Application of p-toluidine blue_Kain Industrial Additive

Background and overview^[1][2]

P-Toluidine blue is an organic reagent, a topical drug used to increase the positive rate of vulvar cancer biopsy. Apply 1% p-toluidine blue solution to the vulva, let it dry for 2 to 3 minutes, and then use 1% acetic acid to decolorize it. If there is atypical hyperplasia, the lesion area of ​​carcinoma in situ or invasive cancer will appear purple-blue without decolorization. Biopsy in non-depigmented areas can increase the positive rate of early diagnosis and diagnosis of multiple central cancers. However, this method can cause false positive results for benign ulcers; and for atypical lesions with hyperkeratosis on the surface, it can be depigmented after washing with acetic acid and give false negative results.

Pharmacological effects^[2]

Wood is an important part of my country’s sustainable development strategy. Wood decay is one of the main ways to destroy wood quality. White rot fungi are the most serious damage to wood cell walls among various wood-rot fungi. In order to inhibit pathogenic bacteria and improve wood utilization, Yu Hongfeng and others used low-toxicity and low-pollution p-toluidine blue (TB)-mediated photodynamic therapy to study the inhibitory effect of white rot fungi. When different concentrations of TB (0. 01, 0. 02, 0. 05 mg/mL) were tested on white rot bacteria by the Oxford cup method, no antibacterial effect was observed. When using low-power 630nm laser to excite TB, it was found that three groups of photosensitizer TB had an inhibitory effect on white rot bacteria. As the TB concentration increased, the inhibitory rates were 69.67%, 91.21%, and 99.44% respectively. Experimental results show that p-toluidine blue-mediated photodynamic therapy has a good antibacterial effect on white rot bacteria, and the photosensitizer concentration is positively correlated with its antibacterial effect, reflecting a good wood preservative effect.

Apply^[3-5]

1. CN201310599323.2 provides a p-toluidine blue-DNAse agar medium. The components of the medium include 15.5g/L 7.5% sodium chloride broth, 0.5g/L DNA, 1.5g /L calcium chloride, 6.3g/L trishydroxymethylaminomethane, 0.088g/L p-toluidine blue, pH value is 8.9-9.1.

The application of the toluidine blue-DNAse agar medium is used for ribonuclease detection.

Bacteria with deoxyribonuclease decompose deoxyribonucleic acid (DNA) in the culture medium, hydrolyzing long DNA chains into short oligonucleotide chains of several single nucleotides, causing the indicator to react with toluidine blue Turns pink; calcium chloride provides cations as an activator; sodium chloride maintains a balanced osmotic pressure; trishydroxymethylaminomethane is a buffer; agar is the coagulant of the culture medium.

Weigh 27.58g of this product, heat and dissolve it in 1000ml distilled water, and put it into Erlenmeyer flasks.

Sterilization is not required if used immediately. If not used immediately, autoclave at 121°C for 15 minutes.

Sterilized media can be stored unchanged for 4 months at room temperature and can still be used after being thawed several times.

Beneficial effects of the present invention:

1. The medium formulation is simple, the preparation operation is convenient, the cost is low, and it is conducive to popularization and application;

2. Bacteria grow rapidly and have low mutation rate;

3. Easy to observe and analyze.

2. CN201110054955.1 discloses a combined reagent for screening early lesions of oral lining cells, including: p-toluidine blue as a detection reagent and biochemical reagent, and acetic acid as an auxiliary reagent for the detection reagent. The p-toluidine blue The concentration of the solution is 0.2~2%, and the concentration of the acetic acid solution is 0.01~5%. The invention also discloses a method for using a combined reagent for screening early lesions of oral endothelial cells, which is characterized by: first gargling with acetic acid for 1-2 minutes, and then gargling with a mixed solution of p-toluidine blue and acetic acid for 1-2 minutes. minutes, and finally rinse your mouth with acetic acid for 1-2 minutes. The ratio of the p-toluidine blue and acetic acid mixed solution is: 1:0.5-1.

3. CN201610342904.1 provides a preparation method and application of a mycotoxin immunosensor based on graphyne-toluidine blue composite material

(1) Use Al₂O₃ polishing powder to polish the glassy carbon electrode with a diameter of 4 mm smoothly, and clean it with ultrapure water; remove 5 ~ 8 µL, 0.4 ~ 4 mg/mL graphdiyne@p-toluidine blue Gy@TB composite material was dropped onto the electrode surface, rinsed with ultrapure water, and dried at room temperature to form a film;

(2) Drop 3 ~ 8 µL with a mass fraction of 0.1% ~ 5% hydroxypropyl chitosan on the electrode surface, rinse with ultrapure water, and dry at room temperature;

(3) Add 1 ~ 5 µL dropwise, with a mass fraction of 0.1% ~ 1% pentanePut the dialdehyde aqueous solution on the electrode surface, rinse it with ultrapure water, and dry it at room temperature;

(4) Add 5 ~ 8 µLmycotoxin antibody, 5 ~ 8 µL, and bovine serum albumin BSA with a mass fraction of 0.5% ~ 3% in sequence. Apply the solution to the electrode surface, rinse with ultrapure water, and dry in a refrigerator at 4°C;

(5) Drop 5 ~ 8 µL, 0.06 ~ 40 ng/mL of a series of different concentrations of mycotoxin antigen standard solutions onto the electrode surface, rinse with ultrapure water, and store in a 4°C refrigerator Leave to dry.

Preparation of graphyne@p-toluidine blue Gy@TB composite material: add 1 ~ 10 mg of graphyne Gy and 0.5 ~ 5 mg of p-toluidine blue TB into 0.5 ~ 5 mL of ultrapure water, at room temperature After shaking for 24 h, the composite material graphyne@p-toluidine blue Gy@TB was obtained.

Preparation method and application of a mycotoxin immunosensor based on graphyne-toluidine blue composite material. The mycotoxin is selected from one of the following: DON, zearalenone, fumonisins , aflatoxin B1, ochratoxin.

Beneficial results of the present invention

(1) The use of graphdiyne has successfully enlarged the specific surface area of ​​the bottom layer of the sensor, and at the same time can well promote electron transfer on the electrode surface.

(2) The use of graphdiyne@p-toluidine blue Gy@TB composite material increases the number of signal molecules, expands the linear range of the sensor, and lowers the detection limit.

(3) The use of hydroxypropyl chitosan enhances the firm fixation of the Gy@TB composite material on the sensor surface, and at the same time realizes the chemical bond connection to the antibody, achieving ultra-sensitive and stable detection.

Main reference materials

[1] Chinese Sexual Medicine Dictionary

[2] Yu Hongfeng, Mou Hongbo, Qi Dawei, Yu Ying. Inhibition of a white rot fungus by p-toluidine blue-mediated photodynamic therapy [J]. Forest Engineering, 2019, 35(02): 50-54+81.

[3] CN201310599323.2 A kind of p-toluidine blue-DNAse agar medium and its application

[4] CN201110054955.1 Combination reagents for screening early lesions of oral lining cells and methods of use

[5]CN201610342904.1 Preparation method and application of mycotoxin immunosensor based on graphyne-toluidine blue composite material