CN115612560A - A nucleic acid pollution scavenger containing Chinese medicinal extract as main ingredient - Google Patents

Abstract

The invention discloses a nucleic acid pollution scavenger taking traditional Chinese medicine extract as a main component, which comprises the following components in percentage by mass: 10-15% of traditional Chinese medicine extract, 0.4-2.0% of oxidant, 0.2-3.0% of chlorine-containing reagent, 0.2-4% of acid-containing reagent, 0.1-2% of stabilizer, 1-3% of surfactant, 5-10% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 20-60% of cepharanthine, 10-40% of chlorogenic acid and 30-50% of tea saponin in percentage by mass. The nucleic acid pollution scavenger taking the traditional Chinese medicine extract as the main component adopts the formula, adopts the traditional Chinese medicine extract to synergistically degrade DNA, has no odor, mild property and no irritation, is simple and convenient to operate, convenient to store, long in storage period, environment-friendly and safe, and has a good nucleic acid removing effect.

Description

A nucleic acid pollution scavenger containing Chinese medicinal extract as main ingredient

Technical Field

The invention belongs to the technical field of disinfection and cleaning, and particularly relates to a nucleic acid pollution scavenger taking a traditional Chinese medicine extract as a main component.

Background

PCR, known as Polymerase chain reaction (Polymerase chain reaction), is a molecular biology technique for amplifying a large number of specific DNA fragments in vitro by using the principle of DNA double-strand replication, and can rapidly amplify a large number of copies by using a trace amount of DNA as a template. The PCR technology is widely applied to more fields of biology and related disciplines, and has remarkable effect in the fields of medical research, criminal evidence collection, molecular archaeology and the like.

At the same time, however, the unique principle of PCR technology teaches that one molecule will generate billions of molecules after PCR reaction, and this high sensitivity means that during the PCR operation, the extremely slight contamination of the environment and laboratory with nucleic acids may cause a large amount of amplification of the amplified product, resulting in false positive amplification results. Common nucleic acid contaminations in nucleic acid laboratories include: contamination of amplification products, contamination of extracted nucleic acids, contamination of positive controls, contamination of foreign nucleic acids, and the like. To deal with such contaminated nucleic acids, researchers have proposed various treatment methods such as nuclease degradation, ultraviolet irradiation, chloride oxidation, high temperature treatment, sodium hypochlorite soaking or scrubbing. However, some of these treatments have serious odors that affect human health and some have the risk of corroding and damaging the instrument.

Disclosure of Invention

The invention aims to provide a nucleic acid pollution scavenger taking traditional Chinese medicine extracts as main components, which aims to solve the problems of serious smell, strong corrosivity and the like existing in the nucleic acid pollution treatment method and further seek an effective, safe, rapid and stable nucleic acid pollution scavenger.

In order to achieve the aim, the invention provides a nucleic acid pollution scavenger taking a traditional Chinese medicine extract as a main component, which comprises the following components in percentage by mass: 10-15% of traditional Chinese medicine extract, 0.4-2.0% of oxidant, 0.2-3.0% of chlorine-containing reagent, 0.2-4% of acid-containing reagent, 0.1-2% of stabilizer, 1-3% of surfactant, 5-10% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 20-60% of cepharanthine, 10-40% of chlorogenic acid and 30-50% of tea saponin in percentage by mass.

Preferably, the oxidizing agent is one of an inorganic oxide or an organic oxide that can release oxygen ions. The inorganic oxide is selected from hydrogen peroxide and sodium peroxide, and the organic oxide is selected from peroxyacetic acid.

Preferably, the chlorine-containing agent is a hypochlorite-containing compound. The chlorine-containing agent is selected from sodium hypochlorite and hypochlorous acid.

Preferably, the acid-containing reagent is one or both of an inorganic acid or an organic acid. The inorganic acid is selected from phosphoric acid and hydrochloric acid, and the organic acid is selected from acetic acid, oxalic acid and citric acid.

Preferably, the surfactant is a cationic surfactant.

Preferably, the cationic surfactant is one of cetyltrimethylammonium bromide (CTAB) and cetyltriethylammonium bromide (CTEAB).

Preferably, the stabilizer is one of carbonate, borate and oxalic acid.

Oxidative and chlorine-containing agents disrupt the secondary and primary structure of DNA by oxidation.

The acid-containing reagent and stabilizer disrupt the phosphodiester and hydrogen bonds of the DNA by hydrolysis of hydrogen ions.

The cationic surfactant acts with hydrogen bonds of DNA through a hydrophobic thrust process to degrade the DNA.

Cepharanthine (CPA) is a monomer compound of dibenzylisoquinoline alkaloid, and is extracted and separated from tuber of plant of Stephania of Menispermaceae. Cepharanthine has wide biological activity, has been used clinically, is mainly used for treating leukopenia, and has no obvious toxic or side effect. Researches show that the cepharanthine also has an important effect in resisting the development of tumors, on one hand, the cepharanthine can promote the apoptosis of various malignant tumor cells including human leukemia cell lines, bile duct cancer cells, oral squamous cell carcinoma cells and the like; on the other hand, cepharanthine specifically destroys the activity of some viral ribonucleoprotein complexes.

Chlorogenic acid (Chlorogenic acid) is depside formed by the condensation of caffeic acid (caffeic acid) and quinic acid (quinic acid, 1-hydroxyhexahydrogallic acid), namely 3-O-caffeoylquinic acid (3-O-caffeoylquinic acid), and is a phenylpropanoid compound generated by plants through the shikimic acid pathway in the aerobic respiration process. Chlorogenic acid has wide bioactivity, and has antibacterial, antiviral, leukocyte increasing, liver protecting, gallbladder function promoting, antitumor, blood pressure lowering, blood lipid reducing, free radical scavenging, and central nervous system exciting effects.

The tea saponin is pure natural saponin extracted from seeds of Theaceae plants, and has the following properties: a white-like powder with a melting point of 224 c, is a surfactant. The tea saponin has emulsifying, dispersing, moistening and foaming properties, has pharmacological effects of anti-permeability, anti-inflammation, analgesia and the like, has the functions of sterilization, disinsection and stimulation of growth of certain plants, and is a main raw material for producing products such as an emulsifier, a detergent, a foaming agent, an insecticide and the like. More specifically, the tea saponin is a nonionic surfactant and a glycoside compound, can act with DNA together with a cationic surfactant, and can increase the degradation rate of the DNA under the synergistic action, and simultaneously realize the purpose of rapidly degrading the DNA under high-concentration DNA and low-concentration DNA.

Therefore, the nucleic acid pollution scavenger taking the traditional Chinese medicine extract as the main component, which adopts the structure, has the following beneficial effects:

1. the nucleic acid removing agent has mild and non-irritant odor, is simple and convenient to operate, can be directly sprayed on the floor, the wall surface, the table top of a laboratory, the table top of a biological safety cabinet, the surface of a liquid transfer device, the outer surfaces of various instruments, the outer wall of a centrifugal tube and the like during use, and can also be directly used for wiping the instruments, the table top and the like by the nucleic acid pollution removing agent.

2. The nucleic acid pollution scavenger disclosed by the invention is stable in property, convenient to store, long in storage period, safe and environment-friendly.

3. Through the multi-level degradation effect generated by the nucleic acid pollution scavenger and DNA, the cationic surfactant and the tea saponin cooperate to finally completely degrade nucleic acid fragments into nucleotides, the degradation effect of the nucleic acid can be completed within minutes, the generation of laboratory aerosol can be effectively avoided, and a good laboratory operation environment is provided.

4. The three traditional Chinese medicine extracts have the effects of resisting bacteria, sterilization and virus, can effectively destroy the structure of DNA to degrade the DNA and inhibit the synthesis of the DNA, and are low in toxicity, safe and environment-friendly.

The technical solution of the present invention is further described in detail by the following examples.

Detailed Description

The present invention will be further described below, and it should be noted that the present embodiment is based on the technical solution, and a detailed implementation manner and a specific operation process are provided, but the present invention is not limited to the present embodiment.

Example 1

A nucleic acid pollution scavenger taking traditional Chinese medicine extract as a main component comprises the following components in percentage by mass: the traditional Chinese medicine composition comprises 10% of a traditional Chinese medicine extract, 1.0% of hydrogen peroxide, 1.0% of sodium hypochlorite, 1.0% of phosphoric acid, 0.5% of oxalic acid, 1% of CTAB, 6% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 30% of cepharanthine, 30% of chlorogenic acid and 40% of tea saponin in percentage by mass.

A method for preparing nucleic acid pollution scavenger containing Chinese medicinal extract as main ingredient comprises the following steps:

(1) Weighing raw materials according to a proportion, and then mixing peroxide, a stabilizer, a chlorine-containing reagent, an acid-containing reagent and half of deionized water to obtain a first mixture;

(2) Mixing the other half of the deionized water with ethanol to obtain a mixture II;

(3) Mixing the mixture II with the traditional Chinese medicine extract to obtain a mixture III;

(4) And mixing and stirring the mixture III and the mixture I to obtain the nucleic acid pollution scavenger.

Example 2

A nucleic acid pollution scavenger taking traditional Chinese medicine extract as a main component comprises the following components in percentage by mass: the traditional Chinese medicine composition comprises 12% of a traditional Chinese medicine extract, 1.5% of sodium peroxide, 2.5% of hypochlorous acid, 3.0% of acetic acid, 1.5% of oxalic acid, 3% of CTEAB, 8% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 20% of cepharanthine, 30% of chlorogenic acid and 50% of tea saponin in percentage by mass.

The preparation method is the same as that of example 1.

Example 3

A nucleic acid pollution scavenger taking traditional Chinese medicine extract as a main component comprises the following components in percentage by mass: the traditional Chinese medicine composition comprises 13% of a traditional Chinese medicine extract, 0.8% of peroxyacetic acid, 0.6% of sodium hypochlorite, 3.5% of hydrochloric acid, 0.7% of oxalic acid, 1.4% of CTAB, 10% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 25% of cepharanthine, 32% of chlorogenic acid and 43% of tea saponin in percentage by mass.

The preparation method is the same as that of example 1.

Comparative example 1

Comparative example 1 is different from example 1 in that the herb extract is not added.

Comparative example 2

Comparative example 2 differs from example 1 in that tea saponin was not added.

Degradation of the different nucleic acid contamination scavengers prepared in examples 1-3 and comparative examples 1-2 was tested.

The nucleic acid contamination removing agents of the present examples and comparative examples were compared with various methods such as hydrogen peroxide and ultraviolet irradiation, and the effect of degrading nucleic acids by these treatment methods was observed. Among them, the nucleic acids to be compared were extracted plasmid pET32a, 50mg in one group, used at a normal concentration (i.e., directly synthesized concentration) with a nucleic acid contamination scavenger (examples 1-3 and comparative examples 1-2) for 3 minutes, 50mg in one group, irradiated with ultraviolet rays for 30 minutes at a distance of 0.5M and a wavelength of 254nm, and 50mg in one group, added with hydrogen peroxide to a final concentration of 0.01M for 5 minutes. The effect of nucleic acid degradation was judged by the presence of bright bands of varying degrees under the UV lamp.

It was found from the above experiments that examples 1-3 showed only faint traces of nucleic acid under irradiation of ultraviolet lamps, indicating that most of the nucleic acid was degraded, comparative examples 1-2 showed residual brightness under irradiation of ultraviolet lamps, indicating that only a part of the nucleic acid was degraded, and comparative example 1 had a slightly higher brightness than comparative example 2, and the ultraviolet light irradiation group and the hydrogen peroxide group showed brightness under irradiation of ultraviolet lamps, indicating that a large amount of nucleic acid was not degraded.

From the results of example 1 and comparative example 1, the effect of nucleic acid degradation is reduced without adding the traditional Chinese medicine extract, which shows that the traditional Chinese medicine extract plays a certain role in the nucleic acid degradation process and can realize the degradation of nucleic acid in a short time by cooperating with other substances.

From the results of example 1 and comparative example 2, it can be seen that the removal of tea saponin can also directly affect the effect of the nucleic acid pollution scavenger, and tea saponin on one hand serves as a traditional Chinese medicine extract to destroy the nucleic acid structure together with other two traditional Chinese medicine extracts through the actions of antibiosis, bacteriostasis and sterilization, and on the other hand serves as a nonionic surfactant to act on the groups on the DNA surface in cooperation with a cationic surfactant (CTAB) to degrade the DNA.

Therefore, the nucleic acid pollution scavenger taking the traditional Chinese medicine extract as the main component with the structure is adopted, through the multi-layer degradation effect generated by DNA, the cationic surfactant and the tea saponin are cooperated to finally degrade nucleic acid fragments into nucleotide completely, the degradation effect of the nucleic acid can be completed within a few minutes, the generation of laboratory aerosol can be effectively avoided, and a good laboratory operation environment is provided.

Finally, it should be noted that: the above embodiments are only for illustrating the technical solutions of the present invention and not for limiting the same, and although the present invention is described in detail with reference to the preferred embodiments, those of ordinary skill in the art should understand that: modifications and equivalents may be made to the invention without departing from the spirit and scope of the invention.

Claims (7)

1. A nucleic acid pollution scavenger taking a traditional Chinese medicine extract as a main component is characterized by comprising the following components in percentage by mass: 10-15% of traditional Chinese medicine extract, 0.4-2.0% of oxidant, 0.2-3.0% of chlorine-containing reagent, 0.2-4% of acid-containing reagent, 0.1-2% of stabilizer, 1-3% of surfactant, 5-10% of ethanol and the balance of deionized water, wherein the traditional Chinese medicine extract comprises 20-60% of cepharanthine, 10-40% of chlorogenic acid and 30-50% of tea saponin in percentage by mass.

2. The nucleic acid contamination scavenger based on Chinese medicinal extract as claimed in claim 1, wherein the oxidant is one of inorganic oxide or organic oxide capable of releasing oxygen ions.

3. The nucleic acid contamination removing agent comprising Chinese medicinal extract as main ingredient as claimed in claim 1, wherein the chlorine-containing agent is a compound containing hypochlorite.

4. The nucleic acid contamination scavenger based on Chinese medicinal extract as claimed in claim 1, wherein the acid-containing reagent is one or both of inorganic acid and organic acid.

5. The nucleic acid contamination scavenger based on Chinese medicine extract as claimed in claim 1, wherein the surfactant is cationic surfactant.

6. The nucleic acid contamination scavenger based on Chinese medicinal extract as claimed in claim 5, wherein the cationic surfactant is one of cetyl trimethyl ammonium bromide and cetyl triethyl ammonium bromide.

7. The nucleic acid contamination removing agent as claimed in claim 1, wherein the stabilizer is one of carbonate, borate and oxalic acid.