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Liquid chromatography-tandem mass spectrometry detection method for emodin and aloe-emodin in mouse urine

A technology of aloe-emodin and liquid chromatography, which is applied in the detection of urine metabolites, liquid chromatography-tandem mass spectrometry detection of emodin and aloe-emodin in mouse urine, can solve the problems of complex steps and insufficient sensitivity, Achieve the effect of simple operation, high sensitivity, and improve detection sensitivity

Inactive Publication Date: 2020-11-06
CHINA TOBACCO JIANGXI IND CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current detection method of emodin and aloe-emodin in urine is commonly used liquid chromatography, the detection sensitivity is not high enough, and the solvent needs to be evaporated after the urine sample is extracted, and the sample is injected after concentration. The steps are relatively complicated

Method used

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  • Liquid chromatography-tandem mass spectrometry detection method for emodin and aloe-emodin in mouse urine
  • Liquid chromatography-tandem mass spectrometry detection method for emodin and aloe-emodin in mouse urine
  • Liquid chromatography-tandem mass spectrometry detection method for emodin and aloe-emodin in mouse urine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Pretreatment method

[0029] Take the urine of the rats fed with the 3# material solution and thaw it at room temperature, add 200 μL of the thawed sample, add 800 μL deionized water, 100 μL mixed internal standard solution, filter through a 0.22 μm organic phase filter, and perform HPLC-MS / MS analysis.

[0030] standard curve drawing

[0031] 1) Configuration of standard stock solution

[0032] Accurately weigh 10mg of emodin and 17.8mg of aloe-emodin into 100mL brown volumetric flasks, dilute to the mark with methanol, store the concentration of the standard stock solution in the dark at -18°C, and store it in the dark. The validity period is 3 months, see the table 1.

[0033] Table 1 feed liquid composition standard stock solution

[0034] standard stock solution Concentration (μg / mL) Emodin 100 Aloe-emodin 178

[0035] 2) Configuration of mixed standard solution

[0036] Pipette 1.0 mL each of emodin standard stock solution and al...

experiment example 2

[0074] The detection of experimental example 2 samples

[0075] The established analysis method was used to analyze and detect the composition of urine feed liquid of 6 rats (blank control), 6 rats of 3# feed liquid and 6 rats of 4# feed liquid gavaged for 24 hours. Analysis and detection obtained two metabolites of feed liquid: aloe-emodin and emodin. The metabolic components showed the same rule: the control group<3# feed liquid<4# feed liquid, the analysis results are shown in Table 10.

[0076] Contents of emodin and aloe-emodin in the urine of table 10 gavage rats

[0077]

[0078] The invention selects 0.1% formic acid aqueous solution and acetonitrile as the mobile phase, selects ultra-high performance liquid phase and tandem mass spectrometry, gradient elution, the separation degree of emodin and aloe-emodin meets the requirements, and the system adaptability is good.

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Abstract

The invention discloses a liquid chromatography-tandem mass spectrometry detection method for emodin and aloe-emodin in mouse urine, which comprises the following steps of S1, unfreezing the collectedurine, and recovering to room temperature, S2, taking the urine obtained in the step 1, and adding deionized water for mixing, S3, adding an internal standard into the mixed solution, S4, filtering the mixed solution, and S5, taking the filtered mixed solution for determination by a liquid chromatography-mass spectrometer, the liquid chromatography mobile phase being a water-acetonitrile system,the liquid chromatography column being a C18 reversed-phase liquid chromatography column, and the flow velocity of the liquid chromatography column being not greater than 0.5 mL / min. The method has the advantages of high sensitivity, simple operation, high stability and reliable result.

Description

technical field [0001] The invention relates to a method for detecting urine metabolites in the technical field of bioengineering, in particular to a method for detecting emodin and aloe-emodin in mouse urine by liquid chromatography tandem mass spectrometry. Background technique [0002] Rhubarb medicinal materials are derived from the dried roots and rhizomes of plants belonging to the genus Rheum in the family Polygonaceae (Polgynoaceae). It has the effects of venting, antibacterial, anti-tumor, anti-hyperlipidemia, lowering blood pressure, invigorating the stomach, promoting gallbladder, protecting the liver, strengthening the heart, anti-inflammatory, delaying aging, and regulating immunity. Rhubarb is one of the four most commonly used traditional Chinese medicines in my country. More than 800 of the more than 8,000 traditional Chinese medicine preparations in the country contain rhubarb. It is an important traditional Chinese medicine for treating various diseases and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/14G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/14G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 蔡继宝苏加坤郭磊徐达罗娟敏尚平平李翔谢复炜
Owner CHINA TOBACCO JIANGXI IND CO LTD
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