe of the invention3O4the/PDA/Au composite material can be used for enriching sulfur-containing compounds in the fields of biology, chemistry, medicine and the likeAnd (5) separating.
Background
Sulfur-containing compounds widely exist in nature and in organisms, such as cysteine, glutathione and homocysteine, participate in metabolism in vivo, and have important physiological functions. The level of sulfhydryl compounds in organisms is closely related to clinical diseases, such as liver injury, skin diseases and the like caused by the lack of cysteine in the organisms, the higher homocysteine can cause neurological diseases, Alzheimer's disease, cardiovascular diseases and the like, and glutathione is taken as the sulfhydryl compound with the highest intracellular content, is an important antioxidant in the organisms, can clear excessive free radicals in the organisms and plays an important role in maintaining normal life activities of the organisms. Therefore, the method has important significance for monitoring the sulfur-containing compounds. However, the substrate environment of the sulfur-containing compounds is complex, the interference is more, and the content of many sulfur-containing compounds in the substrate is very low, so that the sulfur-containing compounds can be effectively enriched, the interference can be reduced, and the detection sensitivity and accuracy can be improved.
The neuropeptide components in animal bodies and polypeptide toxin components in some animals, wherein the polypeptide containing disulfide bonds usually shows strong activity or strong toxicity due to the stable spatial structure, and is one of the important sources for finding active molecules, however, because the content of the neuropeptide or polypeptide toxin components containing disulfide bonds is low and the stability is poor, no good separation, detection and identification method exists at present, and a method capable of rapidly enriching and detecting the neuropeptides and toxins containing disulfide bonds is urgently needed.
The existing methods for enriching sulfur-containing compounds mainly comprise: covalent chromatography, immobilized metal ion chromatography, molecularly imprinted polymers, gold nanomaterials, and the like. However, the method generally has the problems of complicated operation and long time consumption; some methods can only enrich one sulfur-containing compound, and have poor universality and small application range.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to overcome the defects of the prior art and provides core-shell Fe capable of selectively enriching and separating sulfur-containing compounds3O4a/PDA/Au composite magnetic nano materialThe preparation method can realize the rapid enrichment and separation of the sulfur-containing compounds from the complex sample matrix.
The invention can solve the technical problems that: the process of selectively separating the sulfur-containing compounds by the single gold nanoparticles mainly comprises the steps of adsorbing the sulfur-containing compounds, washing to remove non-adsorbed non-sulfur compounds, and eluting the sulfur-containing compounds adsorbed on the surfaces of the gold nanoparticles. Because the gold nanoparticles are small in particle size, gold nanoparticles adsorbing sulfhydryl compounds are separated from a sample solution through high-speed centrifugal operation, multiple centrifugal separation operations are involved in the whole enrichment and separation process, the operation is complex, the time is consumed, and the used gold nanoparticles cannot be recycled. Fe3O4The nano particles can be quickly separated from the solution under an external magnetic field due to the magnetism, and have the characteristics of simple and quick operation. At present, the research on preparing the gold-magnetic composite nano particles mostly directly coats gold nano particles on the surfaces of the magnetic nano particles, and the effect is not ideal. Polydopamine (PDA has important function in the fields of biomedicine, biosensing and the like due to good hydrophilicity, biocompatibility and electrochemical characteristics3O4The modification has the advantages of simple, convenient, rapid and green method; preparation of Fe at present3O4The method for synthesizing nanometer material of PDA/Au mainly uses PDA to reduce HAuCl in situ4(chloroauric acid) in Fe3O4The PDA which is not coated by the gold nano-particles is easy to react with amino groups, sulfydryl groups and other groups in target molecules to form covalent bonds, so that the enrichment and separation of sulfur-containing compounds are not facilitated. The invention adopts a seed growth method and uses NH2Repeated reduction of HAuCl with OH & HCl as reducing agent4(chloroauric acid), forming a gold nano-layer on the surface of the PDA, and finally preparing the core-shell Fe3O4the/PDA/Au material has good hydrophilicity, dispersibility and biocompatibility, and can be applied to selective enrichment separation of sulfur-containing compounds in the fields of biology, chemistry, medicine and the like.
The technical scheme is as follows: the invention relates to core-shell Fe capable of selectively enriching and separating sulfur-containing compounds3O4PDA/Au magnetic nano material, which is made of magnetic Fe3O4Taking dopamine as an inner core, forming a PDA layer on the surface of the dopamine by oxidation self-polymerization under alkaline conditions, and reducing HAuCl by THPC4Obtaining gold nanoparticles with small particle size to be adsorbed on Fe3O4PDA surface and as crystal nucleus, using NH2Reduction of HAuCl by OH & HCl4Forming a complete gold nano-layer, and finally preparing the core-shell type Fe3O4the/PDA/Au composite magnetic nano material.
In particular, core-shell Fe3O4The preparation method of the/PDA/Au magnetic nano material comprises the following steps:
PDA coated Fe3O4Preparation of nanoparticles:
taking Fe with the same mass3O4(100nm) nanoparticles and dopamine hydrochloride, adding a Tris-HCl buffer solution with the pH value of 8.5, controlling the concentration of dopamine to be 2mg/mL, stirring at room temperature for 4h, separating under an external magnetic field, washing with pure water for multiple times, and obtaining the PDA-coated Fe3O4Dispersing the nanoparticles in pure water;
b. preparing a gold nano seed solution: dissolving 24 μ L of 80% THPC (tetrakis (hydroxymethyl) phosphonium chloride) in 2mL of pure water, adding 90mL of pure water and 1mL of 1mol/L sodium hydroxide solution, stirring for 5min, adding 1% HAuCl4After stirring was continued for 5min, the reaction was terminated.
c. Fe loaded with gold nano-seeds3O4Preparation of PDA: taking Fe in the a3O4Adding the gold nanoparticle seed solution in the step b into the PDA suspension, adjusting the volume ratio of the two solutions to be less than or equal to 1:5, adjusting the pH of the reaction solution to be 2-3 by using 1% citric acid solution, stirring at room temperature for 0.5-1 h, carrying out magnetic separation, and washing by using pure water. Gold nano-seeds with electronegativity are adsorbed on Fe through electrostatic action3O4Surface of PDA to obtain Fe3O4the/PDA-Au seeds were dispersed in pure water.
d. Preparation of three-layer core-shell structure Fe3O4PDA/Au nano material: (1) take 10mLFe3O4The PDA-Au seed dispersion is added with 50mL of pure water and stirred1 min; (2) 0.2mL of 1% HAuCl was added4Continuously stirring for 1 min; (3) 0.375mL of 0.2mol/L NH was added2OH & HCl, and reacting for 10 min; (4) 0.25mL of 1% HAuCl was added to the reaction mixture4And 0.125mL of 0.2mol/L NH2And (4) repeating the step (4) for 6-9 times by using OH & HCl. Magnetic separation and washing are carried out to finally obtain core-shell type Fe3O4the/PDA/Au composite magnetic nano material.
The specific adsorption steps are as follows: adding Fe into sample solution containing sulfur compound3O4PDA/Au magnetic nanomaterial, Fe3O4the/PDA/Au magnetic nano material is fully contacted with the sample solution, after the sample solution is gently shaken for a certain time, under the action of an external magnetic field, the supernatant without sulfur-containing compounds is removed, dithiothreitol, mercaptoethanol or thioglycolic acid are added to absorb Fe3O4And eluting sulfur-containing compounds on the surface of the/PDA/Au nano material.
Through a large number of experimental researches, the invention discovers that Chinese patent 201110190350.5 utilizes PDA to reduce HAuCl in situ4The (chloroauric acid) forms gold nano on the surface of the PDA, the gold nano particles are distributed in star points on the surface of the PDA, a finished gold nano layer cannot be formed, and the PDA is easy to be irreversibly combined with groups such as sulfydryl, amino and the like in a target molecule because a large amount of PDA is not coated by the gold nano particles, so that the PDA cannot be applied to enrichment and separation of sulfur-containing compounds. Chinese patent 201710343036.3 SiO2Coated with Fe3O4Modified at Fe3O4/SiO2The amino and the sulfhydryl on the gold nanoparticle are combined, the required reagents are various, the preparation process is complex, the time consumption is long (16-24h), and a complete gold nanoparticle layer cannot be formed, so that the enrichment of subsequent sulfur-containing compounds is not facilitated. Chinese patent 201210236102.4 SiO2Coated with Fe3O4Preparing gold nano seed solution by using THPC as reducing agent, and reducing HAuCl for multiple times by using formaldehyde as reducing agent4Forming gold nanolayers, although shortening SiO2Coated with Fe3O4But using formaldehyde to reduce HAuCl4When the gold nano-layer is formed, the three-step reduction condition is complex and takes long time (the coating time of the gold nano-particles in the whole process is as long as 47h), the preparation process of the material is complex,and compared with PDA, SiO2Has no better biocompatibility. The invention firstly adsorbs small-grain-size gold nanoparticles on the surface of PDA as the seeds for gold growth, and NH is used2And (3) reducing OH & HCl serving as a reducing agent on the surface of the alloy for multiple times to obtain a complete gold nano layer, and finally preparing the Fe3O4/PDA/Au magnetic nano material with the core-shell structure. Therefore, compared with the comparison document, the invention has the advantages that:
1. THPC (tetrakis (hydroxymethyl) phosphonium chloride) is used as a reducing agent to prepare a gold nano seed solution, and the diameter of gold nano particles is about 2nm, so that the gold nano particles are more uniformly distributed on the surface of the PDA;
2. using NH2Repeated reduction of HAuCl by OH & HCl4The gold nano thin layer can be formed on the surface of the PDA, and the prepared material is more uniform and is more beneficial to the adsorption of the sulfur-containing compound, so that the rapid enrichment and separation of the sulfur-containing compound are realized;
3. the preparation time is short, and the preparation of the material can be completed within 4-6 h.
Firstly, preparing amino modified core-shell structure Fe3O4PDA nano-particles; simultaneous reduction of HAuCl with THPC4Obtaining the gold nano-particles with negative electricity with small particle size (2 nm) and easy to be adsorbed to the Fe modified by the amino3O4PDA surface, and can be used as seed for next step to form gold nano layer; then the above Fe3O4Mixing the PDA and the gold nanoparticle suspension evenly and stirring to obtain the assembled gold-magnetic composite particle Fe3O4PDA/Au; finally using Fe3O4Gold nanoparticles adsorbed on surface of PDA/Au and reduced by THPC are used as crystal nucleus, and NH is used2OH & HCl is subjected to multi-step reduction on the surface of the alloy to obtain a layer of complete gold shell, so that three-layer core-shell structure gold magnetic nanoparticles Fe are prepared3O4/PDA/Au。
Three-layer core-shell structure gold magnetic nanoparticle Fe prepared by the method3O4the/PDA/Au has the particle size of about 120nm, and has good dispersibility, biocompatibility and magnetic responsiveness.
Has the advantages that:
(1) direct in-situ reduction of HAuCl using PDA4The invention utilizes self-assembly technology and seed growth method to prepare core-shell type Fe3O4The PDA/Au magnetic nano material adopts HAuCl which is reduced for many times in the process of preparing the gold nano layer4In the mode, the prepared nanoparticles have uniform particle size distribution, and complete and uniform gold nanolayers can be generated on the surfaces of the nanoparticles.
(2) Compared with the method for enriching and separating the sulfur-containing compound from the nonmagnetic gold nanoparticles, the addition of the magnetic nano material saves the operations of repeated centrifugation and filtration, so that the whole preparation process is simple and quick, and the method is particularly suitable for enriching and separating the easily oxidized sulfhydryl compound.
Drawings
FIG. 1 is Fe3O4Particles; fe3O4PDA particles; core-shell structure gold magnetic nanoparticle Fe3O4Infrared spectrogram of/PDA/Au.
FIG. 2 shows core-shell structure of magnetic nanoparticles Fe3O4Transmission electron microscopy images of/PDA/Au and intermediates in the reduction process. FIG. 2a shows Fe3O4Transmission electron micrographs of the particles; FIG. 2b is Fe3O4Transmission electron micrograph of PDA particles; FIG. 2c shows core-shell particles Fe3O4Transmission electron micrograph of/PDA/Au.
FIG. 3 shows core-shell structure of magnetic nanoparticles Fe3O4X-ray photoelectron spectroscopy analysis of PDA/Au and intermediates in the reduction process. FIG. 3a is Fe3O4X-ray photoelectron spectroscopy analysis chart of PDA particle; FIG. 3b is Fe3O4X-ray photoelectron spectroscopy analysis chart of/PDA/Au particle; FIG. 3c is Fe3O4X-ray photoelectron spectroscopy of Au in PDA/Au particles.
FIG. 4 is a high performance liquid chromatogram before and after enrichment of a sample solution. FIG. 4a shows no addition of Fe3O4High performance liquid chromatogram of sample solution of PDA/Au material; FIG. 4b shows the addition of Fe3O4High performance liquid chromatogram of the sample solution after enrichment of PDA/Au material.
FIG. 5 is a graph showing the comparison of the content of natural products sulforaphane and sulforaphane before and after enrichment.
Detailed Description
Example 1 core-Shell Fe3O4Preparation of/PDA/Au magnetic nano material
Core-shell type Fe3O4The preparation process of the/PDA/Au magnetic nano material comprises the following steps:
preparation of PDA-coated Fe3O4Nanoparticles; ② THPC is used for reducing HAuCl4Obtaining gold nanoparticles with small particle size; thirdly, gold nano particles are adsorbed on Fe through electrostatic action and PDA viscosity3O4The surface of PDA; fourthly, by NH2Multiple reduction of HAuCl by OH & HCl4Forming a complete gold nano-layer, and preparing Fe with a three-layer core-shell structure3O4PDA/Au nano material.
The specific experimental procedures are as follows:
a. preparation of PDA-coated Fe3O4Nanoparticle: weighing Fe3O4Adding 100mg of (100nm) nanoparticles and dopamine hydrochloride into 50ml of Tris-HCl buffer solution with pH of 8.5, stirring at room temperature for 4h, separating under an external magnetic field, washing with pure water for multiple times, and preparing the PDA-coated Fe3O4The nanoparticles were dispersed in 10mL of pure water. 584cm in the infrared spectrum of FIG. 1-1The absorption peak is Fe3O4Characteristic absorption peaks of surface Fe-O bonds; 3426cm-1Is the O-H stretching vibration peak influenced by hydrogen bonds. Fe3O4PDA and Fe3O4Nanoparticle to nanoparticle ratio, Fe3O4The absorption peak of Fe-O bond in PDA is red-shifted. At 2961 and 2848cm-1The absorption peak appears at the position is the C-H stretching vibration on the PDA; at 1618cm-1、1509cm-1、1445cm-1And 1261cm-1The absorption peaks at (A) are respectively the vibration absorption of C-O, N-H on PDA, C-C on the aromatic ring and C-OH on the aromatic ring. Description of PDA in Fe3O4Forming a PDA layer on the surface, and adding Fe3O4And is coated in the coating layer.
b. Preparing a gold nano seed solution: dissolving 24 μ L of 80% THPC in 2mL of pureIn water, 90mL of pure water and 1mL of 1mol/L sodium hydroxide solution were added, and after stirring for 5min, 1% HAuCl was added4After stirring was continued for 5min, the reaction was terminated.
c. Fe loaded with gold nano-seeds3O4Preparation of PDA: 5mL of Fe in a3O4Adding 25mL of the gold nanoparticle seed solution in the step b into the PDA suspension, adjusting the pH of the reaction solution to 2-3 by using a 1% citric acid solution, stirring at room temperature for 1h, carrying out magnetic separation, and washing by using pure water. Gold nano-seeds with electronegativity are adsorbed on Fe through electrostatic interaction and PDA viscosity3O4PDA surface to obtain Fe3O4the/PDA-Au seeds were dispersed in 20mL of pure water.
d. Preparation of three-layer core-shell structure Fe3O4PDA/Au nano material: (1) 10mL of Fe was taken3O4Adding 50mL of pure water into the PDA-Au seed dispersion liquid, and stirring for 1 min; (2) 0.2mL of 1% HAuCl was added4Continuously stirring for 1 min; (3) 0.375mL of 0.2mol/L NH was added2OH & HCl, and reacting for 10 min; (4) 0.25mL of 1% HAuCl was added to the reaction mixture4And 0.125mL of 0.2mol/L NH2OH. HCl, the above step (4) was repeated 9 times. Magnetic separation and washing are carried out to finally obtain core-shell type Fe3O4the/PDA/Au composite magnetic nano material. FIGS. 2a to c are each Fe3O4、Fe3O4PDA and Fe3O4Transmission electron micrograph of/PDA/Au. PDA is easy to generate oxidative autopolymerization in alkaline solution and forms a PDA layer on the surface of the substrate, as shown in a in figure 2, Fe3O4The primary particle size is about 100nm, and the PDA forms Fe after self-gathering on the surface3O4PDA is a ball type structure, where PDA thickness is about 20nm (FIG. 2 b); NH (NH)2Reduction of HAuCl by OH & HCl4The formed gold particles continuously grow and wrap on the surface of the THPC-Au seed to finally form a gold nano shell layer (figure 2 c). Comparison of Fe with XPS3O4/PDA、Fe3O4the/PDA/Au surface element composition is verified. As shown in FIGS. 3 a-b, the elements C, O, N, Au are present in Fe3O4PDA/Au surface. The binding of Fe2p3 at 710.20eV was not detected, further demonstrating Fe in the composite3O4The coating is completely coated by PDA, and the result is consistent with that of the transmission electron microscope image; as shown in FIG. 3c, there are two peaks at 83.0 and 86.6eV, ascribed to Au0The excitation of Au 4f electrons proves that the gold nanoparticles are coated on the surface of the PDA. The results of transmission electron microscope, X-ray photoelectron spectroscopy analysis and infrared analysis show that the invention combines the experimental scheme of adopting self-assembly technology and seed growth method to obtain the three-layer structure gold magnetic nanoparticle Fe coated with complete gold shell3O4/PDA/Au。
e. And (3) performance verification: the specific adsorption steps are as follows: 50mg of Fe prepared above was immersed in 10mL of a mixed sample solution (containing cysteine, glutathione, tyrosine and tryptophan)3O4PDA/Au magnetic nanomaterial, Fe3O4the/PDA/Au magnetic nano material is fully contacted with the sample solution, after the sample solution is gently shaken for a certain time, under the external magnetic field, the supernatant fluid without sulfhydryl compound is removed, dithiothreitol is added to absorb Fe3O4And eluting the sulfhydryl compound on the surface of the/PDA/Au nano material. FIGS. 4a and 4b show the addition of Fe3O4High performance liquid chromatogram of mixed sample before and after enrichment of PDA/Au, as shown in figure, cysteine and glutathione which are sulfhydryl-containing compounds in the solution are adsorbed by the material, while the signal intensity of tyrosine and tryptophan which are non-sulfhydryl-containing compounds is kept unchanged, which proves that Fe3O4the/PDA/Au material has selective adsorption to only sulfhydryl compounds. (chromatographic peak assignment: 1. tyrosine; 2. cysteine; 3. tryptophan; 4. glutathione).
Example 2 enrichment of thiol-rich Polypeptides in proteomic Studies
(1) Extraction and enzymolysis of mouse brain tissue protein
Taking about 5mg of mouse hypothalamus tissue, adding 40 μ l of protein extract (50 mM Tris-HCl solution of 4% SDS), ultrasonically extracting until the sample is dissolved, adding 10mM DTT, incubating for 30min, adding 55mM Iodoacetamide (IAA), incubating for 30min in the dark, adding 10mM DTT, adding 220 μ l of 80% acetone, precipitating at 4 ℃ overnight, centrifuging at 16000rpm, discarding the supernatant, adding 220 μ l of 80% acetone, standing for 1 hour, discarding the supernatant, standing at room temperature for 12min until the acetone is completely volatilized. Adding protein lysate (Tris-HCl buffer solution containing 8M urea) for full dissolution, diluting with Tris-HCl buffer solution until the concentration of urea is lower than 1M, adding trypsin according to the mass percent of 1%, performing enzymolysis for 12 hours at 37 ℃ to obtain enzymolysis solution, then loading the enzymolysis solution on a Seppak C18 column, washing with trifluoroacetic acid, eluting with 80% acetonitrile solution containing 0.2% trifluoroacetic acid, and collecting eluent.
(2) Enrichment of sulfur-containing peptide fragments
Immersing 50mg of Fe in the eluent obtained in the step (1)3O4PDA/Au magnetic nanomaterial (prepared according to inventive example 1), let Fe3O4the/PDA/Au magnetic nano material is fully contacted with the sample solution, after the sample solution is gently shaken for a certain time, under the external magnetic field, the supernatant fluid without sulfhydryl compound is removed, dithiothreitol is added to absorb Fe3O4And eluting the sulfhydryl compound on the surface of the/PDA/Au nano material to obtain the sulfur-containing polypeptide part.
(3) Nano LC-MS/MS high-throughput identification of peptide fragment sequence
Injecting the sulfur-containing polypeptide obtained in the step (2) into a Danan U3000 NanoRSLC nanoliter liquid phase system, wherein a chromatographic column is 5 mu m Reprosil C18AQ (75 mu m multiplied by 150mm), the loading amount is 1-2 mu L, the flow rate is 200-400 nL/min, a mobile phase A (acetonitrile/formic acid/water is 2/0.2/98, v/v/v), a mobile phase B (acetonitrile/formic acid/water is 80/0.2/20, v/v/v), and 2-30% B is eluted for 150min in a linear gradient manner;
and using a Thermo Q-active Orbitrap mass spectrometer for peptide fragment analysis, wherein the spray voltage is 2.5kV, and the temperature of an ion transmission capillary is 200 ℃; the primary full-scanning range of the mass spectrum is 300-2000 m/z, and the separation width is 3 Da; the tandem mass spectrometry adopts a secondary mass spectrometry scanning mode depending on primary mass spectrometry data, sequentially selects 5 ions with the highest ion intensity in the primary mass spectrometry to carry out Collision Induced Dissociation (CID) secondary tandem mass spectrometry, and adopts Xcalibur software to carry out data acquisition;
performing library searching, identifying and analyzing on the secondary mass spectrum data by using PEAKS 8.5 software, selecting a corresponding protein database, and setting retrieval parameters as follows: error of 10ppm for precursor ion; the error of the daughter ion is 1 Da; allowing 2 sites to be cut by mistake, wherein the false positive rate is less than or equal to 1 percent; selecting pancreatin (Trypsin) in a digestion mode, wherein the number of unique peptide segments is more than or equal to 2; other parameters are default parameters, the score obtained under the search conditions has significance, and P <0.05 is determined as a valid identification result; and identifying and determining the amino acid sequences of all peptide fragments in each sample enzymolysis liquid.
The result identified 6623 peptide fragments altogether, in which the number of peptide fragments containing Cys (cysteine) was 4650, and the thiol-containing peptide fragments were enriched to 70%. Of 6623 peptides identified, 1107 peptides containing Met (methionine, another sulfur-containing amino acid) accounted for about 17%, and thus Fe by the present invention3O4The PDA/Au magnetic nano material can better finish the enrichment of the peptide section containing sulfur, especially the peptide section containing sulfydryl.
EXAMPLE 3 enrichment of thiol-containing peptide fragments in extracts of keratinous-derived animal drugs
(1) Weighing cornu Bubali, slicing, pulverizing to obtain powder, adding 20 times of water, decocting for 3 times, each time for 4 hr, mixing extractive solutions, and concentrating to appropriate concentration to obtain cornu Bubali water extract.
(2) Enrichment of sulfur-containing peptide fragments
Immersing 50mg Fe in the buffalo horn water extract obtained in the step (1)3O4PDA/Au magnetic nanomaterial (prepared according to inventive example 1), let Fe3O4the/PDA/Au magnetic nano material is fully contacted with the sample solution, after the sample solution is gently shaken for a certain time, under the external magnetic field, the supernatant fluid without sulfhydryl compound is removed, dithiothreitol or mercaptoethanol is added to absorb the adsorbed Fe3O4And eluting the sulfhydryl compound on the surface of the/PDA/Au nano material to obtain the buffalo horn sulfhydryl-containing polypeptide part.
(3) Nano LC-MS/MS high-throughput identification of peptide fragment sequence
Desalting the buffalo horn sulfhydryl-containing polypeptide part obtained in the step (2) by using SeppakC18, drying, re-dissolving by using an initial mobile phase, injecting into a Dyan U3000 NanoRSLC nanoliter liquid phase system, wherein a chromatographic column is 5 mu m Reprosil C18AQ (75 mu m multiplied by 150mm), the sample loading amount is 1-2 mu L, the flow rate is 200-400 nL/min, the mobile phase A (acetonitrile/formic acid/water is 2/0.2/98, v/v/v), the mobile phase B (acetonitrile/formic acid/water is 80/0.2/20, v/v/v), and 2-30% B is eluted for 150min in a linear gradient manner;
and using a Thermo Q-active Orbitrap mass spectrometer for peptide fragment analysis, wherein the spray voltage is 2.5kV, and the temperature of an ion transmission capillary is 200 ℃; the primary full-scanning range of the mass spectrum is 300-2000 m/z, and the separation width is 3 Da; the tandem mass spectrometry adopts a secondary mass spectrometry scanning mode depending on primary mass spectrometry data, sequentially selects 5 ions with the highest ion intensity in the primary mass spectrometry to carry out Collision Induced Dissociation (CID) secondary tandem mass spectrometry, and adopts Xcalibur software to carry out data acquisition;
performing library searching, identification and analysis on the secondary mass spectrum data by using PEAKS 8.5 software, selecting a bovine protein database, and setting retrieval parameters as follows: error of 10ppm for precursor ion; the error of the daughter ion is 1 Da; allowing 2 sites to be cut by mistake, wherein the false positive rate is less than or equal to 1 percent; selecting Non-enzyme digestion (Non enzyme) in an enzyme digestion mode, wherein the number of unique peptide segments is more than or equal to 2; other parameters are default parameters, the score obtained under the search conditions has significance, and P <0.05 is determined as a valid identification result; and identifying and determining the amino acid sequences of all peptide fragments in each sample enzymolysis liquid.
The result identifies 1019 peptide segments, wherein the number of the peptide segments containing Cys (cysteine) is 737, and the enrichment of the peptide segments containing sulfydryl reaches 72%. Some of the thiol-containing peptides identified contain 4-5 Cys, such as TITPCISSPCAPAAPCTPCVPR, AQASCCRPSYCGQSCCR, RPVCCDPCSLQEGCCR, SCQAVVCRPCCW. It can be seen that by Fe3O4The PDA/Au magnetic nano material can better complete the enrichment of peptide sections containing sulfydryl in the extract liquid of the keratinous traditional Chinese medicines.
EXAMPLE 4 enrichment of Natural products sulforaphane and glucoraphanin
(1) Sulforaphane and sulforaphane glycoside are abundant in cruciferous plants and are common antioxidants. The broccoli contains sulforaphane and sulforaphane glycoside, 500g of broccoli is cut into small blocks, 500ml of water is added for decoction and extraction for 30min, the mixture is concentrated to a proper amount, and the volume is determined to be 100ml, so that broccoli extracting solution is obtained.
(2) Enrichment of sulforaphane and sulforaphane glycoside
Immersing 50mg of Fe into 100ml of broccoli extract obtained in the step (1)3O4PDA/Au magnetic nanomaterial (prepared according to inventive example 1), let Fe3O4The PDA/Au magnetic nano material is fully contacted with the sample solution, after the sample solution is gently shaken for a certain time, under the external magnetic field, the supernatant fluid without sulfhydryl compound is removed, cysteine is added to adsorb the Fe3O4Eluting sulfhydryl compound on the surface of the/PDA/Au nano material to obtain enriched broccoli extract, and fixing the volume to 100ml to obtain broccoli enriched liquid.
(3) Determination of sulforaphane and sulforaphane glycoside
Chromatographic conditions are as follows: wondasil C18 column (4.6 mm. times.250 mm,5 μm); mobile phase: performing gradient elution with methanol-water for 0-20 min, wherein the concentration of methanol is increased from 1% to 30%; the detection wavelength is 254 nm; the flow rate is 1.0 ml/min; the column temperature is 30 ℃; the number of theoretical plates is not less than 5000 based on sulforaphane.
And respectively carrying out HPLC analysis on the broccoli extracting solution and the broccoli enriched solution, and determining the contents of the sulforaphen and the sulforaphane glycoside in the two samples by adopting a standard curve method. As shown in FIG. 5 before and after enrichment, the contents of sulforaphane and sulforaphane glycoside in the broccoli extract before enrichment were determined as follows: 58ng/ml and 1806ng/ml, while the contents of sulforaphane and sulforaphane glycoside in the enriched broccoli solution after enrichment are 1289ng/ml and 23 mug/ml respectively, and the sulforaphane and sulforaphane glycoside are enriched by 22.4 times and 12.7 times respectively, which shows that Fe is used for enriching the radish3O4The PDA/Au magnetic nano material can better complete the enrichment of two sulfur-containing compounds, namely sulforaphane and sulforaphane glycoside.
Claims (7)
1. The nanometer material capable of selectively enriching and separating sulfur-containing compounds is characterized in that the nanometer material is core-shell Fe3O4the/PDA/Au composite magnetic nano material.
2. The preparation method of the nano material capable of selectively enriching and separating the sulfur-containing compounds is characterized by comprising the following steps of:
mixing Fe3O4Uniformly mixing PDA nanoparticles with gold nano solution with small particle size to obtain Fe3O4The gold nanoparticles adsorbed on the surface of the PDA are taken as crystal nuclei, and NH is utilized2Multiple reduction of HAuCl by OH & HCl4Obtaining a complete gold nano-layer, thereby preparing the core-shell type Fe3O4PDA/Au nano-material;
3. the method for preparing the nano-material capable of selectively enriching and separating the sulfur-containing compounds according to claim 2, is characterized by comprising the following steps:
PDA coated Fe3O4Preparation of nanoparticles:
taking Fe with the same mass3O4Adding nanoparticles and dopamine hydrochloride into Tris-HCl buffer solution with the pH value of 8.5, controlling the concentration of the dopamine hydrochloride to be 2mg/mL, stirring for 4 hours at room temperature, separating under an external magnetic field, washing with pure water for multiple times, and preparing the PDA-coated Fe3O4Nano particles, and dispersing the nano particles in pure water;
b. preparing a gold nano seed solution:
dissolving 24 μ L of 80% THPC in 2mL of pure water, adding 90mL of pure water and 1mL of 1mol/L sodium hydroxide solution, stirring for 5min, adding 1% HAuCl4After stirring for 5min, the reaction is ended;
c. fe loaded with gold nano-seeds3O4Preparation of PDA:
taking the PDA coated Fe prepared in the step a3O4Adding the prepared gold nano-seed solution into the nano-particle suspension, adjusting the pH of the solution to 2-3 by using 1% citric acid solution, stirring at room temperature for 0.5-1 h, carrying out magnetic separation, washing with pure water, and adsorbing the gold nano-seeds with electronegativity on Fe through electrostatic action3O4Surface of PDA to obtain Fe3O4Dispersing the/PDA-Au seeds in pure water;
d. preparation of core-shell-structured Fe3O4PDA/Au nano material:
(1) take 10mLFe3O4Adding 50mL of pure water into the PDA-Au seed dispersion liquid, and stirring for 1 min;
(2) 0.2mL of 1% HAuCl was added4Continuously stirring for 1 min;
(3) 0.375mL of 0.2mol/L NH was added2OH & HCl, and reacting for 10 min;
(4) 0.25mL of 1% HAuCL was added to the reaction mixture4And 0.125mL of 0.2mol/L NH2OH·HCl；
(5) Repeating the step (4) for 6-9 times, and then carrying out magnetic separation and washing to obtain the core-shell Fe3O4the/PDA/Au composite magnetic nano material.
4. A method for enriching and separating sulfur-containing compounds is characterized by comprising the following steps:
adding a sample solution containing a sulfur compound to the core-shell type Fe3O4After the PDA/Au composite magnetic nano material is slightly shaken for a certain time, under the action of an external magnetic field, supernatant fluid without sulfur compounds is removed; or slowly flowing a sample solution containing sulfur-containing compound through the core-shell Fe3O4In a test tube of/PDA/Au composite magnetic nano material; then adding dithiothreitol, mercaptoethanol, thioglycolic acid or cysteine to adsorb the core-shell Fe3O4And eluting sulfur-containing compounds on the surface of the/PDA/Au composite magnetic nano material.
5. The method for enriching and separating sulfur-containing compounds according to claim 4, wherein said sulfur-containing compounds comprise thiol-containing compounds such as cysteine, homocysteine, glutathione, etc.; disulfide bond-containing compounds such as cystine and homocystine; methionine bond-containing compounds such as methionine and S-adenosylmethionine; thiol-containing proteins and peptides, such as cysteine-containing proteins and peptides; peptides and toxins containing disulfide bonds, including neuropeptide, conotoxin, scorpion toxin, and spider toxin; the sulfur-containing natural products include alkaloids, terpenoids, saponins, and alkane natural products.
6. The process for the enriched separation of sulfur-containing compounds according to claim 4, wherein the cysteine, cysteine-containing proteins or cysteine-containing peptides are eluted with dithiothreitol; the peptide and toxin components containing disulfide bonds are washed and removed by thioglycollic acid or cysteine; the sulfur-containing natural product is eluted with dithiothreitol, mercaptoethanol, thioglycolic acid or cysteine.
7. The nanomaterial for selectively enriching and separating sulfur-containing compounds according to claim 1, characterized in that the core-shell type Fe3O4the/PDA/Au composite magnetic nano material is used for the enrichment research of proteins and polypeptides containing sulfydryl in proteomes and polypeptide groups, or the enrichment and detection of sulfydryl-containing components in biological samples such as cysteine, cystine, glutathione, methionine and the like in medical examination, or the enrichment and separation research of peptide toxins containing disulfide bonds in animal-derived toxins, or the enrichment research of sulfur-containing compounds in traditional Chinese medicines and natural products.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113058571A (en) * | 2021-03-23 | 2021-07-02 | 南昌大学 | Preparation method of bifunctional polymer adsorbent and application of bifunctional polymer adsorbent in gold recovery |
| CN114471715A (en) * | 2022-03-02 | 2022-05-13 | 烟台大学 | Preparation method and application of polydopamine-loaded nano-gold catalyst |
| CN116535658A (en) * | 2023-04-14 | 2023-08-04 | 广东新宝堂生物科技有限公司 | Magnetic material for enriching flavonoid compounds in dried orange peel enzyme, preparation method thereof and application thereof in analysis |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101893634A (en) * | 2009-05-20 | 2010-11-24 | 中国科学院生物物理研究所 | Specific detection method of protein or polypeptide cysteine sulfydryl modification and application thereof |
| US20120237605A1 (en) * | 2011-03-15 | 2012-09-20 | Messersmith Phillip B | Multifunctional Metal Nanoparticles Having A Polydopamine-Based Surface and Methods of Making and Using the Same |
| CN102861921A (en) * | 2011-07-08 | 2013-01-09 | 西南民族大学 | Preparation method of core-shell magnetic/gold nano particles |
| CN104162414A (en) * | 2014-08-01 | 2014-11-26 | 曲阜师范大学 | Vacant shell type poly-dopamine adsorbed nanogold composite material as well as preparation and application thereof |
| CN104258909A (en) * | 2014-08-01 | 2015-01-07 | 曲阜师范大学 | Fe3O4-poly-dopamine-Au nano-composite material as well as preparation method and application thereof |
| CN109369912A (en) * | 2018-11-05 | 2019-02-22 | 南方科技大学 | In the method that poly-dopamine nanoparticle surface density is controllably grafted DNA |
-
2019
- 2019-09-19 CN CN201910886669.8A patent/CN112516972B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101893634A (en) * | 2009-05-20 | 2010-11-24 | 中国科学院生物物理研究所 | Specific detection method of protein or polypeptide cysteine sulfydryl modification and application thereof |
| US20120237605A1 (en) * | 2011-03-15 | 2012-09-20 | Messersmith Phillip B | Multifunctional Metal Nanoparticles Having A Polydopamine-Based Surface and Methods of Making and Using the Same |
| CN102861921A (en) * | 2011-07-08 | 2013-01-09 | 西南民族大学 | Preparation method of core-shell magnetic/gold nano particles |
| CN104162414A (en) * | 2014-08-01 | 2014-11-26 | 曲阜师范大学 | Vacant shell type poly-dopamine adsorbed nanogold composite material as well as preparation and application thereof |
| CN104258909A (en) * | 2014-08-01 | 2015-01-07 | 曲阜师范大学 | Fe3O4-poly-dopamine-Au nano-composite material as well as preparation method and application thereof |
| CN109369912A (en) * | 2018-11-05 | 2019-02-22 | 南方科技大学 | In the method that poly-dopamine nanoparticle surface density is controllably grafted DNA |
Non-Patent Citations (2)
| Title |
|---|
| CHENXU WANG ET AL.: "Robust Nanoparticle-DNA Conjugates Based on Mussel-Inspired Polydopamine Coating for Cell Imaging and Tailored Self-Assembly", 《BIOCONJUGATE CHEMISTRY》 * |
| 顾觉奋, 中国医药科技出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113058571A (en) * | 2021-03-23 | 2021-07-02 | 南昌大学 | Preparation method of bifunctional polymer adsorbent and application of bifunctional polymer adsorbent in gold recovery |
| CN114471715A (en) * | 2022-03-02 | 2022-05-13 | 烟台大学 | Preparation method and application of polydopamine-loaded nano-gold catalyst |
| CN114471715B (en) * | 2022-03-02 | 2023-10-24 | 烟台大学 | Preparation method and application of polydopamine loaded nano-gold catalyst |
| CN116535658A (en) * | 2023-04-14 | 2023-08-04 | 广东新宝堂生物科技有限公司 | Magnetic material for enriching flavonoid compounds in dried orange peel enzyme, preparation method thereof and application thereof in analysis |
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