AU2018201824B2

AU2018201824B2 - Method of making colloidal metal nanoparticles

Info

Publication number: AU2018201824B2
Application number: AU2018201824A
Authority: AU
Inventors: Ta-Wei Chang; Chun-Lun Chiu; Kuei-Sheng Fan; Jim-Min Fang; Lin Lu; Chen-Hsiang Wang; Cheng-ding WANG
Original assignee: Tripod Technology Corp
Current assignee: Tripod Technology Corp
Priority date: 2017-06-19
Filing date: 2018-03-14
Publication date: 2019-11-14
Anticipated expiration: 2038-03-14
Also published as: AU2018201824A1; JP2019002067A; DE102017116090A1; DE102017116090B4; TWI648099B; JP6461262B2; TW201904658A

Abstract

Provided is a method of making colloidal metal nanoparticles. The method includes the steps of: mixing a metal aqueous solution and a reducing agent to form a mixture solution in a reaction tank; heating the mixture solution and 5 undergoing a reduction reaction to produce a composition containing metal nanoparticles, residues and gas, wherein the amount of the residues is less than 20 % by volume of the mixture solution, and guiding the gas out of the reaction tank; dispersing the metal nanoparticles with a medium to obtain colloidal metal nanoparticles; wherein the metal aqueous solution contains metal ions 10 and anions, and the metal ions comprise gold ions, silver ions, copper ions, zinc ions, nickel ions, palladium ions, cobalt ions, iron ions, titanium ions, cadmium ions, platinum ions, aluminum ions, lead ions, manganese ions, chromium ions, molybdenum ions, vanadium ions, or tungsten ions, and the anions comprise halide ions or nitrate ions. By separating the reduction reaction step and the 15 dispersion step, the method of making colloidal metal nanoparticles is simple, safe, time-effective, cost-effective, and has the advantage of high yield. MIXING A METAL AQUEOUS SOLUTION AND A REDUCING AGENT TO FORM A MIXTURE SOLUTION IN A REACTION TANK HEATING THE MIXTURE SOLUTION AND UNDERGOING A REDUCTION REACTION TO PRODUCE A COMPOSITION CONTAINING METAL NANOPARTICLES, RESIDUES AND GAS; AND GUIDING SAID GAS OUT OF THE REACTION TANK DISPERSING THE METAL NANOPARTICLES WITH A MEDIUM TO OBTAIN COLLOIDAL METAL NANOPARTICLES

Description

METHOD OF MAKING COLLOIDAL METAL NANOPARTICLES

1. Field of the Invention

The present disclosure relates to a method of making colloidal metal nanoparticles.

2. Description of the Related Art(s)

Colloidal metal nanoparticles exhibit the optical, electromagnetic and chemical properties distinct from those of bulk materials because of their small size effect, surface effect, and quantum size effect. Therefore, metal nanoparticles have a wide range of applications in materials science, information science, catalysis and life sciences. In recent years, scientists have been actively developing various methods to produce metal nanoparticles. The methods for making metal nanoparticles can be divided into three major categories:(l) laser ablation method, which uses high-energy laser for continuous irradiation on a metal bulk material; (2) metal vapor synthesis, which condenses vaporized gaseous metal atoms under controlled conditions for aggregation; and (3) chemical reduction method, which reduces metal ions to metal nanoparticles in solutions. At present, the chemical reduction method is most commonly and conveniently utilized in preparation of metal nanoparticles. The reduction reactions can be performed in water or organic solvents.

Having large surface area, metal nanoparticles display high physical and chemical activities for easy oxidation and agglomeration. Thus, a variety of modifiers or capping agents are often introduced in the chemical preparation of metal nanoparticles for control of the particle size, shape, distribution, dispersion and stability. The technology for making metal nanoparticle colloids has a great influence on the stability and quality of the product. However, adding those stabilizers make the producing method more complicated.

To overcome the problem, some methods have been provided.

US 8,048,193 discloses a method for producing gold colloid. The method includes a nucleation step of forming nuclear colloidal particles by adding a citrate reducing agent to a first gold salt solution; and a growth step of growing nuclear colloid, which is necessarily performed at least once, by adding a second gold salt and an ascorbate reducing agent to the solution of the nuclear colloidal particles. Although said method could produce gold colloids having a targeted particle size and a uniform spherical shape, the method could not be performed as desired unless using the particular reagents and restricted steps. In order to obtain larger particle size of gold colloids, the number of the growth step should be increased. As a result, said method in the prior art still is not performed conveniently and efficiently.

US 20120046482 discloses a method for synthesizing gold nanoparticles. A gold ion containing solution and a carboxylic acid including at least two carboxyl groups are mixed to form a mixture and reacted at a reaction temperature of about 20°C to about 60°C. Although said method could produce gold nanoparticles simply, the method could not be performed as desired unless using the particular reagents. Said method limits the reacting step at a relatively low reaction temperature, so said reacting step needs more time to complete, and some reduction agents could not be applied in this method. As a result, said method in the prior art still is not performed conveniently and efficiently.

In view that the conventional methods fail to produce colloidal metal nanoparticles conveniently and efficiently, the instant disclosure is directed to a method and the system for making colloidal metal nanoparticles that may provide a high yield product in a manner which may be relatively simple, safe, time-effective, cost-effective, and environment-friendly.

More particularly, the instant disclosure provides a method of making colloidal metal nanoparticles including steps (a) to (c). In step (a), a metal aqueous solution is mixed with a reducing agent to form a mixture solution in a reaction tank. In step (b), the mixture solution is heated and undergoes a reduction reaction to produce a composition containing metal nanoparticles, residues and gas, wherein the amount of the residues less than 20 % by volume of the mixture solution, and guiding said gas out of the reaction tank, wherein the temperature of the heating ranges from 70°C to 150°C. In step (c), the metal nanoparticles are dispersed with a medium to obtain colloidal metal nanoparticles.

The metal aqueous solution contains metal ions, and the metal ions comprise gold ions, silver ions, copper ions, zinc ions, nickel ions, palladium ions, cobalt ions, iron ions, titanium ions, cadmium ions, platinum ions, aluminum ions, lead ions, manganese ions, chromium ions, molybdenum ions, vanadium ions, or tungsten ions.

The metal aqueous solution also contains anions, and the anions comprise halide ions or nitrate ions.

By means of heating the mixture solution and guiding the gas produced from the reduction reaction out of the reaction tank, the reduction reaction can react completely, and then the yield can be improved. Also, limiting the volume of the mixture solution in the reduction reaction can enhance collision probability of reactant atoms, so that the reaction rate can accelerate. As water medium is vaporized during formation of metal nanoparticles, the reduction reaction step and the dispersion step will not proceed at the same time. Moreover, the reducing agents and dispersing agents have a wide range of choices and will not be restricted. Accordingly, the present method is useful to simplify the production of colloidal metal nanoparticles.

In some cases, the metal ions are from HAuC1₄, AgNO₃, Cu(NO₃)₂, CuCl₂, ZnCl₂, NiCl₂, PdCl₂, CoCl₂, FeCl₂, FeCl₃, TiCl₂, TiCl₃, or TiCl₄.

In addition, trapping the gas produced from the reduction reaction with water can collect a large amount of aqueous acids to be recovered for reuse, which can reduce the acid wastes.

In certain embodiments, HAuC1₄ is used for making gold nanoparticles, and the gaseous HC1 generated from the reduction reaction is trapped with water to make hydrochloric acid for recovery.

In certain embodiments, AgNO₃ is used for making silver nanoparticles, and the gaseous NO₂ generated from the reduction reaction is trapped with water to make nitric acid for recovery.

The reaction temperature affects the reaction rate of making metal nanoparticles. Without proper temperature control, the reaction will proceed unevenly and may generate bubble to affect the quality of metal nanoparticles.

In accordance with the instant disclosure, the temperature of the heating in the step (b) ranges from 50°C to 150°C. Preferably, the temperature of the heating in the step (b) ranges from 70°C to 130°C.

In accordance with the instant disclosure, the temperature of the dispersing in the step (c) ranges from 20°C to 100°C. Preferably, the temperature of the dispersing in the step (c) ranges from 50°C to 80°C.

In accordance with the instant disclosure, the reducing agent may comprise at least one ester.

In some cases, said ester is selected from the group of a carboxylate ester, a cyclic ester, a polymeric ester, and combinations thereof.

Preferably, said carboxylate ester is represented by the formula (I), ΗΟ^^ΛΖ^Οχ^χ+ι (I), wherein R is H or CH₃, and x is an integer ranging from to 16.

Preferably, said cyclic ester is represented by the formula (II), (II), wherein the ring contains one oxygen atom and 4 to 6 carbon atoms, and G is H, CH₃ or C₂H₅.

Preferably, said polymeric ester is represented by the formula (III),

Γ Ο Ί

HO.

H

R ⁿ (III), wherein R is H or CH₃, and n is an integer ranging from 2 to 1400.

Preferably, said combination of ester reducing agents is a methyl lactate and an ethyl lactate, a methyl lactate and γ-butyrolactone, or an ethyl lactate and γ-butyrolactone.

In accordance with the instant disclosure, the reducing agent may comprise a citric acid, a lactic acid, a glycolic acid, an ascorbic acid, an oxalic acid, a tartaric acid, a 1,4-butanediol, a glycerol, a polyethylene glycol), a hydroquinone, an acetaldehyde, a glucose, a cellulose, a carboxymethyl cellulose, a cyclodextrin, a chitin, a chitosan, or combinations thereof.

In some cases, the reducing agent may comprise a combination of at least one ester and at least one non-ester reducing agent.

Preferably, said combination of at least one ester and at least one non-ester reducing agent is a methyl lactate in combination with a lactic acid, a citric acid, 1,4-butanediol, or a poly(ethylene glycol).

Preferably, said combination of at least one ester and at least one non-ester reducing agent is an ethyl lactate in combination with a lactic acid, a citric acid, 1,4-butanediol, or a poly(ethylene glycol).

Preferably, said combination of at least one ester and at least one non-ester reducing agent is a γ-butyrolactone in combination with a lactic acid.

In some cases, the molar concentration of the metal aqueous solution ranges from 0.1 M to 3.0 M. Preferably, the molar concentration of the metal aqueous solution ranges from 0.1 M to 1.0 M. More preferably, the molar concentration of the metal aqueous solution is 0.2 M.

In some cases, the reducing agent comprises a polymeric ester or a first reagent. When the reducing agent comprises the first reagent, the first reagent is selected from the group of a carboxylate ester, a cyclic ester, a citric acid, a lactic acid, a glycolic acid, an ascorbic acid, an oxalic acid, a tartaric acid, a

1,4-butanediol, a glycerol, a hydroquinone, an acetaldehyde, a glucose, a chitin, and combinations thereof, a molar ratio of the first reagent relative to the metal ions ranges from 1 to 40. Preferably, the molar ratio of the first reagent relative to the metal ions ranges from 1 to 8. More preferably, the molar ratio of the first reagent relative to the metal ions is 4.

In some cases, the ester is a polymeric ester, and the weight of said polymeric ester ranges from 30 mg to 150 mg.

In accordance with the instant disclosure, the reaction time ranges from 5 minutes to 80 minutes depending on the kind of the reducing agent and the molar concentration of reactants. Preferably, the reaction time ranges from 7 minutes to 15 minutes.

The reduction rate of gold ion can be tuned by a combined use of reducing agents to give gold nanoparticles in various sizes.

In accordance with the instant disclosure, the medium in step (c) for dispersing the metal nanoparticles may be water or an aqueous solution of dispersing agent.

In some cases, said aqueous solution of dispersing agents may be an aqueous citric acid, an aqueous lactic acid, an aqueous poly(lactic acid), an aqueous sodium hydroxide, an aqueous hexadecylamine, an aqueous oleylamine, an aqueous tetraoctylammonium bromide (TOAB), an aqueous dodecanethiol, an aqueous polyethylene oxide), an aqueous polyvinylpyrrolidone (PVP), or combinations thereof.

The molar concentration of the dispersing agents ranges from 0.001 M to 0.1 M.

Preferably, the molar concentration of the dispersing agents ranges from 0.01 M to 0.05 M.

In some cases, the molar ratio of the dispersing agent to metal nanoparticles ranges from 10 to 100.

Preferably, the molar ratio of the dispersing agent to metal nanoparticles ranges from 15 to 30.

In accordance with the instant disclosure, all the water used in the aqueous solution is a distilled water. More preferably, the water is a deionized water.

It is beneficial to use only organic reducing agents and organic dispersing agents in water without the involvement of inorganic cations such as Na⁺ or K⁺ to process the reduction reaction in step (b) and dispersing in step (c). Accordingly, said colloidal metal nanoparticles will attain a good stability without the interference of inorganic cations.

In accordance with the instant disclosure, the process of the reduction reaction is monitored by the infrared (IR) spectral analysis on-site.

The region from about 1500 cm'¹ to 500 cm'¹ of the IR spectrum contains a very complicated series of absorptions. These are mainly due to all manners of bond vibrations within the molecule. This region is called the fingerprint region. The importance of the fingerprint region is that each different substance produces a different pattern of troughs in this part of the spectrum. Therefore, the pattern of troughs different from the original mixture solution means the reaction proceeds, and when the pattern of troughs is not changed apparently, the reaction is complete.

In accordance with the instant disclosure, the size of colloidal metal nanoparticles may be characterized by the ultraviolet-visible (UV-Vis) spectral absorption because the wavelength at absorption maximum (Imax) of the colloidal metal nanoparticles in UV-Vis spectrum has its own specific range.

In some cases, the Xmax of colloidal gold nanoparticles ranges from 515 nm to 572 nm.

In some cases, the Xmax of colloidal silver nanoparticles ranges from 370 nm to 420 nm.

Besides the kinds of the metal, the size of colloidal metal nanoparticles also affects the Xmax in UV-Vis spectrum. An increase in wavelength Xmax correlates to an increase in the size of the nanoparticle. For example, the colloidal gold nanoparticles with Xmax at 525 nm correspond to their average size of 26 nm ± 1 nm; the colloidal gold nanoparticles with Xmax at 530 nm correspond to their average size of 30 nm ± 1 nm.

In accordance with the instant disclosure, the size of metal nanoparticles also can be characterized by transmission electron microscopy (TEM) imaging.

In accordance with the instant disclosure, the colloidal metal nanoparticles show high zeta potential, which is a key indicator of the stability of colloidal dispersion.

Other objectives, advantages and novel features of the disclosure will become more apparent from the following detailed description when taken in conjunction with the accompanying drawings.

IN THE DRAWINGS

Fig. 1 is a schematic flow diagram illustrating a method for making colloidal metal nanoparticles in accordance with the present invention;

Fig. 2 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 2;

Fig. 3 A is a FT-IR spectra of colloidal gold nanoparticles obtained in Example 2 (as shown in thin line) and product of heating methyl lactate with aqueous HC1 at 130°C for 12 minutes (as shown in thick line);

Fig. 3B is a FT-IR spectra of product of heating methyl lactate with aqueous HC1 at 130°C for 12 minutes (as shown in thin line) and starting material of methyl lactate (as shown in thick line);

Fig. 4 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 5;

Fig. 5 is TEM image of gold nanoparticles (mean diameter of 22 nm-23 nm) obtained in Example 5;

Fig. 6 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 15;

Fig. 7A is a FT-IR spectra of colloidal gold nanoparticles obtained in Example 15(as shown in thin line) and product of heating ethyl lactate with aqueous HC1 at 130°C for 12 minutes (as shown in thick line);

Fig. 7B is a FT-IR spectra of product of heating ethyl lactate with aqueous HC1 at I30°C for 12 minutes(as shown in thin line) and starting material of ethyl lactate (as shown in thick line);

Fig. 8 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 23;

Fig. 9 is TEM image of gold nanoparticles (mean diameter of 33 nm-34 nm) obtained in Example 23;

Fig. 10 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 27;

Fig. 11A is a FT-IR spectra of colloidal gold nanoparticles obtained in Example 27(as shown in thick line) and product of heating γ-butyrolactone with aqueous HC1 at 130°C for 30 minutes (as shown in thin line);

Fig. 1 IB is a FT-IR spectra of product of heating γ-butyrolactone with aqueous HC1 at 130°C for 30 minutes (as shown in thin line) and starting material of γ-butyrolactone (as shown in thick line);

Fig. 12 is TEM image of gold nanoparticles (mean diameter of 27 nm-28 nm) obtained in Example 27;

Fig. 13 is UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 33;

Fig. 14A is a FT-IR spectra of colloidal gold nanoparticles obtained in Example 33 (as shown in thick line) and product of heating poly(lactic acid) with aqueous HC1 at 130°C for 30 minutes (as shown in thin line);

Fig. 14B is FT-IR spectrum of starting material of poly(lactic acid);

Fig. 15 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 35;

Fig. 16 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 37;

Figs. 17A is a FT-IR spectra of colloidal gold nanoparticles obtained in Example 37 (as shown in thin line) and product of heating lactic acid with aqueous HC1 at 130°C for 12 minutes (as shown in thick line);

Figs. 17B is a FT-IR spectra of product of heating lactic acid with aqueous HC1 at 130°C for 12 minutes (as shown in thin line) and starting material of lactic acid(as shown in thick line);

Fig. 18 is a zeta potential diagram of colloidal gold nanoparticles obtained in Example 37;

Fig. 19 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 38;

Fig. 20 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 44;

Fig. 21 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 45;

Fig. 22 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 46;

Fig. 23 is a UV-Vis spectrum of colloidal gold nanoparticles obtained in Example 47;

Fig. 24 is TEM image of gold nanoparticles (mean diameter of 38 nm-39 nm) obtained in Example 47;

Fig. 25 is a UV-Vis spectrum of colloidal silver nanoparticles obtained in Example 58;

Fig. 26 is TEM image of colloidal silver nanoparticles (mean diameter of 10 nm-11 nm) obtained in Example 58;

Fig. 27 is a UV-Vis spectrum of colloidal silver nanoparticles obtained in Example 59;

Fig. 28 is a UV-Vis spectrum of colloidal silver nanoparticles obtained in Example 60;

Fig. 29 is a UV-Vis spectrum of colloidal silver nanoparticles obtained in Example 61;

Fig. 30 is TEM image of colloidal palladium nanoparticles (mean diameter of 9 nm to 10 nm) obtained in Example 65; and

Fig. 31 is TEM image of colloidal zinc nanoparticles (mean diameter of 26 nm to27 nm) obtained in Example 71.

Hereinafter, one skilled in the arts can easily realize the advantages and effects of the instant disclosure from the following examples. Therefore, it should be understood that the descriptions proposed herein are just preferable examples for the purpose of illustrations only, not intended to limit the scope of the disclosure. Various modifications and variations could be made in order to practice or apply the instant disclosure without departing from the spirit and scope of the disclosure.

Process of Making Colloidal Metal Nanoparticles

In the following examples, infrared (IR) spectra were recorded on Agilent Technologies Cary630 Fourier transform (FT)-IR spectrometer. Ultraviolet-visible (UV-Vis) spectra were measured on Agilent Technologies Cary60 UV-Vis spectrophotometer. Transmission electron microscopy (TEM) images were recorded on Hitachi H-7100 microscope. All the reagents were reagent grade and used as purchase without further purification. Tetrachloroauric acid (HAuC1₄, 0.2 M aqueous solution) and zinc powder were purchased from Acros Organics (New Jersey, USA). Silver nitrate (AgNO₃, 0.1 M aqueous solution) was purchased from Merck & Co. (New Jersey, USA). Palladium chloride (PdCB, containing 59.4% Pd) was purchased from Uni

Region Bio-Tech (Taipei, Taiwan). Ultra-pure water was purchased from Hao Feng Biotech Co. (Taipei, Taiwan).

Example 1 Synthesis of colloidal gold nanoparticles using methyl lactate as reducing and dispersing agent

Hereinafter, the process of making colloidal metal nanoparticles was conducted by using the method as shown in Fig. 1.

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (22.3 mg, 0.21 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 30 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of methyl lactate (200 mg, 1.9 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 550 nm with OD = 0.365.

Example 2 Synthesis of colloidal gold nanoparticles using methyl lactate as reducing agent and citric acid as dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (20.6 mg, 0.20 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 15 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 545 nm with OD = 3.896 as in Fig.2, (measured at 50% dilution). In addition, formation of colloidal gold nanoparticles was confirmed by the FT-IR spectrum as shown in Fig.3 A.

Example 3 Synthesis of colloidal gold nanoparticles using methyl lactate as reducing agent and citric acid as dispersing agent without heating

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (20.8 mg, 0.20 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was stirred at room temperature for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at /.max = 545 nm with OD = 1.784.

Example 4 Synthesis of colloidal gold nanoparticles using methyl lactate and 1,4-butanediol as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (10.7 mg, 0.10 mmol) and

1,4-butanediol (10.2 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 10.5 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg,

1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 1.258.

Example 5 Synthesis of colloidal gold nanoparticles using methyl lactate and citric acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (14.7 mg, 0.14 mmol) and citric acid (20.9 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.274 as in Fig.4, (measured at 50% dilution). The mean diameter of gold nanoparticles was 22 nm to 23 nm as shown by TEM image in Fig.5.

Example 6 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG800 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (12.1 mg, 0.12 mmol) and PEG800 (80.4 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 15.5 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg,

1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 3.172.

Example 7 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG 1000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (11.2 mg, 0.12 mmol) and PEG1000 (106.3 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 3.118.

Example 8 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG4000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (12.1 mg, 0.12 mmol) and PEG4000 (402.7 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 3.412.

Example 9 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG8000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (11.6 mg, 0.11 mmol) and PEG8000 (808.2 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at /max = 535 nm with OD = 2.952.

Example 10 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG 10000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (11 mg, 0.11 mmol) and PEG10000 (1.003 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at /..max = 530 nm with OD = 3.028.

Example 11 Synthesis of colloidal gold nanoparticles using methyl lactate and PEG 11000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (11.9 mg, 0.11 mmol) and PEG11000 (1.104 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 3.548.

Example 12 Synthesis of colloidal gold nanoparticles using methyl lactate and lactic acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (11.8 mg, 0.11 mmol) and lactic acid (13.1 mg, 0.15 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 15 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 3.49.

Example 13 Synthesis of colloidal gold nanoparticles using methyl lactate and ethyl lactate as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and methyl lactate (10.4 mg, 0.10 mmol) and ethyl lactate (13.1 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 8.5 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 1.844.

Example 14 Synthesis of colloidal gold nanoparticles using ethyl lactate as reducing and dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (26.7 mg, 0.23 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of ethyl lactate (200 mg, 1.7 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 555 nm with OD = 0.397.

Example 15 Synthesis of colloidal gold nanoparticles using ethyl lactate as reducing agent and citric acid as dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mF of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (25.9 mg, 0.22 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 10 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD =1.641 as in Fig.6. In addition, formation of colloidal gold nanoparticles was confirmed by the FT-IR spectrum as shown in Fig.7A.

Example 16 Synthesis of colloidal gold nanoparticles using ethyl lactate as reducing agent and citric acid as dispersing agent without heating

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (23.6 mg, 0.20 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an

Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 30°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.148.

Example 17 Synthesis of colloidal gold nanoparticles using ethyl lactate and lactic acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (11.6 mg, 0.10 mmol) and lactic acid (11.3 mg, 0.13 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 1.996.

Example 18 Synthesis of colloidal gold nanoparticles using ethyl lactate and 1,4-butanediol as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (11.5 mg, 0.10 mmol) and 1,4-butanediol (10.6 mg, 0.12 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 1.017.

Example 19 Synthesis of colloidal gold nanoparticles using ethyl lactate and citric acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (17.2 mg, 0.15 mmol) and citric acid (20.9 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.324.

Example 20 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG800 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (12.2 mg, 0.10 mmol) and PEG800 (81.4 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 20 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 3.802.

Example 21 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG 1000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (14.0 mg, 0.12 mmol) and PEG1000 (103 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at /..max = 525 nm with OD = 3.086.

Example 22 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG4000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (12.9 mg, 0.11 mmol) and PEG4000 (402.8 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 3.012.

Example 23 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG8000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mF of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (11 mg, 0.09 mmol) and PEG8000 (807.5 mg) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 30 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mF water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 3.1 as in Fig.8. The mean diameter of gold nanoparticles was 33 nm to 34 nm as shown by TEM image in Fig.9.

Example 24 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG 10000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (12.1 mg, 0.10 mmol) and PEG10000 (1.005 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 2.882.

Example 25 Synthesis of colloidal gold nanoparticles using ethyl lactate and PEG 11000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and ethyl lactate (12.5 mg, 0.11 mmol) and PEG11000 (1.07 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 540 nm with OD = 2.996.

Example 26 Synthesis of colloidal gold nanoparticles using γ-butyrolactone as reducing agent and sodium hydroxide as dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (18.4 mg, 0.21 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of NaOH (46.5 mg,

1.2 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 565 nm with OD = 0.134.

Example 27 Synthesis of colloidal gold nanoparticles using γ-butyrolactone as reducing agent and citric acid as dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (19.3 mg, 0.22 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 2.866 as in Fig. 10 (measured at 50% dilution). The mean diameter of gold nanoparticles was 27 nm to 28 nm as shown by TEM image in Fig. 12. In addition, formation of colloidal gold nanoparticles was confirmed by the FT-IR spectrum as shown in Fig. 11 A.

Example 28 Synthesis of colloidal gold nanoparticles using γ-butyrolactone as reducing agent and citric acid as dispersing agent with heating at 50°C

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (17.2 mg, 0.20 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 50°C for 10 minutes to

2018201824 14 Mar 2018 obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.29.

Example 29 Synthesis of colloidal gold nanoparticles using γ-butyrolactone and lactic acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (10.2 mg, 0.12 mmol) and lactic acid (11.3 mg, 0.13 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 18 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 1.582.

Example 30 Synthesis of colloidal gold nanoparticles using γ-butyrolactone and methyl lactate as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (11.9 mg, 0.14 mmol) and methyl lactate (11.3 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 17 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 0.463.

Example 31 Synthesis of colloidal gold nanoparticles using γ-butyrolactone and methyl lactate as combined reducing agent with a short reaction time

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 1.496

Example 32 Synthesis of colloidal gold nanoparticles using γ-butyrolactone and ethyl lactate as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and γ-butyrolactone (11.1 mg, 0.13 mmol) and ethyl lactate (12.2 mg, 0.10 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 1.416.

Example 33 Synthesis of colloidal gold nanoparticles using poly(lactic acid) as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and poly(lactic acid) (PLA) (90.6 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.426 as Fig 13 (measured at 50% dilution). In addition, formation of gold nanoparticles was confirmed by FT-IR spectrum as shown in Fig. 14A.

Example 34 Synthesis of colloidal gold nanoparticles using poly(lactic acid) as reducing agent with heating at 60 °C

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and poly(lactic acid) (90.8 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 60°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was less than 20% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.444.

Example 35 Synthesis of colloidal gold nanoparticles using glycolic acid as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and glycolic acid (19.1 mg, 0.25 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 5.226 as in Fig. 15, (measured at 50% dilution).

Example 36 Synthesis of colloidal gold nanoparticles using lactic acid as reducing and dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mF of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (20.4 mg, 0.23 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 15 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mF water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of lactic acid (220 mg, 2.4 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 0.897.

Example 37 Synthesis of colloidal gold nanoparticles using lactic acid as reducing agent and citric acid as dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mF of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (21.3 mg, 0.24 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 9 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.468 as in Fig. 16 (measured at 50% dilution). In addition, Formation of gold nanoparticles was confirmed by FT-IR spectrum as shown in Fig.l7A. Moreover, the zeta potential of colloidal gold nanoparticles was -44.86 mV shown as in Fig. 18.

Example 38 Synthesis of colloidal gold nanoparticles using citric acid as reducing

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and citric acid (40.3 mg, 0.21 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), 50 mL of pure water was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.412 as in Fig. 19.

Example 39 Larger scale synthesis of colloidal gold nanoparticles using citric acid as reducing

First, in step (a), Tetrachloroauric acid (10 mL of 0.2 M aqueous solution, 2 mmol) and citric acid (1.6 g, 83.4 mmol) were added via an inlet port into a double-necked flat-bottomed 2 L reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 14 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), 2 L of pure water was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.433.

Example 40 Synthesis of colloidal gold nanoparticles using citric acid as reducing agent and glycerol as dispersing agent

First, in step (a), Tetrachloroauric acid (0.5 mL of 0.2 M aqueous solution, 0.1 mmol) and citric acid (80.8 mg, 0.42 mmol) were added via an inlet port into a double-necked flat-bottomed 150 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (100 mL) of glycerol (400 mg,

4.3 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.472.

Example 41 Synthesis of colloidal gold nanoparticles using citric acid as reducing agent and PEG as dispersing agent

First, in step (a), Tetrachloroauric acid (0.5 mL of 0.2 M aqueous solution, 0.1 mmol) and citric acid (81.3 mg, 0.42 mmol) were added via an inlet port into a double-necked flat-bottomed 150 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (100 mL) of PEG800 (400 mg) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.13.

Example 42 Synthesis of colloidal gold nanoparticles using citric acid as reducing agent with heating at 150°C

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and citric acid (40.2 mg, 0.21 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 2 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), 50 mL of pure water was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 1.961.

Example 43 Synthesis of colloidal gold nanoparticles using citric acid as reducing agent with heating at 70°C

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and citric acid (40.8 mg, 0.21 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 70°C for 40 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was less than 20% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), 50 mL of pure water was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.492.

Example 44 Synthesis of colloidal gold nanoparticles using cellulose as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and cellulose (40.8 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 4.17 as in Fig. 20, (measured at 50% dilution).

Example 45 Synthesis of colloidal gold nanoparticles using carboxymethyl cellulose as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and carboxymethyl cellulose (40 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 3.528 as in Fig. 21 (measured at 50% dilution).

Example 46 Synthesis of colloidal gold nanoparticles using chitin as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and chitin (41.6 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 567 nm with OD = 0.216 as in Fig. 22.

Example 47 Synthesis of colloidal gold nanoparticles using chitosan as reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and chitosan (81.6 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 538 nm with OD = 0.162 as in Fig. 23. The mean diameter of gold nanoparticles was estimated to be 38 nm to 39 nm as shown by TEM image in Fig.24; however, the shape and size of said gold nanoparticles were not homogeneous.

Example 48 Synthesis of colloidal gold nanoparticles using poly(vinylpyrrolidone) as reducing agent and dispersing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and poly(vinylpyrrolidone) (PVP, 48.5 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 80 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of PVP (200 mg) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 60°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 2.76.

Example 49 Synthesis of colloidal gold nanoparticles using lactic acid and 1,4-butanediol as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mF of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.8 mg, 0.13 mmol) and 1,4-butanediol (10.8 mg, 0.12 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mF water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of citric acid (200 mg,

1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 530 nm with OD = 1.254.

Example 50 Synthesis of colloidal gold nanoparticles using lactic acid and citric acid as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (15.5 mg, 0.17 mmol) and citric acid (20.3 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and was mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.872.

Example 51 Synthesis of colloidal gold nanoparticles using lactic acid and PEG800 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.3 mg, 0.13 mmol) and PEG800 (80.9 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 130°C for 17.5 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing gold nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.906.

Example 52 Synthesis of colloidal gold nanoparticles using lactic acid and PEG 1000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.2 mg, 0.12 mmol) and PEG1000 (101.9 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.996.

Example 53 Synthesis of colloidal gold nanoparticles using lactic acid and PEG4000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.1 mg, 0.12 mmol) and PEG4000 (400.2 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 525 nm with OD = 2.836.

Example 54 Synthesis of colloidal gold nanoparticles using lactic acid and PEG8000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (10.5 mg, 0.12 mmol) and PEG8000 (802.3 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 3.166.

Example 55 Synthesis of colloidal gold nanoparticles using lactic acid and PEG 10000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.7 mg, 0.13 mmol) and PEG10000 (1.042 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 3.12.

Example 56 Synthesis of colloidal gold nanoparticles using lactic acid and PEG 11000 as combined reducing agent

First, in step (a), Tetrachloroauric acid (0.25 mL of 0.2 M aqueous solution, 0.05 mmol) and lactic acid (11.7 mg, 0.13 mmol) and PEG11000 (1.109 g) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (200 mg, 1.0 mmol) was used as a medium to disperse the gold nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal gold nanoparticles, which showed the UV-Vis absorption band at Xmax = 535 nm with OD = 3.282.

Example 57 Synthesis of colloidal silver nanoparticles using methyl lactate as reducing agent

First, in step (a), silver nitrate (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and methyl lactate (24.5 mg, 0.24 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing silver nanoparticles, residues and NO₂ gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, NO₂ gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) andNaOH (46.5 mg, 1.12 mmol) was used as a medium to disperse the silver nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 15 minutes to obtain colloidal silver nanoparticles, which showed the UV-Vis absorption band at Xmax = 390 nm with OD = 2.433.

Example 58 Synthesis of colloidal silver nanoparticles using methyl lactate and citric acid as combined reducing agent

First, in step (a), silver nitrate (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.9 mg, 0.24 mmol) and methyl lactate (11.4 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) andNaOH (46.5 mg, 1.12 mmol) was used as a medium to disperse the silver nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 15 minutes to obtain colloidal silver nanoparticles, which showed the UV-Vis absorption band at Xmax = 390 nm with OD = 2.882 as in Fig.25 (measured at 50% dilution). The mean diameter of colloidal silver nanoparticles was 10 nm tol 1 nm as shown by TEM image in Fig.26.

Example 59 Synthesis of colloidal silver nanoparticles using ethyl lactate and citric acid as combined reducing agent

First, in step (a), silver nitrate (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.9 mg, 0.24 mmol) and ethyl lactate (11.7 mg, 0.10 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg,

0.17 mmol) andNaOH (46.5 mg, 1.12 mmol) was used as a medium to disperse the silver nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 15 minutes to obtain colloidal silver nanoparticles, which showed the UV-Vis absorption band at Xmax = 390 nm with OD = 2.812 as in Fig.27 (measured at 50% dilution).

Example 60 Synthesis of colloidal silver nanoparticles using lactic acid and citric acid as combined reducing agent

First, in step (a), silver nitrate (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.4 mg, 0.24 mmol) and lactic acid (10.3 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) andNaOH (46.5 mg, 1.12 mmol) was used as a medium to disperse the silver nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 15 minutes to obtain colloidal silver nanoparticles, which showed the UV-Vis absorption band at Xmax = 390 nm with OD = 2.798 as in Fig.28 (measured at 50% dilution).

Example 61 Synthesis of colloidal silver nanoparticles using citric acid as reducing and dispersing agent

First, in step (a), silver nitrate (0.1 mF of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.1 mg, 0.23 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing silver nanoparticles, residues and NO₂ gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, NO₂ gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mF water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of citric acid (33.6 mg, 0.17 mmol) andNaOH (46.5 mg, 1.12 mmol) was used as a medium to disperse the silver nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 10 minutes to obtain colloidal silver nanoparticles, which showed the UV-Vis absorption band at Xmax = 390 nm with OD = 2.602 as shown in Fig.29 (measured at 50% dilution).

Example 62 Synthesis of colloidal palladium nanoparticles using ethyl lactate as reducing agent

First, in step (a), palladium chloride (0.1 mF of 0.1 M aqueous solution,

0.01 mmol) and ethyl lactate (27.8 mg, 0.24 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing palladium nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) was used as a medium to disperse the palladium nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal palladium nanoparticles.

Example 63 Synthesis of colloidal palladium nanoparticles using γ-butyrolactone as reducing agent

First, in step (a), palladium chloride (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and γ-butyrolactone (20.3 mg, 0.24 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Example 64 Synthesis of colloidal palladium nanoparticles using methyl lactate and citric acid as combined reducing agent

First, in step (a), palladium chloride (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.7 mg, 0.24 mmol) and methyl lactate (11.7 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg,

0.17 mmol) was used as a medium to disperse the palladium nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal palladium nanoparticles.

Example 65 Synthesis of colloidal palladium nanoparticles using ethyl lactate and citric acid as combined reducing agent

First, in step (a), palladium chloride (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.4 mg, 0.24 mmol) and ethyl lactate (11.9 mg, 0.10 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) was used as a medium to disperse the palladium nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal palladium nanoparticles. The mean diameter of colloidal palladium nanoparticles was 9 nm to 10 nm as shown by TEM image in Fig.30.

Example 66 Synthesis of colloidal palladium nanoparticles using lactic acid and citric acid as combined reducing agent

First, in step (a), palladium chloride (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.7 mg, 0.24 mmol) and lactic acid (10.8 mg, 0.12 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Example 67 Synthesis of colloidal palladium nanoparticles using citric acid as reducing and dispersing agent

First, in step (a), palladium chloride (0.1 mL of 0.1 M aqueous solution, 0.01 mmol) and citric acid (45.2 mg, 0.23 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Example 68 Synthesis of colloidal zinc nanoparticles using poly(lactic acid) as reducing agent

First, in step (a), zinc chloride (0.1 mL of 2 M aqueous solution, 0.2 mmol) and poly(lactic acid) (106.5 mg) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing zinc nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an Erlenmeyer flask for collection.

Finally, in step (c), an aqueous solution (50 mF) of citric acid (33.6 mg, 0.17 mmol) was used as a medium to disperse the zinc nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal zinc nanoparticles.

Example 69 Synthesis of colloidal zinc nanoparticles using methyl lactate and citric acid as combined reducing agent

First, in step (a), zinc chloride (0.1 mF of 2 M aqueous solution, 0.2 mmol) and citric acid (45.9 mg, 0.24 mmol) and methyl lactate (10.4 mg, 0.10 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mF reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing zinc nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mF water in an Erlenmeyer flask for collection.

Example 70 Synthesis of colloidal zinc nanoparticles using ethyl lactate and citric acid as combined reducing agent

First, in step (a), zinc chloride (0.1 mL of 2 M aqueous solution, 0.2 mmol) and citric acid (45.9 mg, 0.24 mmol) and ethyl lactate (11.4 mg, 0.10 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) was used as a medium to disperse the zinc nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal zinc nanoparticles.

Example 71 Synthesis of colloidal zinc nanoparticles using lactic acid and citric acid as combined reducing agent

First, in step (a), zinc chloride (0.1 mL of 2 M aqueous solution, 0.2 mmol) and citric acid (45.9 mg, 0.24 mmol) and lactic acid (10.2 mg, 0.11 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Finally, in step (c), an aqueous solution (50 mL) of citric acid (33.6 mg, 0.17 mmol) was used as a medium to disperse the zinc nanoparticles in the flat-bottomed flask, and said solution was heated at 70°C for 30 minutes to obtain colloidal zinc nanoparticles. The mean diameter of colloidal zinc nanoparticles was 26 nm to 27 nm as shown by TEM image in Fig.31.

Example 72 Synthesis of colloidal zinc nanoparticles using citric acid as reducing and dispersing agent

First, in step (a), zinc chloride (0.1 mL of 2 M aqueous solution, 0.2 mmol) and citric acid (45.9 mg, 0.24 mmol) were added via an inlet port into a double-necked flat-bottomed 100 mL reaction flask and were mixed to form a mixture solution.

Subsequently, in step (b), the flat-bottomed flask was placed on a hot plate and heated at 150°C for 12 minutes to perform a reduction reaction which was monitored by the IR spectrometer. The reduction reaction produced a composition containing zinc nanoparticles, residues and HC1 gas; the amount of the residues was almost 0% by volume of the mixture solution. At the same time, HC1 gas produced from the reduction reaction was through the recovery port attached to the flat-bottomed flask and was trapped with 10 mL water in an

Erlenmeyer flask for collection.

Discussion of the results

Based on the results of Examples 1 to 56, the instant process employs aqueous Tetrachloroauric acid solution as metal source and varies different kinds of reducing agents to form gold nanoparticles, and then uses diverse kinds of medium to disperse said gold nanoparticles to obtain colloidal gold nanoparticles.

From the results of Examples 57 to 61, the instant process employs aqueous silver nitrate solution as metal source and varies different kinds of reducing agents to form silver nanoparticles, and then uses diverse kinds of medium to disperse said silver nanoparticles to obtain colloidal silver nanoparticles.

From the results of Examples 62 to 67, the instant process employs aqueous palladium chloride solution as metal source and varies different kinds of reducing agents to form palladium nanoparticles, and then uses diverse kinds of medium to disperse said palladium nanoparticles to obtain colloidal palladium nanoparticles.

From the results of Examples 68 to 72, the instant process employs aqueous zinc chloride solution as metal source and varies different kinds of reducing agents to form zinc nanoparticles, and then using diverse kinds of

2018201824 14 Mar 2018 medium to disperse said zinc nanoparticles to obtain colloidal zinc nanoparticles.

Further, Examples 1 to 34, Example 57 to 59, Example 62 to 65, and Example 68 to 70 use non-toxic and biocompatible reducing agents of esters including methyl lactate, ethyl lactate, γ-butyrolactone or poly(lactic acid). It is more eco-friendly and suitable to be applied in the present society.

From the comparison results of Examples 1 and 2, selection of different dispersing media for the metal nanoparticles to make colloidal metal nanoparticles in various sizes is determined from the different Xmax. Similarly, from the comparison results of Examples 14 and 15, they also have colloidal metal nanoparticles in different mean sizes. In addition, from the comparison results of Examples 2, 15 and 35, selection of different reducing agents to form the metal nanoparticles in various sizes is determined from the different Xmax. As the method is processed and separated by two steps rather than in one pot reaction, it can have wider range of options to choose suitable reducing agents and dispersing medium. Accordingly, it is more convenient to apply in various industrial and medical applications.

Compared with the conventional process, the concentration of metal ions is relatively high in Examples 1 to 72 because of the low water content in the reduction reaction, and therefore the reaction time can be reduced for making metal nanoparticles within 80 minutes, in most examples even within 20 minutes, and the fastest is even merely 2 minutes. This is a cost-effective process, and faster reaction rate of reduction yields a narrower size distribution of metal nanoparticles. As a result, said metal nanoparticles in homogeneous size distribution do not require further filtration, so the yield can improve.

Unlike the conventional method that involves a hazardous process in adding a solution of metal ion rapidly to a boiling solution of reducing agent, the instant method by heating a pre-mixed solution of metal ions and reducing agents even in a large scale is a much safer manner. Moreover, the instant method proceeds in an easy and efficient manner by just using simple setup without complicate apparatus of reactor or stirring equipment apparatus.

What is more, using organic reducing and dispersing agents in water makes colloidal metal nanoparticles have good quality and stability without the interference of other inorganic cations.

Besides, according to the step (b) in the instant disclosure, heating and guiding the gas out of the reaction tank can convert the anions (e.g., Cl and NO₃ ) to gas (e.g., HC1 and NO₂) that is trapped by water for reuse. As most anions are removed from the colloidal metal nanoparticles, said colloidal metal nanoparticles have high stability and zeta potential without appreciable interference of anions.

Even though numerous characteristics and advantages of the instant disclosure have been set forth in the foregoing description, together with details of the structure and features of the disclosure, the disclosure is illustrative only. Changes may be made in the details, especially in matters of shape, size, and arrangement of parts within the principles of the disclosure to the full extent indicated by the broad general meaning of the terms in which the appended claims are expressed.

2018201824 14 Mar 2018

It will be understood that the term “comprise” and any of its derivatives (eg comprises, comprising) as used in this specification is to be taken to be inclusive of features to which it refers, and is not meant to exclude the presence of any additional features unless otherwise stated or implied.

The reference to any prior art in this specification is not, and should not be taken as, an acknowledgement of any form of suggestion that such prior art forms part of the common general knowledge.

Claims

The claims defining the invention are as follows:

1. A method of making colloidal metal nanoparticles, comprising steps of:

step (a): mixing a metal aqueous solution and a reducing agent to form a mixture solution in a reaction tank;

step (b): heating the mixture solution and undergoing a reduction reaction to produce a composition containing metal nanoparticles, residues and gas, wherein the amount of the residues is less than 20 % by volume of the mixture solution, and guiding said gas out of the reaction tank, wherein the temperature of the heating ranges from 70°C to 150°C; and step (c): dispersing the metal nanoparticles with a medium to obtain colloidal metal nanoparticles;

wherein the metal aqueous solution contains metal ions and anions, and the metal ions comprise gold ions, silver ions, copper ions, zinc ions, nickel ions, palladium ions, cobalt ions, iron ions, titanium ions, cadmium ions, platinum ions, aluminum ions, lead ions, manganese ions, chromium ions, molybdenum ions, vanadium ions, or tungsten ions, and the anions comprise halide ions or nitrate ions, and wherein the molar concentration of the metal ions ranges from 0.1 M to 3.0 M; and wherein the reducing agent comprises a polymeric ester or a first reagent selected from the group of a carboxylate ester, a cyclic ester, a citric acid, a lactic acid, a glycolic acid, an ascorbic acid, an oxalic acid, a tartaric acid, a

1,4-butanediol, a glycerol, a hydroquinone, an acetaldehyde, a glucose, a chitin, and combinations thereof; and when the reducing agent comprises the first reagent, a molar ratio of the first reagent relative to the metal ions ranges from 1 to 40, and when the reducing agent comprises the polymeric ester, the weight of the polymeric ester ranges from 30 mg to 150 mg; and wherein the carboxylate ester is represented by the formula (I),

2018201824 19 Jun 2019 to 16;

HCk/CO₂C_xH_2x+1 (I), wherein R is H or CH3, and x is an integer ranging from 1 wherein the cyclic ester is represented by the formula (II),

G (II), wherein the ring contains one oxygen atom and 4 to 6 carbon atoms, and G is H, CH₃ or C2H5;

the polymeric ester is represented by the formula (III),

HO.

ⁿ (III), wherein R is H or CH3, and n is an integer ranging from 2 to

1400.
2. The method as claimed in claim 1, wherein the metal ions are from HAuCL, AgNO₃, Cu(NO₃)₂, CuCl₂, ZnCl₂, NiCl₂, PdCl₂, CoCl₂, FeCl₂, FeCl₃, TiCl₂, TiCl₃, or TiCl₄.
3. The method as claimed in claim 1, wherein the step (b) of guiding the gas produced from the reduction reaction out of the reaction tank comprises trapping the gas with water in a tank.
4. The method as claimed in claim 1, wherein the temperature of the heating in the step (b) ranges from 70°C to 130°C.
5. The method as claimed in claim 1, wherein the temperature of the dispersing in the step (c) ranges from 20°C to 100°C.
6. The method as claimed in claim 5, wherein the temperature of the dispersing in the step (c) ranges from 50°C to 80°C.
7. The method as claimed in claim 1, wherein the reducing agent is the carboxylate ester, the cyclic ester, the polymeric ester, or combinations thereof.

2018201824 19 Jun 2019
8. The method as claimed in claim 1 or 7, wherein the reducing agent further comprises a poly(ethylene glycol), a cellulose, a carboxymethyl cellulose, a cyclodextrin, a chitosan, or combinations thereof.
9. The method as claimed in claim 1, wherein the medium in step (c) comprises a dispersing agent with an aqueous citric acid, an aqueous lactic acid, an aqueous poly(lactic acid), an aqueous sodium hydroxide, an aqueous hexadecylamine, an aqueous oleylamine, an aqueous tetraoctylammonium bromide (TOAB), an aqueous dodecanethiol, an aqueous poly(ethylene oxide), an aqueous polyvinylpyrrolidone (PVP), or combinations thereof.
10. The method as claimed in claim 9, wherein a molar concentration of the dispersing agent ranges from 0.001 M to 0.1 M.
11. The method as claimed in claim 9, wherein a molar ratio of the dispersing agent relative to the metal nanoparticles ranges from 10 to 100.
12. The method as claimed in claim 11, wherein a molar ratio of the dispersing agent relative to the metal nanoparticles ranges from 15 to 30.