CN115612156A - Ag NPs-BCM substrate, preparation method thereof and application thereof in cancer marker detection - Google Patents
Ag NPs-BCM substrate, preparation method thereof and application thereof in cancer marker detection Download PDFInfo
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Abstract
The invention discloses an Ag NPs-BCM substrate, a preparation method thereof and application thereof in cancer marker detection, wherein the Ag NPs-BCM substrate grows in situ on three-dimensional Bacterial Cellulose Membrane (BCM) nanofibers and uniformly distributes high-density Ag NPs through controllable silver mirror reaction and volume contraction treatment, so that a highly stable and uniform hot spot is formed on a networked SERS substrate; the Ag NPs-BCM substrate is prepared by soaking in ammonia water to hydrolyze a hemiacetal structure of cellulose, then reducing in situ in a silver trifluoroacetate (CF 3 COOAg) solution, and then drying in vacuum. The preparation method is simple and easy to operate, does not need to add an additional reducing agent, and is safer to use CF 3 COOAg; soaking ammonia water is beneficial to hydrolysis of cellulose and combination with nano cellulose of BCM, and a reliable combination site is provided for reducing silver nano particles; the invention uses the Ag NPs-BCM substrate to detect Glutathione (GSH), distinguishes the GSH content of cancer patients and normal people, and has good application prospect in rapid biomedical detection.
Description
Technical Field
The invention belongs to the technical field of analytical chemistry micro-detection, and particularly relates to an Ag NPs-BCM substrate for SERS detection of cancer markers, and a preparation method and application thereof.
Background
Glutathione (GSH) is a tripeptide naturally synthesized in the human cytoplasm, composed of glutamic acid, cysteine and glycine, and widely distributed in various organs of the body. The molecule contains sulfhydryl group, can combine with free radical in vivo, is an important antioxidant and antidote, and plays an important role in maintaining cell biological function. Glutathione content in blood of normal human body is lower, about 26-34mg/100g, while that of cancer patients (gastric cancer, liver cancer, colon cancer) is 2-4 times higher than that of normal human body.
At present, the most common methods for cancer identification in clinical application are routine examination, tumor marker screening, kit or special examination, which is not only expensive, but also limited in every aspect. Application number CN201911405221.6 provides an in vitro kit for colorectal cancer diagnosis, the preparation process of the kit is relatively complicated, the detection sensitivity is relatively low, and there is a defect in trace detection. Patent No. CN111830178A provides a method for detecting glutathione, which uses a liquid chromatograph and a standard curve equation to detect GSH, but although the detection time is shortened, the operation of the process is too complicated, and is not suitable for detecting a large number of samples.
Surface Enhanced Raman Scattering (SERS) is a molecular detection spectroscopy technique that has wide applications in environmental chemistry, drug detection, biomedicine, and single-molecule detection. In recent years, with the improvement of environmental awareness of people, the SERS substrate which is sustainable, pollution-free, simple, economical and efficient becomes a target pursued by researchers. SERS substrates, among which cellulose is representative, exhibit absolute advantages in the trend.
Cellulose is the most abundant organic matter in the world, and has the characteristics of hydrophilicity, air permeability, degradability, low fluorescence background and the like, so that the cellulose becomes a promising SERS substrate material. The cellulose is used as a solid carrier, a reducing agent and a stabilizing agent to play an important role in the design and manufacture of the SERS substrate, and a flexible SERS substrate with novel style, multiple functions and excellent performance is sequentially appeared.
The natural cellulose-bacterial cellulose membrane prepared by microorganisms gradually occupies an important position in a cellulose family due to the advantages of high moisture retention, high purity, high wet strength and the like brought by the unique three-dimensional network structure and the nanometer effect. Because of their ease of synthesis and biocompatibility, BC has great potential in environmental science, biomedicine, and drug delivery applications.
Disclosure of Invention
The invention aims to provide an Ag NPs-BCM substrate, a preparation method thereof and application thereof in cancer marker detection; loading high-density Ag NPs on a three-dimensional bacterial cellulose membrane to prepare a BCM (Ag NPs-BCM) substrate, and analyzing the content of glutathione in different human serum so as to distinguish cancer patients from normal persons; the method has the advantages of simple preparation, safety, easy obtainment, good reproducibility, high sensitivity, long SERS performance period, low cost and no pollution.
The technical scheme of the invention is as follows:
one of the objects of the present invention is to provide a method for preparing an Ag NPs-BCM substrate, comprising the steps of: after the bacterial cellulose membrane is soaked in ammonia water for volume shrinkage treatment, ag NPs with high density are grown in situ on the three-dimensional bacterial cellulose membrane by utilizing silver mirror reaction and are uniformly distributed, and then the Ag NPs-BCM substrate rich in silver nanospheres is prepared.
Further, the preparation method specifically comprises the following steps:
s1, preparation of a three-dimensional bacterial cellulose membrane: preparing a three-dimensional bacterial cellulose membrane by using fermentation liquor;
s2, pretreatment of the three-dimensional bacterial cellulose membrane: sequentially soaking the three-dimensional bacterial cellulose membrane prepared in the step S1 in ultrapure water, absolute ethyl alcohol and ammonia water for pretreatment to form a three-dimensional bacterial cellulose membrane with a hydrogel texture;
s3, substrate preparation: and (3) adding the three-dimensional bacterial cellulose membrane prepared in the step (S2) into the silver salt solution, and reducing the silver salt solution in the three-dimensional bacterial cellulose membrane through silver mirror reaction to prepare the Ag NPs-BCM substrate rich in silver nanospheres.
Further, the preparation of the three-dimensional bacterial cellulose membrane in the step S1 specifically comprises the following operations:
s11, adding 3-10g of fermentation liquor into 500-1000mL of boiling water, and fully stirring the fermentation liquor;
s12, standing the fermentation liquor, separating to obtain a filtrate, adding 20-60g of white granulated sugar into the filtrate, and fully stirring to obtain a sugar fermentation liquor;
and S13, after the sugar fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 100-500mL of fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator to obtain the three-dimensional bacterial cellulose membrane.
Further, the fermentation liquor is green tea fermentation liquor; the fermentation bacteria mother liquor is green tea bacteria mother liquor; the temperature of the constant temperature incubator is 30-35 ℃.
Further, the three-dimensional bacterial cellulose membrane pretreatment in the step S2 specifically comprises the following operations:
s21, carrying out ultrasonic treatment on the cut three-dimensional bacterial cellulose membrane in an absolute ethyl alcohol solution for 15-20min, and then carrying out repeated ultrasonic treatment in ultrapure water for 15-20min each time;
s22, soaking in ultrapure water for standby, and forming a hydrogel-textured film;
and S23, taking the three-dimensional bacterial cellulose membrane processed in the step S22, finally soaking the three-dimensional bacterial cellulose membrane in an ammonia water solution, carrying out ultrasonic treatment for 30-35min, and standing for later use.
Further, the ammonia water concentration of the soaking film in the step S23 is 5-100mM.
Further, the substrate preparation in step S3 specifically includes the following operations: measuring a silver salt solution, heating in a container, directly adding the soaked three-dimensional bacterial cellulose membrane into the silver salt solution, continuously heating for silver mirror reaction until the color of the three-dimensional bacterial cellulose membrane is changed from white to brown black, stopping the reaction, taking out the membrane, repeatedly washing the membrane with distilled water, and finally drying in a vacuum drying oven to prepare the Ag NPs-BCM substrate.
Further, the silver salt solution is CF 3 COOAg solution with the concentration of 5-100mM; the silver mirror reaction is carried out under the condition of keeping out of the sun, the heating temperature is 30-80 ℃, and the reaction color change time of the bacterial cellulose membrane is 1-9h.
It is a second object of the present invention to provide an Ag NPs-BCM substrate prepared according to the above method.
The invention also aims to provide an application of the Ag NPs-BCM substrate in cancer marker SERS detection, DTNB is added into serum to be specifically combined with a cancer marker of the Ag NPs-BCM substrate, the Ag NPs-BCM substrate according to claim 9 is fully contacted with cancer marker glutathione, and the cancer marker glutathione is detected by using an SERS method.
Compared with the prior art, the invention has the beneficial effects that:
1. according to the invention, the unique three-dimensional network structure and performance of the bacterial cellulose membrane are utilized, and the nano silver is uniformly loaded on the bacterial cellulose membrane by an in-situ reduction method, so that the agglomeration of nano colloids is solved to a certain extent, and no additional reducing agent is required to be added in the process; the soaking ammonia water is beneficial to weak hydrolysis of cellulose and combination with nano-cellulose of BCM, gives play to the advantages of the cellulose as a stabilizer and a reducing agent, and provides a reliable combination site for reducing silver nano-particles in the next step; in addition, in the dehydration and drying process of the prepared bacterial cellulose membrane containing the silver nanoparticles in vacuum drying, under the condition that the quantity of the silver nanoparticles is unchanged, the volume of the wet bacterial cellulose membrane is gradually reduced along with the evaporation of water, the gaps among the silver nanoparticles are further reduced, and SERS hot spots are increased;
2. the invention is in silver saltsThe silver trifluoroacetate has certain catalytic effect on metal ions, the adsorption of silver nanoparticles on cellulose chains is further increased, and the use risk is lower than that of AgNO 3 ；
3. The invention firstly soaks ammonia solution to make ammonia molecule and hydroxyl on bacterial cellulose membrane closely connected through hydrogen bond, and then CF 3 In COOAg, silver ions and ammonia water form a silver-ammonia solution, cellulose is used for in-situ reduction, the operation is safe and simple, and the prepared substrate is easy to store and carry; only need use very little volume SERS basement to soak the sample that awaits measuring when detecting glutathione, take out the drying after a period, can directly carry out SERS and measure, need not carry out extra preliminary treatment, labour saving and time saving to the sample that awaits measuring.
Reference numerals
FIG. 1 is a schematic diagram of the preparation method of Ag NPs-BCM substrate and its application in cancer marker detection according to the present invention;
FIG. 2 shows a 25 π cm sample prepared according to the present invention 2 Optical pictures of bacterial cellulose membranes of (1);
FIG. 3 shows CF at different concentrations in example 1 of the present invention 3 Raman mapping of SERS basement membrane prepared from COOAg to 4-MBN;
FIG. 4 is a Raman plot of SERS substrate membrane versus 4-MBN prepared at different temperatures in example 2 of the present invention;
FIG. 5 is a Raman plot of SERS substrate membrane vs. 4-MBN prepared at different reaction times in example 3 of the present invention;
FIG. 6 is a field emission scanning electron microscope image of the Ag NPs-BCM substrate prepared in example 4 of the present invention;
FIG. 7 is a Raman chart of the SERS basement membrane prepared in example 4 of the invention for detecting glutathione in serum of 10 different gastric cancer patients;
FIG. 8 is a Raman chart of the SERS basement membrane prepared in example 4 of the invention for detecting glutathione in serum of 10 different colon cancer patients.
Detailed Description
The invention will be further described with reference to the accompanying drawings and preferred embodiments, which are given for illustration only and are not intended to limit the scope of the invention.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified;
the quantitative tests in the following examples, all set up three replicates and the results averaged.
The experimental methods in the following examples are all conventional methods unless otherwise specified;
EXAMPLE 1 CF at various concentrations 3 Preparation of Ag NPs-BCM substrate from COOAg and SERS substrate film
As shown in fig. 1, a method for preparing an Ag NPs-BCM substrate includes the steps of:
s1, preparation of a three-dimensional bacterial cellulose membrane: in this example, a three-dimensional bacterial cellulose membrane was prepared using a green tea fermentation broth; firstly, 6g of green tea fermentation liquor is added into 700mL of boiling water, and the green tea fermentation liquor is fully stirred; secondly, standing the fermentation liquor, separating to obtain filtrate, adding 40g of white granulated sugar into the filtrate, and fully stirring to obtain sugar green tea fermentation liquor; finally, after the sugar green tea fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 300mL of green tea fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator at 30 ℃ to obtain a three-dimensional bacterial cellulose membrane;
s2, pretreatment of the three-dimensional bacterial cellulose membrane: the three-dimensional bacterial cellulose membrane prepared in the step S1 is cut, and the three-dimensional bacterial cellulose membrane prepared in the embodiment is 25 pi cm 2 Cutting to obtain 5 bacterial cellulose membranes, firstly carrying out ultrasonic treatment on the cut bacterial cellulose membranes in an absolute ethyl alcohol solution for 15min, and then repeatedly carrying out ultrasonic treatment in ultrapure water for 2-3 times, wherein the ultrasonic treatment is carried out for 15min each time; after the ultrasonic treatment, soaking the membrane in ultrapure water for later use to form a hydrogel-textured membrane; soaking the soaked bacterial cellulose membrane in 50mM ammonia water solution, performing ultrasonic treatment for 30min, and standing for 2h for later use to finish pretreatment of the three-dimensional bacterial cellulose membrane;
s3, substrate preparation: adding the three-dimensional bacterial cellulose membrane prepared in the step S2 into silver salt solution, and passing through a silver mirrorReducing silver salt solution in a three-dimensional bacterial cellulose membrane to prepare an Ag NPs-BCM substrate rich in silver nanospheres; in this example, CF of different concentrations was used 3 Preparing a substrate by COOAg, which comprises the following specific steps:
s31, 15mL in measured volume, 5mM,25mM,50mM,75mM, and 100mM CF, respectively 3 Respectively filling COOAg solutions into conical flasks, completely wrapping with tinfoil for light-shielding treatment, and heating in a water bath kettle at 65 deg.C;
s32, soaking the three-dimensional bacterial cellulose membrane in ammonia water, and directly adding the soaked three-dimensional bacterial cellulose membrane into CF 3 Adjusting the rotation speed of a water bath kettle in a COOAg solution, reacting for 4 hours in a dark condition, stopping the reaction, taking out the membrane, washing the membrane for multiple times by using ultrapure water, placing the membrane on a glass sheet, drying the glass sheet for 2 hours in a vacuum drying oven, carefully peeling off an SERS (surface enhanced Raman scattering) basement membrane which is numbered as 5mM,25mM,50mM,75mM and 100mM in sequence;
s33, respectively taking the 5 SERS basement membranes prepared above, wherein the areas of the 5 SERS basement membranes are 5x5mm 2 Directly soaking in 1000nM 4-MBN (4-mercaptobenzonitrile) overnight, taking out the membrane, drying in an oven at 30 ℃ for 30min, and directly carrying out SERS measurement.
As can be seen from FIG. 3, the concentration of CF was 50mM in the range of 5mM-100mM 3 The SERS basement membrane prepared from COOAg solution has the strongest signal for 4-MBN.
EXAMPLE 2 preparation of Ag NPs-BCM substrates as SERS substrate films at different temperatures
A preparation method of an Ag NPs-BCM substrate comprises the following steps:
s1, preparation of a three-dimensional bacterial cellulose membrane: in this example, a three-dimensional bacterial cellulose membrane was prepared using a green tea fermentation broth; firstly, adding 3g of green tea fermentation liquor into 500mL of boiling water, and fully stirring the green tea fermentation liquor; secondly, standing the fermentation liquor, separating to obtain filtrate, adding 20g of white granulated sugar into the filtrate, and fully stirring to obtain sugar green tea fermentation liquor; finally, after the sugar green tea fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 100mL of green tea fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator at 32 ℃ to obtain a three-dimensional bacterial cellulose membrane;
s2, three-dimensional bacterial cellulose membrane pretreatment: cutting the three-dimensional bacterial cellulose membrane prepared in the step S1, carrying out ultrasonic treatment on the cut bacterial cellulose membrane in an absolute ethyl alcohol solution for 20min, and then repeatedly carrying out ultrasonic treatment in ultrapure water for 2-3 times, wherein the ultrasonic treatment is carried out for 20min each time; after the ultrasonic treatment, soaking the membrane in ultrapure water for later use to form a hydrogel-textured membrane; soaking the soaked bacterial cellulose membrane in 5mM ammonia water solution, performing ultrasonic treatment for 33min, and standing for 2h for later use to finish pretreatment of the three-dimensional bacterial cellulose membrane;
s3, substrate preparation: adding the three-dimensional bacterial cellulose membrane prepared in the step S2 into a silver salt solution, and reducing the silver salt solution on the three-dimensional bacterial cellulose membrane through silver mirror reaction to prepare an Ag NPs-BCM substrate rich in silver nanospheres; in this example, the preparation of the Ag NPs-BCM substrate was carried out at different temperatures, and the specific steps were as follows:
s31, the measured volume is 15mL, and the concentration is 50mM CF 3 Respectively filling COOAg solutions into conical flasks, completely wrapping with tinfoil for light-shielding treatment, and heating in a water bath kettle at 30 deg.C, 50 deg.C, 60 deg.C, 65 deg.C, 70 deg.C, and 80 deg.C;
s32, soaking a bacterial cellulose membrane in ammonia water, and directly adding the bacterial cellulose membrane into CF 3 Adjusting the rotation speed of a water bath kettle in a COOAg solution, reacting for 4 hours in a dark condition, stopping the reaction, taking out a film, washing the film for multiple times by using ultrapure water, placing the film on a glass sheet, drying the film for 2 hours in a vacuum drying oven, and carefully peeling off an SERS (surface enhanced Raman scattering) substrate film, wherein the numbers are 30 ℃,50 ℃,60 ℃,65 ℃,70 ℃ and 80 ℃ in sequence;
s33, respectively taking the 5 SERS basement membranes prepared above, wherein the areas of the 5 SERS basement membranes are 5x5mm 2 Directly soaking in 1000nM 4-MBN (4-mercaptobenzonitrile) overnight, taking out the membrane, drying in an oven at 30 ℃ for 30min, and directly carrying out SERS measurement.
As can be seen from FIG. 4, the SERS basement membrane prepared by stabilizing the temperature at 65 ℃ has the strongest signal for 4-MBN in the range of 30-80 ℃.
Example 3 preparation of Ag NPs-BCM substrates as SERS substrate films at different reaction times
A preparation method of an Ag NPs-BCM substrate comprises the following steps:
s1, preparation of a three-dimensional bacterial cellulose membrane: in this example, a three-dimensional bacterial cellulose membrane was prepared using a green tea fermentation broth; firstly, adding 10g of green tea fermentation liquor into 1000mL of boiling water, and fully stirring the green tea fermentation liquor; secondly, standing the fermentation liquor, separating to obtain filtrate, adding 60g of white granulated sugar into the filtrate, and fully stirring to obtain sugar green tea fermentation liquor; finally, after the sugar green tea fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 500mL of green tea fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator at 35 ℃ to obtain a three-dimensional bacterial cellulose membrane;
s2, three-dimensional bacterial cellulose membrane pretreatment: cutting the three-dimensional bacterial cellulose membrane prepared in the step S1, carrying out ultrasonic treatment on the cut bacterial cellulose membrane in an absolute ethyl alcohol solution for 18min, and then repeatedly carrying out ultrasonic treatment in ultrapure water for 2-3 times, wherein the ultrasonic treatment is carried out for 18min each time; after the ultrasonic treatment, soaking the membrane in ultrapure water for later use to form a hydrogel-textured membrane; soaking the soaked bacterial cellulose membrane in 100mM ammonia water solution, performing ultrasonic treatment for 35min, and standing for 2h for later use to finish pretreatment of the three-dimensional bacterial cellulose membrane;
s3, preparing a substrate: adding the three-dimensional bacterial cellulose membrane prepared in the step S2 into a silver salt solution, and reducing the silver salt solution in the three-dimensional bacterial cellulose membrane through silver mirror reaction to prepare an Ag NPs-BCM substrate rich in silver nanospheres; in the embodiment, the Ag NPs-BCM substrate is prepared in different reaction times, and the specific steps are as follows;
s31, the measured volume is 15mL, and the concentration is 50mM CF 3 Respectively filling COOAg solutions into conical flasks, completely wrapping with tinfoil for light-shielding treatment, and heating in a water bath kettle at 65 deg.C;
s32, soaking the bacterial cellulose membrane in ammonia water, and directly adding the bacterial cellulose membrane into CF 3 Adjusting the rotation speed of a water bath kettle in a COOAg solution, sequentially reacting for 1h,3h,4h,5h,7h and 9h under the condition of keeping out of the light, stopping the reaction, taking out the membrane, washing the membrane with ultrapure water for multiple times, placing the membrane on a glass sheet, drying the membrane for 2h in a vacuum drying box, carefully peeling off an SERS (surface enhanced Raman Scattering) basement membrane which is numbered as 1h,3h,4h,5h,7h and 9h in sequence;
s33, respectively taking the 5 SERS basement membranes prepared above, wherein the areas of the 5 SERS basement membranes are 5x5mm 2 Directly soaking in 1000nM 4-MBN (4-mercaptobenzonitrile) overnight, taking out the membrane, drying in a 30 ℃ oven for 30min, and directly carrying out SERS measurement;
as can be seen from FIG. 5, in the range of 1h to 9h, the SERS basement membrane prepared at the reaction time of 4h has the strongest signal for 4-MBN; FIG. 6 shows the optimal conditions (at 50mM CF) 3 COOAg, reaction at 65 ℃ for 4 h) field emission Scanning Electron Microscopy (SEM) of the prepared Ag NPs-BCM substrate.
Example 4 detection of glutathione by SERS basement membrane in serum of gastric cancer patients
According to the exploration of experimental conditions in the process of preparing the SERS substrate in the embodiment 1-3, the optimal experimental condition combination for the 4-MBN signal is found; under the combined condition, detecting glutathione in the serum of a gastric cancer patient by using an SERS basement membrane; among them, the optimum experimental conditions are as follows:
(1) Preparation of three-dimensional bacterial cellulose membrane:
s11, adding 6g of green tea fermentation liquor into 700mL of boiling water, and fully stirring the green tea fermentation liquor;
s12, standing the fermentation liquor, separating to obtain a filtrate, adding 40g of white granulated sugar into the filtrate, and fully stirring to obtain sugar green tea fermentation liquor;
s13, after the sugar green tea fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 300mL of green tea fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator at 30 ℃ to obtain three-dimensional bacterial cellulose;
(2) Pretreating a three-dimensional bacterial cellulose membrane:
s21, cutting area of 25 pi cm 2 The bacterial cellulose membrane is subjected to ultrasonic treatment in an absolute ethyl alcohol solution for 15min, and then ultrasonic treatment is repeatedly carried out in ultrapure water for 2-3 times, and each ultrasonic treatment is carried out for 15min;
s22, soaking the membrane in ultrapure water for later use to form a hydrogel-textured membrane;
s23, soaking the treated bacterial cellulose membrane in 50mM ammonia water solution, firstly carrying out ultrasonic treatment for 30min, and then standing for 2h for later use;
(3) Preparing a substrate:
s31, the measured volume is 15mL, and the concentration is 50mM CF 3 Respectively filling COOAg solutions into conical flasks, completely wrapping with tinfoil for light-shielding treatment, and heating in a water bath kettle at 65 deg.C;
s32, soaking the bacterial cellulose membrane in ammonia water, and directly adding the bacterial cellulose membrane into CF 3 Adjusting the rotation speed of a water bath kettle in a COOAg solution, reacting for 4 hours under the condition of keeping out of the sun, stopping the reaction, taking out the membrane, washing the membrane for multiple times by using ultrapure water, placing the membrane on a glass sheet, drying for 2 hours in a vacuum drying oven, and carefully peeling off an SERS (surface enhanced Raman Scattering) base membrane;
the SERS basement membrane prepared by the method is used for detecting cancer markers, 10 mu L of serum of samples of normal people, gastric cancer and colon cancer are sequentially sucked into a centrifuge tube, 10 samples (from different people) are respectively taken from each sample, 80 mu L of PBS buffer solution with the same volume is sequentially added, and the mixture is shaken for 3min on a shaking table at the temperature of 30 ℃. Finally, sequentially adding 10 mu L of 10mM DTNB, and continuously incubating for 30min on a shaking table at 30 ℃ to prepare a sample to be detected; sucking 100 mu L of sample to be detected in a centrifuge tube, and taking the prepared SERS basement membrane with the thickness of 5x5mm 2 And a plurality of the films are directly soaked in a centrifuge tube overnight, the films are taken out and dried in an oven at 30 ℃ for 30min, and SERS measurement is directly carried out.
As can be seen from fig. 7 and 8, the mean GSH levels were higher in colon cancer patients and stomach cancer patients than in normal persons by directly adding DTNB to the serum; by calculation, the average GSH level of colon cancer patients is 2.6 times of that of normal people, and the average GSH level of stomach cancer patients is 2.8 times of that of normal people; therefore, the GSH content of cancer patients and normal people can be directly detected by adding DTNB into serum and using Ag NPs-BCM; the method provided by the application can simply and rapidly distinguish cancer patients from normal people, and takes GSH as a potential cancer marker.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (10)
1. A preparation method of an Ag NPs-BCM substrate is characterized by comprising the following steps: after the bacterial cellulose membrane is soaked in ammonia water for volume contraction treatment, ag NPs with high density are grown in situ on the three-dimensional bacterial cellulose membrane by utilizing silver mirror reaction and are uniformly distributed, and then the Ag NPs-BCM substrate rich in silver nanospheres is prepared.
2. The method of preparing an Ag NPs-BCM substrate according to claim 1, comprising the steps of:
s1, preparation of a three-dimensional bacterial cellulose membrane: preparing a three-dimensional bacterial cellulose membrane by using fermentation liquor;
s2, three-dimensional bacterial cellulose membrane pretreatment: sequentially soaking the three-dimensional bacterial cellulose membrane prepared in the step S1 in ultrapure water, absolute ethyl alcohol and ammonia water for pretreatment to form a three-dimensional bacterial cellulose membrane with a hydrogel texture;
s3, substrate preparation: and (3) adding the three-dimensional bacterial cellulose membrane prepared in the step (S2) into the silver salt solution, and reducing the silver salt solution in the three-dimensional bacterial cellulose membrane through silver mirror reaction to prepare the Ag NPs-BCM substrate rich in silver nanospheres.
3. The method for preparing the Ag NPs-BCM substrate according to claim 2, wherein the preparation of the three-dimensional bacterial cellulose membrane in the step S1 is specifically performed as follows:
s11, adding 3-10g of fermentation liquor into 500-1000mL of boiling water, and fully stirring the fermentation liquor;
s12, standing the fermentation liquor, separating to obtain a filtrate, adding 20-60g of white granulated sugar into the filtrate, and fully stirring to obtain a sugar fermentation liquor;
and S13, after the sugar fermentation liquor is cooled, supplementing the original volume with ultrapure water, adding 100-500mL of fermentation bacteria mother liquor, uniformly mixing and sealing, and fermenting in a constant-temperature incubator to obtain the three-dimensional bacterial cellulose membrane.
4. The method for preparing an Ag NPs-BCM substrate according to claim 3, wherein the fermentation broth is green tea fermentation broth; the fermentation bacteria mother liquor is green tea bacteria mother liquor; the temperature of the constant temperature incubator is 30-35 ℃.
5. The method for preparing the Ag NPs-BCM substrate according to claim 2, wherein the three-dimensional bacterial cellulose membrane pretreatment in the step S2 is carried out by the following steps:
s21, cutting the prepared three-dimensional bacterial cellulose membrane, performing ultrasonic treatment in an absolute ethyl alcohol solution for 15-20min, and then performing repeated ultrasonic treatment in ultrapure water for 15-20min each time;
s22, soaking in ultrapure water for standby, and forming a hydrogel-textured film;
and S23, taking the three-dimensional bacterial cellulose membrane processed in the step S22, finally soaking the three-dimensional bacterial cellulose membrane in an ammonia water solution, carrying out ultrasonic treatment for 30-35min, and standing for later use.
6. The method for preparing an Ag NPs-BCM substrate according to claim 5, wherein the ammonia concentration of the soaking film in the step S23 is 5-100mM.
7. The method for preparing an Ag NPs-BCM substrate according to claim 2, wherein the substrate preparation in step S3 is performed by: measuring a silver salt solution, heating in a container, directly adding the soaked three-dimensional bacterial cellulose membrane into the silver salt solution, continuously heating for silver mirror reaction until the color of the three-dimensional bacterial cellulose membrane is changed from white to brown black, stopping the reaction, taking out the membrane, repeatedly washing the membrane with distilled water, and finally drying in a vacuum drying oven to prepare the Ag NPs-BCM substrate.
8. The method of claim 7, wherein the silver salt solution is CF 3 COOAg solution with the concentration of 5-100mM; the silver mirror reaction is carried out under the condition of keeping out of the sun, the heating temperature is 30-80 ℃, and the reaction color change time of the bacterial cellulose membrane is 1-9h.
9. An Ag NPs-BCM substrate produced according to the process of any one of claims 1 to 8.
10. The application of the Ag NPs-BCM substrate in cancer marker SERS detection is characterized in that DTNB is added into serum to be specifically combined with a cancer marker, the Ag NPs-BCM substrate as claimed in claim 9 is fully contacted with cancer marker glutathione, and the cancer marker glutathione is detected by using an SERS method.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090220560A1 (en) * | 2006-04-24 | 2009-09-03 | Axcelon Biopolymers Corporation | Nanosilver Coated Bacterial Cellulose |
| CN102206355A (en) * | 2011-03-28 | 2011-10-05 | 东华大学 | Silver nanoparticle-containing bacterial cellulose membrane and preparation method thereof |
| CN109884031A (en) * | 2019-03-14 | 2019-06-14 | 中国人民解放军军事科学院军事医学研究院 | The method for detecting reduced glutathione and/or oxidized form of glutathione |
| CN109944067A (en) * | 2019-03-05 | 2019-06-28 | 华南理工大学 | A kind of nano silver particles/cellulose fiber composite material and preparation method |
| CN113155805A (en) * | 2021-03-11 | 2021-07-23 | 福建师范大学 | Cellulose-based SERS substrate based on Tollens reaction and preparation method thereof |
| CN114994311A (en) * | 2022-07-13 | 2022-09-02 | 福建师范大学 | SERS technology-based immune lateral flow test strip with self-calibration function and preparation method thereof |
-
2022
- 2022-09-14 CN CN202211116151.4A patent/CN115612156B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090220560A1 (en) * | 2006-04-24 | 2009-09-03 | Axcelon Biopolymers Corporation | Nanosilver Coated Bacterial Cellulose |
| CN102206355A (en) * | 2011-03-28 | 2011-10-05 | 东华大学 | Silver nanoparticle-containing bacterial cellulose membrane and preparation method thereof |
| CN109944067A (en) * | 2019-03-05 | 2019-06-28 | 华南理工大学 | A kind of nano silver particles/cellulose fiber composite material and preparation method |
| CN109884031A (en) * | 2019-03-14 | 2019-06-14 | 中国人民解放军军事科学院军事医学研究院 | The method for detecting reduced glutathione and/or oxidized form of glutathione |
| CN113155805A (en) * | 2021-03-11 | 2021-07-23 | 福建师范大学 | Cellulose-based SERS substrate based on Tollens reaction and preparation method thereof |
| CN114994311A (en) * | 2022-07-13 | 2022-09-02 | 福建师范大学 | SERS technology-based immune lateral flow test strip with self-calibration function and preparation method thereof |
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