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Detection method of echinocandin

A detection method, echinocandin technology, applied in the field of medicine, can solve problems such as low efficiency, column efficiency and stability to be improved

Pending Publication Date: 2020-07-07
LUNAN PHARMA GROUP CORPORATION
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Currently, for echinocandin B 0 selective detection of echinocandin C 0 It is inefficient in terms of the selective detection of the two, and the column efficiency and stability of the simultaneous detection of the two need to be improved

Method used

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  • Detection method of echinocandin
  • Detection method of echinocandin
  • Detection method of echinocandin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0072] Detection conditions:

[0073] Column: HILIC Amide, 4.6*250mm, filler particle size 3.5μm;

[0074] Mobile phase: A phase is water, B phase is acetonitrile, gradient elution;

[0075] The column temperature is 40°C;

[0076] The flow rate is 1.0mL / min;

[0077] Injection volume 10μL;

[0078] Detection wavelength 220nm;

[0079] During gradient elution, the elution gradients of mobile phase A and B are as follows:

[0080]

[0081] Configure the test solution:

[0082] Measure 20ml of echinocandin fermentation liquid in a tank, add 80ml of absolute ethanol (AR) to homogenate for 10min, filter, take an appropriate amount of filtrate and centrifuge at 4000r / min for 10min, take the supernatant and filter it with a 0.45μm filter membrane to obtain The test solution.

[0083] Configuration B 0 and C 0 The reference substance solution:

[0084] B 0 Reference solution

[0085] Precise weighing of echinocandin B 0 Put 10.00 mg of the reference substance in a ...

Embodiment 2

[0092] Detection conditions:

[0093] Column: HILIC Amide, 4.6*250mm, filler particle size 3.5μm;

[0094] Mobile phase: A phase is water, B phase is acetonitrile, gradient elution;

[0095] The column temperature is 30°C;

[0096] The flow rate is 1.5mL / min;

[0097] Injection volume 20μL;

[0098] Detection wavelength 200nm;

[0099] During gradient elution, the elution gradients of mobile phase A and B are as follows:

[0100]

[0101] Need testing solution and reference substance solution are prepared as described in embodiment 1;

[0102] Measure the B of the above-mentioned reference substance solution and need testing solution by this detection condition 0 and C 0 , record its retention time, number of theoretical plates and resolution, and the test results are shown in Table 6.

[0103] Table 6 Test results

[0104]

Embodiment 3

[0106] Detection conditions:

[0107] Column: HILIC Amide, 4.6*250mm, filler particle size 3.5μm;

[0108] Mobile phase: A phase is water, B phase is acetonitrile, gradient elution;

[0109] The column temperature is 50°C;

[0110] The flow rate is 2.5mL / min;

[0111] Injection volume 15μL;

[0112] Detection wavelength 230nm;

[0113] During gradient elution, the elution gradients of mobile phase A and B are as follows:

[0114]

[0115] Need testing solution and reference substance solution are prepared as described in embodiment 1;

[0116] Measure the B of the above-mentioned reference substance solution and need testing solution by this detection condition 0 and C 0 , record its retention time, number of theoretical plates and resolution, and the test results are shown in Table 7.

[0117] Table 7 Test results

[0118]

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Abstract

The invention belongs to the technical field of medicines, and provides a high performance liquid chromatography method for detecting echinocandin B0 and C0, wherein the detection conditions are as follows: a chromatographic column is Ultra Amide, 4.6 * 250 mm, the filler particle size is 3.5 [mu]m, the column temperature is 30-50 DEG C, the flow rate is 1-2.5 mL / min, the sample injection volume is 10-20 microlitres, the detection wavelength is 200-230 nm, and a mobile phase adopts water and acetonitrile for gradient elution. The detection method is high in precision, high in accuracy, good inreproducibility, simple, convenient and practical, can realize simultaneous detection of echinocandin B0 and echinocandin C0, and has a very wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to a high performance liquid chromatography detection method of echinocandins. Background technique [0002] Since the 1970s, in the course of clinical treatment, fungal infection diseases and the death rate caused by uncontrollable fungal infections have gradually increased, which is related to the widespread use of drugs that damage the human immune system and the extensive use of broad-spectrum antibacterial drugs. direct relationship. Also, associated with clinical use of internal implants and infection with chronic immunosuppressive viruses like AIDS. Therefore, it is very important to research and develop safe, new and effective antifungal drugs. As we all know, fungal cells have cell walls, mammalian cells do not have cell walls. Therefore, the cell wall of fungi should be used as the target of new antifungal drugs. Echinocandins are drugs that act on the cel...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 张贵民魏荣省李旭娇
Owner LUNAN PHARMA GROUP CORPORATION
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