A method for screening nifedipine by high performance thin layer chromatography combined with bioluminescence

A technology of high-performance thin-layer chromatography and nifedipine, which is applied in the field of high-performance thin-layer chromatography combined with bioluminescence to screen nifedipine, and can solve problems such as background interference and lack of selectivity for multiple targets

Active Publication Date: 2020-06-16
JIANGNAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this mode is simple and efficient, it is not suitable for the detection of illegal addition of Chinese and Western medicines in tea, because it has the following defects: ①Serious background interference
② Lack of selectivity for multiple targets

Method used

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  • A method for screening nifedipine by high performance thin layer chromatography combined with bioluminescence
  • A method for screening nifedipine by high performance thin layer chromatography combined with bioluminescence
  • A method for screening nifedipine by high performance thin layer chromatography combined with bioluminescence

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Preparation of working luminescence suspension by high performance thin layer chromatography coupled with bioluminescence

[0032] The preparation process of the working luminescent suspension is as follows:

[0033] (1) Preparation of simulated seawater liquid:

[0034] Configure the simulated seawater liquid medium according to the following formula: 30g / L NaCl, 5g / L Na 2 HPO 4 , 5g / LKH 2 PO 4 , 3mL / L glycerin, 5g / L peptone and 5g / L yeast extract; after adding 1L ultrapure water and stirring to dissolve, adjust the pH value of the prepared liquid medium to 7 with 1mol / L sodium hydroxide aqueous solution, and then Use a high-pressure steam sterilizer to sterilize at 121°C for 15 minutes. The prepared liquid medium is packaged and stored in the refrigerator for later use. When not in use, it can be stored at 4°C for 7 days;

[0035] (2) Cultivation and preservation of luminescent bacteria: Inoculate the luminescent bacteria cryopreserved with glycerol in...

Embodiment 2

[0036] Example 2: Drawing of a standard curve based on high performance thin-layer chromatography coupled with bioluminescence

[0037](1) Standard solution preparation: Accurately weigh 50 mg of nifedipine standard substance with an electronic balance, place it in a 10mL volumetric flask, and constant volume with methanol to obtain a standard stock solution with a concentration of 5mg / mL; keep the standard stock solution away from light at ordinary times Put it in an environment of 4°C, and when the analysis is about to start, take out 1mL and place it in a 10mL volumetric flask, dilute it with methanol to obtain a standard working solution with a concentration of 0.5mg / mL; the standard working solution must be reconfigured before each test;

[0038] (2) The prepared standard solution is directly used for HPTLC spotting;

[0039] (3) Chromatographic separation: first, manually draw the solution to be tested with a 100 μL sampling needle, and with the assistance of a 0.5 MPa n...

Embodiment 3

[0044] Example 3: Condition optimization of high performance thin layer chromatography coupled with bioluminescence

[0045] 1. The influence of different silica gel plates on the test results

[0046] The detection principle of the method is: the luminescent bacterial strain is specifically a type of microorganism capable of emitting visible light under normal physiological conditions. The luminescent principle of this type of bacteria is that under the action of luciferase catalysis and molecular oxygen, long-chain fatty aldehydes and reduced flavin mononucleotides (FMNH2) are oxidized to long-chain fatty acids and oxidized flavin mononucleotides (FMN), At the same time, blue-green light with a wavelength of 450-490nm is released. Based on this characteristic, luminescent bacteria are often used as optical sensing elements of biodetectors. Compared with common physical and chemical detection methods, the biggest advantage of luminescent bacterial biosensing lies in the non...

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Abstract

The invention discloses a method for screening nifedipine by a high-performance thin-layer chromatography combined with a bioluminescent method, and belongs to the technical field of food detection. The high-performance thin-layer chromatography and the bioluminescent method are combined in the invention; a sample to be detected is spread on a thin-layer plate at first; then, the sample to be detected is coupled to luminous bacteria; luminous imaging is carried out; and finally, thin-layer chromatography scanning is carried out. The method in the invention can detect multiple samples, and candetect 20 samples at most for one time; the single experimental duration is 30 min; only one sample can be detected for one time by adoption of a HPLC method; the single experimental duration is 20 min; compared with the existing high-performance liquid chromatography, the method in the invention can realize rapid screening; furthermore, multiple samples can be simultaneously detected in a singletest; high-throughput screening can be realized; the recovery rate of the method is 96.8%-98.7%; the detection limit can be up to 0.5-1 mug / kg; the detection method can realize that the repeatabilityRSD is less than 2.3%; and the method has the advantages of being economic, rapid, simple and convenient.

Description

technical field [0001] The invention relates to a method for screening nifedipine by high-efficiency thin-layer chromatography combined with bioluminescence, and belongs to the technical field of food detection. Background technique [0002] Nifedipine (Nifedipine, NDP) belongs to the antihypertensive chemical drugs of dihydropyridines. Its pharmacodynamic principle is that it is a calcium antagonist, which can selectively inhibit the transmembrane transport of calcium ions into cardiomyocytes and smooth muscle cells, and inhibit Calcium ions are released from the cells without changing the plasma calcium ion concentration, thereby achieving the effect of lowering blood pressure. Nifedipine-like blood pressure-lowering chemical drugs have good efficacy and less side effects, and are easily used to adulterate health teas. In the health care product market, blood pressure lowering health care products derived from medicinal and edible materials occupy a large share. Accordin...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/90G01N30/95G01N21/76
CPCG01N21/763G01N30/90G01N30/95
Inventor 陈益胜黄彩虹徐学明舒蓝萍金征宇谢正军田耀旗杨哪金亚美龙杰詹征峰周宇计海燕
Owner JIANGNAN UNIV
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