Method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry

A technology of liquid chromatography and mass spectrometry, applied in the field of drug analysis, to achieve the effect of high accuracy and good reproducibility

Active Publication Date: 2019-01-15
山东国仓健生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

There is no related report on simultaneous detection of tomatine and tomato saponin

Method used

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  • Method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry
  • Method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry
  • Method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1: Determination of tomatine and tomato saponin by liquid chromatography-mass spectrometry

[0032] 1. Sample pretreatment:

[0033] (1) The tomato material is ground into powder by adding liquid nitrogen, and 40 mg of the material is weighed in a centrifuge tube.

[0034] (2) Add 1.5ml of pure methanol to the sample, and vortex for 10s.

[0035] (3) Centrifuge at 3000rpm for 10min at 4°C.

[0036] (4) Take 100 μl of the 1.5ml supernatant, pass it through a 0.22 micron PVDF filter membrane, and put it into a glass sample injection vial with an inner insert.

[0037] 2. Preparation of standard working solution:

[0038] Tomatine standard substance (purchased from Shanghai Yuanye Biotechnology Co., Ltd., purity>95%) was dissolved in methanol to prepare standard samples with concentrations of 0.1ng / μl, 1.0ng / μl, and 2ng / μl; tomato saponin Standards (purchased from TRCI) were dissolved in methanol to prepare standard samples with concentrations of 0.1 ng / μl, 1.0...

Embodiment 2

[0057] Example 2: Methodological Validation

[0058] 1. Recovery test:

[0059] (1) Take 8 parts of 40mg tomato material and add pure methanol extract.

[0060] (2) Add 400ng tomatine standard substance and 100ng tomato saponin standard substance to 4 of the materials respectively, and mark them. No standard samples were added to the remaining 4 materials.

[0061] (3) Operate according to the extraction process of Example 1, inject samples, and run samples.

[0062] (4) Data processing, calculate the recovery rate according to the result.

[0063] Recovery rate experiment results:

[0064] The recovery rate of tomatine: 77.51%; the recovery rate of tomato saponin: 91.66%.

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Abstract

The invention discloses a method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry. The method comprises the following steps of: (1) preprocessinga sample to be tested; (2) measuring the content of tomatine and tomato saponin in the preprocessed sample by liquid chromatography-mass spectrometry. The method for measuring tomatidine and tomato saponin by high performance liquid chromatography-mass spectrometry realizes simultaneous detection of tomatidine and tomato saponin in tomato sample materials. The method meets the requirement of quantitative analysis in precision, accuracy, and stability. The method has the characteristics of high accuracy, good reproducibility, stability, and reliability. The method can also be used for quantitative analysis of changes in tomatine content during tomato breeding.

Description

technical field [0001] The invention relates to the technical field of drug analysis, in particular to a method for determining tomatine and tomato saponin by liquid chromatography-mass spectrometry. Background technique [0002] Tomatine, also known as tomatine, is a secondary metabolite produced in tomato rhizomes, leaves, and green fruits, and has the highest content in tomato plants. Tomatine can protect tomato plants from bacteria, fungi, viruses and certain insects during the growth process, and has obvious antibacterial and insect repellent effects. According to literature reports, tomatine can inhibit certain pathogenic bacteria harmful to humans and animals, and has significant biological activity. [0003] Tomato saponin is a water-soluble saponin compound isolated from tomato. It mainly exists in the fruit of tomato, and also has a considerable amount in seeds, leaves, stems and other parts. Tomato saponins have the properties of general saponins. Foreign resear...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06
Inventor 胥倩路雅茹谷淑波李彩霞
Owner 山东国仓健生物科技有限公司
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