Formula of melanoma decolorizing agent
By combining the use of hydrogen peroxide, borate buffer, Tris-HCl buffer, EDTA and carboxymethyl chitosan solution, the problems of long decolorization time and unstable effect in existing melanoma decolorization methods are solved, achieving rapid and stable melanoma decolorization and reducing damage to tissues.
Patent Information
- Application Number
- CN202510803001.8
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-16
- Publication Date
- 2025-09-23
AI Technical Summary
Existing melanoma decolorization methods have problems such as long decolorization time, unstable decolorization effect, acid-base environment affecting immunohistochemical staining, and high temperature causing tissue structure damage.
A combination of hydrogen peroxide, borate buffer, Tris-HCl buffer, EDTA and carboxymethyl chitosan solution was used to enhance the oxidative activity and stability by adjusting the pH value and chelating metal ions, forming a physical barrier to inhibit the decomposition of hydrogen peroxide and improve the decolorization efficiency.
Rapid and stable melanoma decolorization is achieved, damage to tissues is reduced, and the stability and decolorization effect of the decolorizer are enhanced.
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of decolorizing agents, and in particular to a melanoma decolorizing agent formula. Background Art
[0002] Melanoma, also known as melanoma or malignant melanoma, is a highly malignant tumor that originates from melanocytes in the skin or mucous membranes. It is highly invasive and prone to metastasis. The disease is primarily caused by excessive ultraviolet light exposure, genetic factors, and immune system abnormalities. Patients typically present with irregularly shaped, varying shades of plaque or nodule on the skin or mucous membranes, which may be accompanied by itching, pain, or bleeding. If left untreated, melanoma can rapidly spread throughout the body, leading to serious complications and even life-threatening conditions.
[0003] The earliest melanin depigmentation technology began in 1897, initially using 0.1% potassium permanganate for oxidative depigmentation. Subsequently, various depigmentation methods were developed under varying experimental conditions. However, only two methods are commonly used due to their superior depigmentation efficacy and ease of use. These two methods, potassium permanganate-oxalic acid and hydrogen peroxide, each have their own limitations, including prolonged depigmentation times, unstable depigmentation, acid-base conditions that affect subsequent immunohistochemical staining, and high temperatures that can damage tissue structure and cause staining to flake. Therefore, we propose a melanoma depigmentation agent formulation. Summary of the Invention
[0004] The purpose of the present invention is to solve the problems mentioned in the above background technology, and the present invention provides a melanoma depigmentation agent formula.
[0005] In order to achieve the above-mentioned purpose, the present invention specifically adopts the following technical solutions:
[0006] A melanoma depigmenting agent formulation is prepared from the following components by mass: 7 mL of hydrogen peroxide, 0.7 mL of borate buffer, 0.7 mL of Tris-HCl buffer, 0.2 mL of EDTA, and 0.1 mL of carboxymethyl chitosan solution.
[0007] Furthermore, the hydrogen peroxide is used to provide oxidative active ingredients; the borate buffer is used to maintain a pH value of 10; the Tris-HCl buffer is used to accurately adjust the pH to a target value; the EDTA is used to inhibit the catalytic decomposition of hydrogen peroxide; and the carboxymethyl chitosan solution is used to enhance metal chelation, scavenge free radicals, and form a physical barrier.
[0008] Furthermore, the initial concentration of the hydrogen peroxide solution is 3%.
[0009] Furthermore, the method for preparing the borate buffer comprises the following steps:
[0010] Step 11: Prepare a 0.1 mol / L boric acid solution by weighing 1.24 g of boric acid (molar mass 39.91 g / mol) and dissolving it in 100 mL of deionized water. Stir thoroughly until completely dissolved.
[0011] Step 12: Prepare a 0.1 mol / L borax solution: weigh 1.91 g of borax (molar mass 381.43 g / mol), dissolve it in 100 mL of deionized water, and stir thoroughly until completely dissolved.
[0012] Step 13, mixing and preparing a buffer solution: according to the required pH value, mix the above two solutions in proportion;
[0013] Step 14. Volume adjustment and storage: Transfer the mixed buffer solution to a 100 mL volumetric flask, dilute to the mark with deionized water, and store in a sealed container at room temperature or in a refrigerated environment to prevent deterioration.
[0014] Furthermore, the preparation method of the Tris-HCl buffer comprises the following steps:
[0015] Step 21, raw material preparation: select tris(hydroxymethyl)aminomethane, concentrated hydrochloric acid and distilled water as raw materials;
[0016] Step 22, preliminary mixing: Mix the Tris solution of the required concentration with hydrochloric acid in a volume ratio and stir thoroughly to dissolve;
[0017] Step 23, pH adjustment: Use a pH meter to measure the pH value of the mixed solution and adjust the amount of hydrochloric acid added according to the target pH.
[0018] Furthermore, the configuration method of the EDTA comprises the following steps:
[0019] Step 31, reagent preparation: analytical grade EDTA (such as disodium EDTA) or EDTA standard solution, deionized water or distilled water;
[0020] Step 32, dissolving: Weigh EDTA solid into a beaker, add about 200 mL of deionized water, and dissolve under slight heat;
[0021] Step 33. Storage: After cooling to room temperature, transfer to a 1000 mL volumetric flask, add distilled water to the mark, and shake well for later use.
[0022] Furthermore, the degree of substitution of the carboxymethyl chitosan is 80%.
[0023] Furthermore, the preparation method of the carboxymethyl chitosan solution comprises the following steps:
[0024] Step 41, materials and reagents: chitosan powder is used as raw material, and isopropyl alcohol, sodium hydroxide solution and chloroacetic acid are used as reagents;
[0025] Step 42, swelling: taking chitosan powder and adding it into isopropyl alcohol, stirring and soaking it to allow it to fully swell;
[0026] Step 43, alkalization: add sodium hydroxide solution, adjust the pH to 10-12, and react in a water bath at 20-60°C for 2-8 hours to ensure complete alkalization;
[0027] Step 44, carboxymethylation: dissolve chloroacetic acid in isopropyl alcohol, add the mixture to the reaction system in portions, and react in a water bath at 50-70° C. for 3-4 hours, stirring during the reaction;
[0028] Step 45, reaction termination and purification: After the reaction is completed, adjust the pH to 10 with glacial acetic acid to terminate the reaction, add distilled water in portions to fully dissolve the residue, centrifuge and separate the crude product, and wash the precipitate with 75% to 80% ethanol or methanol several times to remove unreacted chloroacetic acid and salts.
[0029] Furthermore, the concentration of isopropyl alcohol in the materials and reagents is 70% to 90%, the concentration of sodium oxide solution is 10% to 30%, and the concentration of chloroacetic acid is 30% to 50%.
[0030] Further, the following steps are included:
[0031] Step A, mixing: mix hydrogen peroxide, borate, Tris-HCl and EDTA solution in proportion and stir gently;
[0032] Step B, stirring: slowly add CMCS solution to avoid introducing bubbles and stir until uniform;
[0033] Step C, detection: Detect the final pH and adjust with trace amounts of HCl / NaOH if necessary;
[0034] Step D: Aliquot into brown glass bottles and store at 4°C away from light.
[0035] The beneficial effects of the present invention are as follows:
[0036] 1. The initial concentration of hydrogen peroxide in the present invention is 3%. 3% hydrogen peroxide is the core active ingredient and its concentration needs to be maintained. Hydrogen peroxide is used to provide the oxidative active ingredient. Borate buffer is used to maintain the alkaline environment of the system with a pH value of 10. Tris-HCl buffer is used to accurately adjust the pH to the target value and enhance the buffering capacity. EDTA is used to inhibit the catalytic decomposition of hydrogen peroxide and eliminate impurities caused by the catalytic decomposition. Carboxymethyl chitosan solution is used to enhance metal chelation, scavenge free radicals and form a physical barrier.
[0037] 2. The carboxymethyl chitosan of the present invention contains carboxyl and amino groups, which can chelate trace metal ions and reduce their catalytic effect on the decomposition of hydrogen peroxide. The formation of gel or film by carboxymethyl chitosan may hinder the release of oxygen and slow down the decomposition kinetics. On the basis of EDTA, carboxymethyl chitosan can further chelate residual metal ions and enhance the stabilization effect. EDTA can react with Fe 2+ 、Cu 2+ EDTA combines with metal ions to reduce its catalytic effect on the decomposition of hydrogen peroxide (H2O2). The chelation effect of EDTA is best at pH 8-10, which matches the pH 10 of Tris-HCl buffer. After EDTA chelates metal ions, the decomposition rate of hydrogen peroxide is significantly slowed down.
[0038] 3. The carboxymethyl chitosan with a degree of substitution of 80 in the present invention generates more hydroxyl radicals, thereby enhancing the oxidative decolorization ability; the pH of the carboxymethyl chitosan solution with a degree of substitution of 80 is closer to neutral or weakly alkaline, which promotes the decomposition of hydrogen peroxide, generates more active free radicals, and improves the decolorization efficiency; the carboxymethyl chitosan solution with a degree of substitution of 80 has a low viscosity, which is conducive to the contact between free radicals and pigments. DETAILED DESCRIPTION
[0039] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below.
[0040] The invention provides a melanoma decolorizing agent formula, which is prepared from the following components by weight: 7 mL of hydrogen peroxide, 0.7 mL of borate buffer, 0.7 mL of Tris-HCl buffer, 0.2 mL of EDTA, and 0.1 mL of carboxymethyl chitosan solution.
[0041] The initial concentration of hydrogen peroxide is 3%. 3% hydrogen peroxide is the core active ingredient and its concentration needs to be maintained. Hydrogen peroxide is used to provide oxidative active ingredients. Borate buffer is used to maintain the alkaline environment of the system with a pH of 10. Tris-HCl buffer is used to accurately adjust the pH to the target value and enhance buffering capacity. EDTA is used to inhibit the catalytic decomposition of hydrogen peroxide and eliminate impurities caused by catalytic decomposition. Carboxymethyl chitosan solution is used to enhance metal chelation, scavenge free radicals and form a physical barrier.
[0042] Carboxymethyl chitosan contains carboxyl and amino groups, which can chelate trace metal ions (such as Fe 2+ 、Cu 2+), reducing its catalytic effect on the decomposition of hydrogen peroxide, good metal chelating ability, chitosan derivatives have a certain free radical scavenging ability, which may inhibit the chain reaction of active oxygen (such as OH) produced during the decomposition of hydrogen peroxide, and have strong antioxidant properties. Carboxymethyl chitosan forms a gel or film, which may hinder the release of oxygen and slow down the decomposition kinetics. On the basis of EDTA, carboxymethyl chitosan can further chelate residual metal ions and enhance the stabilization effect; EDTA can further chelate residual metal ions and enhance the stabilization effect by reacting with Fe 2+ 、Cu 2+ EDTA combines with metal ions to reduce its catalytic effect on the decomposition of hydrogen peroxide (H2O2). The chelation effect of EDTA is best at pH 8-10, which matches the pH 10 of Tris-HCl buffer. After EDTA chelates metal ions, the decomposition rate of hydrogen peroxide is significantly slowed down.
[0043] In this embodiment, preferably, the preparation method of the borate buffer comprises the following steps:
[0044] Step 11: Prepare a 0.1 mol / L boric acid solution by weighing 1.24 g of boric acid (molar mass 39.91 g / mol) and dissolving it in 100 mL of deionized water. Stir thoroughly until completely dissolved.
[0045] Step 12: Prepare a 0.1 mol / L borax solution: weigh 1.91 g of borax (molar mass 381.43 g / mol), dissolve it in 100 mL of deionized water, and stir thoroughly until completely dissolved.
[0046] Step 13, mixing and preparing a buffer solution: according to the required pH value, mix the above two solutions in proportion;
[0047] Step 14. Volume adjustment and storage: Transfer the mixed buffer solution to a 100 mL volumetric flask, dilute to the mark with deionized water, and store in a sealed container at room temperature or in a refrigerated environment to prevent deterioration.
[0048] In this embodiment, preferably, the preparation method of Tris-HCl buffer comprises the following steps:
[0049] Step 21, raw material preparation: select tris(hydroxymethyl)aminomethane, concentrated hydrochloric acid and distilled water as raw materials;
[0050] Step 22, preliminary mixing: Mix the Tris solution of the required concentration with hydrochloric acid in a volume ratio and stir thoroughly to dissolve;
[0051] Step 23, pH adjustment: Use a pH meter to measure the pH value of the mixed solution and adjust the amount of hydrochloric acid added according to the target pH.
[0052] In this embodiment, preferably, the preparation method of EDTA comprises the following steps:
[0053] Step 31, reagent preparation: analytical grade EDTA (such as disodium EDTA) or EDTA standard solution, deionized water or distilled water;
[0054] Step 32, dissolving: Weigh EDTA solid into a beaker, add about 200 mL of deionized water, and dissolve under slight heat;
[0055] Step 33. Storage: After cooling to room temperature, transfer to a 1000 mL volumetric flask, add distilled water to the mark, and shake well for later use.
[0056] In this embodiment, preferably, the degree of substitution of carboxymethyl chitosan is 80%.
[0057] Carboxymethylation mainly replaces the amino groups in the chitosan molecule. Carboxymethyl chitosan with a substitution degree of 80 retains more amino groups, which may catalyze the decomposition of hydrogen peroxide and generate more hydroxyl radicals, thereby enhancing the oxidative decolorization ability. Carboxymethyl is a weakly acidic group. Carboxymethyl chitosan with a high substitution degree will release more H ions, making the solution pH lower. The pH of the carboxymethyl chitosan solution with a substitution degree of 80 is closer to neutral or weakly alkaline, which promotes the decomposition of hydrogen peroxide, generates more active free radicals, and improves the decolorization efficiency. Carboxymethyl chitosan with a substitution degree of 80 retains some amino groups (positively charged), which can bind to pigments through electrostatic adsorption and assist in oxidative decolorization. The viscosity of the carboxymethyl chitosan solution with a substitution degree of 80 is low, which is conducive to the contact between free radicals and pigments.
[0058] In this embodiment, preferably, the method for preparing the carboxymethyl chitosan solution comprises the following steps:
[0059] Step 41, materials and reagents: chitosan powder is used as raw material, and isopropyl alcohol, sodium hydroxide solution and chloroacetic acid are used as reagents;
[0060] Step 42, swelling: taking chitosan powder and adding it into isopropyl alcohol, stirring and soaking it to allow it to fully swell;
[0061] Step 43, alkalization: add sodium hydroxide solution, adjust the pH to 10-12, and react in a water bath at 20-60°C for 2-8 hours to ensure complete alkalization;
[0062] Step 44, carboxymethylation: dissolve chloroacetic acid in isopropyl alcohol, add the mixture to the reaction system in portions, and react in a water bath at 50-70° C. for 3-4 hours, stirring during the reaction;
[0063] Step 45, reaction termination and purification: After the reaction is completed, adjust the pH to 10 with glacial acetic acid to terminate the reaction, add distilled water in portions to fully dissolve the residue, centrifuge and separate the crude product, and wash the precipitate with 75% to 80% ethanol or methanol several times to remove unreacted chloroacetic acid and salts.
[0064] In this embodiment, preferably, the concentration of isopropyl alcohol in the materials and reagents is 70% to 90%, the concentration of the sodium oxide solution is 10% to 30%, and the concentration of chloroacetic acid is 30% to 50%.
[0065] In this embodiment, preferably, the following steps are included:
[0066] Step A, mixing: mix hydrogen peroxide, borate, Tris-HCl and EDTA solution in proportion and stir gently;
[0067] Step B, stirring: slowly add CMCS solution to avoid introducing bubbles and stir until uniform;
[0068] Step C, detection: Detect the final pH and adjust with trace amounts of HCl / NaOH if necessary;
[0069] Step D, subpackaging: Subpack into brown glass bottles, protect from light, and store at 4°C; using brown bottles for storage can reduce photocatalytic decomposition, and storing at 4°C can further inhibit decomposition.
[0070] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A melanoma decolorizing agent formulation, characterized in that: It was prepared from the following masses of components: 7 mL of hydrogen peroxide, 0.7 mL of borate buffer, 0.7 mL of Tris-HCl buffer, 0.2 mL of EDTA, and 0.1 mL of carboxymethyl chitosan solution.
2. A melanoma decolorizing agent formulation according to claim 1, characterized in that: The hydrogen peroxide is used to provide oxidative active ingredients; the borate buffer is used to maintain a pH value of 10; the Tris-HCl buffer is used to accurately adjust the pH to a target value; the EDTA is used to inhibit the catalytic decomposition of hydrogen peroxide; and the carboxymethyl chitosan solution is used to enhance metal chelation, scavenge free radicals, and form a physical barrier.
3. The melanoma decolorizing agent formulation according to claim 1, characterized in that: The initial concentration of the hydrogen peroxide solution is 3%.
4. A melanoma decolorizing agent formulation according to claim 1, characterized in that: The method for preparing the borate buffer comprises the following steps: Step 11: Prepare a 0.1 mol / L boric acid solution: weigh 1.24 g of boric acid (molar mass 39.91 g / mol), dissolve it in 100 mL of deionized water, and stir thoroughly until completely dissolved. Step 12: Prepare a 0.1 mol / L borax solution: weigh 1.91 g of borax (molar mass 381.43 g / mol), dissolve it in 100 mL of deionized water, and stir thoroughly until completely dissolved. Step 13, mixing and preparing a buffer solution: according to the required pH value, mix the above two solutions in proportion; Step 14. Volume adjustment and storage: Transfer the mixed buffer solution to a 100 mL volumetric flask, dilute to the mark with deionized water, and store in a sealed container at room temperature or in a refrigerated environment to prevent deterioration.
5. The melanoma decolorizing agent formulation according to claim 1, characterized in that: The Tris-HCl The preparation method of the buffer solution includes the following steps: Step 21, raw material preparation: select tris(hydroxymethyl)aminomethane, concentrated hydrochloric acid and distilled water as raw materials; Step 22, preliminary mixing: Mix the Tris solution of the required concentration with hydrochloric acid in a volume ratio and stir thoroughly to dissolve; Step 23, pH adjustment: Use a pH meter to measure the pH value of the mixed solution and adjust the amount of hydrochloric acid added according to the target pH.
6. A melanoma decolorizing agent formulation according to claim 1, characterized in that: The configuration method of the EDTA comprises the following steps: Step 31, reagent preparation: analytical grade EDTA (such as disodium EDTA) or EDTA standard solution, deionized water or distilled water; Step 32, dissolving: Weigh EDTA solid into a beaker, add about 200 mL of deionized water, and dissolve under slight heat; Step 33. Storage: After cooling to room temperature, transfer to a 1000 mL volumetric flask, add distilled water to the mark, and shake well for later use.
7. The melanoma decolorizing agent formulation according to claim 1, characterized in that: The selected substitution degree of the carboxymethyl chitosan is 80%.
8. The melanoma decolorizing agent formulation according to claim 1, characterized in that: The preparation method of the carboxymethyl chitosan solution comprises the following steps: Step 41, materials and reagents: chitosan powder is used as raw material, and isopropyl alcohol, sodium hydroxide solution and chloroacetic acid are used as reagents; Step 42, swelling: taking chitosan powder and adding it into isopropyl alcohol, stirring and soaking it to allow it to fully swell; Step 43, alkalization: add sodium hydroxide solution, adjust the pH to 10-12, and react in a water bath at 20-60°C for 2-8 hours to ensure complete alkalization; Step 44, carboxymethylation: dissolve chloroacetic acid in isopropyl alcohol, add the mixture to the reaction system in portions, and react in a water bath at 50-70° C. for 3-4 hours, stirring during the reaction; Step 45, reaction termination and purification: After the reaction is completed, adjust the pH to 10 with glacial acetic acid to terminate the reaction, add distilled water in portions to fully dissolve the residue, centrifuge and separate the crude product, and wash the precipitate with 75% to 80% ethanol or methanol several times to remove unreacted chloroacetic acid and salts.
9. The melanoma decolorizing agent formulation according to claim 8, characterized in that: The concentration of isopropyl alcohol in the materials and reagents is 70% to 90%, the concentration of the sodium oxide solution is 10% to 30%, and the concentration of chloroacetic acid is 30% to 50%.
10. The melanoma decolorizing agent formulation according to claim 1, characterized in that: The steps include: Step A, mixing: mix hydrogen peroxide, borate, Tris-HCl and EDTA solution in proportion and stir gently; Step B, stirring: slowly add CMCS solution to avoid introducing bubbles and stir until uniform; Step C, detection: Detect the final pH and adjust with trace amounts of HCl / NaOH if necessary; Step D: Aliquot into brown glass bottles and store at 4°C away from light.