Aristolochic acid fluorescent test paper and preparation method and application thereof

A technique for aristolochic acid and test paper, applied in the field of fluorescent test paper, can solve the problem of increased confusion, misselling, mispurchase and misuse of aristolochic acid medicinal materials, purchase and maintenance costs of complex pretreatment instruments, and large time consumption, etc. problem, to achieve good detection sensitivity, low cost and obvious effect

Active Publication Date: 2018-12-21
宁波鉴合科技有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

At present, the methods for analyzing and detecting aristolochic acid are mainly liquid chromatography and liquid chromatography-mass spectrometry, and these methods require professionals to consume a lot of time to operate, and require complex pretreatment and expensive instrument purchase. Expenses and maintenance costs, these cost

Method used

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  • Aristolochic acid fluorescent test paper and preparation method and application thereof
  • Aristolochic acid fluorescent test paper and preparation method and application thereof
  • Aristolochic acid fluorescent test paper and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Embodiment 1: the preparation of PPE-PEG fluorescent test paper.

[0032] Compound M 1 Synthesis:

[0033] In a three-neck flask, disperse potassium hydroxide powder (2.24g, 40mmol / L) in 20mL of DMSO and stir vigorously, add hydroquinone (0.88g, 8mmol) quickly under an argon atmosphere, and then dissolve 4-bromo Ethyl butyrate (4.6mL, 32mmol) was slowly injected into the above mixture, and the reaction mixture was stirred for 48h, extracted with dichloromethane, washed with saturated brine, and finally washed with anhydrous Na 2 SO 4 After drying, the solvent was distilled off under reduced pressure to obtain a crude product, and the crude product was purified by column chromatography (ethyl acetate / petroleum ether, volume ratio 1:5), and then the solvent was distilled off under reduced pressure to obtain the product white powder 1,4-bis(4 -Oxybutyrate ethyl)benzene (M 1 ).

[0034] Compound M 2 Synthesis:

[0035] KI (0.31g, 1.44mmol) and I 2 (0.8g, 3.15mmol) w...

Embodiment 2

[0042] Embodiment 2: the measurement of fluorescent probe fluorescent property

[0043] Take the 22.4mg PPE-PEG powder prepared in Example 1 and dissolve it with PBS buffer solution (0.05mol / L, pH=7.2) and settle it into a 500mL volumetric flask to prepare PPE-PEG with a monomer concentration of 0.5mmol / L Stock solution, protected from light. Take 30 μL from the stock solution and add it to a 5 mL centrifuge tube, add 30 μL aristolochic acid standard solution (10 mmol / L) and dilute to 3 mL with PBS buffer solution (50 mmol / L, pH=7.2), then measure its fluorescence properties. Depend on figure 2 It can be seen that the fluorescence maximum excitation wavelength of the aristolochic acid fluorescent probe is 370nm, and the maximum emission wavelength is 435nm.

[0044] Pipette several parts of 30 μL of the PPE-PEG stock solution (0.5mmol / L) prepared in the above steps, add 30 μL of various concentrations of aristolochic acid solutions to it, so that the concentration of aristo...

Embodiment 3

[0045] Embodiment 3: The detection of PPE-PEG fluorescent probe to actual sample

[0046] In order to verify the detection effect of the fluorescent probe PPE-PEG on the content of aristolochic acid in several Chinese patent medicines on the market, several Chinese patent medicines containing aristolochic acid were purchased in the pharmacy and processed according to the literature A simple pretreatment was carried out. After grinding the Chinese patent medicine with a mortar, accurately weigh 5g of the powder and put it into a 50mL centrifuge tube, add 20ml of ethanol for ultrasonic extraction for 24h, then centrifuge the mixed solution to take the supernatant and remove the drug residue Wash with ethanol three times, combine the ethanol part and extract the ethanol phase with petroleum ether for 5 times, then spin dry the separated ethanol phase, dissolve the precipitated solid with 10 mL of chloroform, filter out the insoluble matter, and dilute the organic phase to 25 mL wi...

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Abstract

The invention discloses an aristolochic acid fluorescent test paper and a preparation method and application thereof, and belongs to the technical field of the medicine analysis. The preparation method comprises the following steps: firstly synthesizing 1,4-di(4-oxybutyrate) benzene, and then iodizing the benzene ring by using KIO3 and I2, polymerizing with the 1,4-diethylbenzene under the catalysis of the tetrakis(triphenylphosphine)palladium to obtain polymer precursor PPE-OBA; synthesizing the oxybutyric acid as oxybutyrate polyethylene glycol through esterification reaction on the PPE-OBAside chain, and obtaining an aristolochic acid fluorescent probe PPE-PEG, wherein the structure is as shown in description; finally soaking the filter paper strip in a PPE-PEG solution for 30sec, placing in a dark room for 1 hour after taking out from the PPE-PEG solution, thereby obtaining the aristolochic acid fluorescent test paper. The fluorescent test paper disclosed by the invention is convenient for preparation, low in cost, obvious in effect and has the potential for large-scale manufacturing; the aristolochic acid can be quickly detected on the spot in real time by naked eyes by onlyneeding a handheld ultraviolet lamp, and the test paper has extensive application prospect.

Description

technical field [0001] The invention belongs to the technical field of drug analysis, and relates to a fluorescent test paper capable of visually and quickly detecting aristolochic acid in a drug sample, a production method and an application thereof. Background technique [0002] Aristolochic acid (AA), also known as total aristolochic acid, phagocytic acid or aketoxin, is a class of nitrophenanthrene carboxylic acids. Aristolochic acid exists in Aristolochiaceae plants such as Aristolochia and Asarum. These plants have been widely used as Chinese herbal medicine to treat a variety of common diseases, but at the same time, aristolochic acid is a substance with nephrotoxicity. Nephropathy caused by aristolochic acid has been reported in France, Germany, Spain, the United Kingdom, the United States, Both Japan and China have reported. On October 27, 2017, in the list of carcinogens released by the International Agency for Research on Cancer of the World Health Organization,...

Claims

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Application Information

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IPC IPC(8): G01N21/64
CPCG01N21/6447
Inventor 冯国栋刘吉林
Owner 宁波鉴合科技有限责任公司
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