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Method for rapidly detecting polyoxin B using biological value detection and high performance liquid chromatography (HPLC)

A polyoxin and fast technology, applied in the field of medicine, can solve the problems of preparing many plates, heavy workload, and different antibacterial sizes, and achieve the effect of large plate area, reduced error, and large quantity

Inactive Publication Date: 2019-05-17
RUSHAN HANWEI BIO TECHN & SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] ③ The concentration of antibiotics is different, and the antibacterial size is also different. Compare the antibacterial diameter of the test sample and the standard product, and the content of the test sample can be obtained. Traditional bioassays generally use conventional 90mm and 120m petri dishes. There are many plates to prepare, and the workload is heavy, and each plate can hold up to 6 Oxford cups, and when drawing the standard curve, there are large errors because the standard solution is dropped in different petri dishes. Therefore, it is necessary to establish a set of intuitive , accurate, rapid and simple biological titer detection method has become an urgent problem to be solved

Method used

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  • Method for rapidly detecting polyoxin B using biological value detection and high performance liquid chromatography (HPLC)
  • Method for rapidly detecting polyoxin B using biological value detection and high performance liquid chromatography (HPLC)
  • Method for rapidly detecting polyoxin B using biological value detection and high performance liquid chromatography (HPLC)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Step 1. Determination of the biological potency determination conditions of polyoxin by cup and saucer method

[0022] Determination conditions: Dissolve polyoxin standard substance in phosphate buffer solution with pH=6.8, and then dilute to 25IU / mL, 50IU / mL, 100IU / mL in sequence. Estimate, dilute different times, and finally dilute to 50IU / mL according to the estimate, drop the standard sample and dilution solution into the Oxford cup in the same glass wooden tray, cover the glass and incubate for 16-20 hours, and measure the diameter of the inhibition zone , draw a standard curve, and calculate the sample content

[0023] ①Plate pouring: the upper medium is PDA medium, (200 grams of potatoes, peeled and cut into pieces, add 1000ml of distilled water to boil juice, filter with six layers of gauze, add to the scale, add 1% glucose, 2% agar powder, adjust PH =6.2), sub-packed into 120ml / 250ml Erlenmeyer flasks, sterilized for subsequent use;

[0024] 2% agar powder fo...

Embodiment 2

[0046] HPLC detection

[0047] The high-potency sample measured in Example 1 is detected by HPLC, and the comparison between the two is used to effectively reduce the workload and complete the accurate measurement of the content

[0048] HPLC detection conditions are as follows:

[0049] Chromatographic conditions: column brand Agilent, filled with Hypersil, C185um filler;

[0050] Specifications 250*10mm, detector wavelength 260nm, mobile phase methanol: water = 10:90 trifluoroacetic acid to adjust PH = 3.5-4.0, can effectively detect the chemical content of polyoxin.

[0051] Step 1. Linear relationship

[0052] Prepare a polyoxin standard solution with a certain concentration gradient, analyze according to the above method, repeat twice for each concentration, and take the average value of the peak area. With the mass concentration of polydactin as the abscissa and the peak area of ​​polydactin as the ordinate, draw a linear relationship diagram. The experimental result...

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Abstract

The invention relates to indicator bacteria using alternaria alternata as biological value detection. A high performance liquid chromatography (HPLC) chemical detecting method is combined, and the content of polyoxin B in samples can be rapidly and quantitatively detected. The detecting method uses self-made wooden discs as detecting tools, the number of the detected samples is nine times that ofthe a traditional method, corresponding standard curves correspond to the detecting glass wooden discs, the samples with high biological value can be rapidly and precisely screened, the chemical content of the samples with high content is detected by HPLC, the production cost is reduced, the labor intensity is reduced, and the method can be used for monitoring of the polyoxin B bacteria in the high-throughput screen and production processes.

Description

technical field [0001] The invention relates to a detection method in the technical field of medicines, in particular to the detection of the biological potency of polyoxin B and the detection of the HPLC chemical content. Background technique [0002] Polyoxin is a metabolite produced by fermentation of Streptomyces aureus. It is a broad-spectrum antibiotic fungicide. It is composed of homologues and isomers of A-N14 different components. It is a peptide pyrimidine nucleoside antibiotic , Mainly used to control apple leaf spot, ginseng black spot, rice sheath blight and other diseases. [0003] At present, the detection method of polyoxin B is mainly HPLC detection method, which is used for chemical analysis and measurement, but the HPLC method is expensive to detect, has a large flow consumption and strong toxicity, and the pre-treatment steps are complicated and the analysis time is long, which is not only suitable for large-scale It is used in strain screening and produ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N30/02G01N30/74
Inventor 徐晓庆杨孟森宫照普
Owner RUSHAN HANWEI BIO TECHN & SCI
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