Method used for separating and identifying flavonoid matters in tobacco by adopting liquid chromatography-mass spectrography

A liquid chromatography and identification method technology, applied in the field of plant extract separation and identification, can solve problems such as limited sensitivity of ultraviolet detectors, achieve simple and rapid sample preparation, improve detection efficiency, and good accuracy

Active Publication Date: 2014-10-15
YUNNAN ACAD OF TOBACCO AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limited sensitivity of the ultraviolet detector, it can only be used for the analysis and detection of flavonoid substances with high content, and many trace or even trace flavonoid substances cannot be found by ultraviolet detectors

Method used

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  • Method used for separating and identifying flavonoid matters in tobacco by adopting liquid chromatography-mass spectrography
  • Method used for separating and identifying flavonoid matters in tobacco by adopting liquid chromatography-mass spectrography
  • Method used for separating and identifying flavonoid matters in tobacco by adopting liquid chromatography-mass spectrography

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] ——Isolation and Identification of Isorhamnetin in Tobacco

[0045] (1) Experimental reagents and devices

[0046] Acetonitrile (chromatographic grade), methanol (chromatographic grade), and ethanol (chromatographic grade) were purchased from Merck, Germany. Ultrapure water was produced by a Millipore purification system. Other standard samples were purchased from Sigma-Aldrich, Alfa Aesar and Bailingwei. Waters UPLC liquid chromatograph (USA), AB SCIEX 5500 triple quadrupole mass spectrometer (USA). SB-50D ultrasonic extractor (Ningbo Xinzhi Biotechnology Co., Ltd.), Waters BEH C18 (15 cm×2.1 mm×1.7 μm) reversed-phase chromatographic column (Waters, USA), MILLI-Q pure water machine (MILLIPORE) , LD5-2A centrifuge (Beijing Jingli Centrifuge Co., Ltd.), vortex mixer (Breda, Netherlands), CP2245 analytical balance (sensitivity 0.0001g, Sartorius, Germany).

[0047] (2) Experimental materials

[0048] The mature middle leaves and petals of Yunyan 97, Honghua Dajinyuan,...

Embodiment 2

[0056] ——Isolation and identification of isorhamnetin-3-O-rutinoside from tobacco

[0057] (1) Experimental reagents and devices

[0058] With embodiment 1.

[0059] (2) Experimental materials

[0060] With embodiment 1.

[0061] (3) Experimental method

[0062] With embodiment 1.

[0063] (4) Experimental results

[0064] Since isorhamnetin was found in tobacco samples, it was hypothesized that glycosyl-bound compounds of isorhamnetin also exist in tobacco. The molecular ion of isorhamnetin (315 - ) was used as a characteristic ion to perform a full scan of the parent ion, and it was found that there was an obvious chromatographic peak at the retention time of 4.55 min ( Figure 4 ), the detected precursor ion is 623 - . According to 623 - with 315 - There is a neutral loss of 308, and the compound is preliminarily judged to be isorhamnetin-rutinoside. Collect 4.55 min at 623 - Ion secondary fragment ion, and submit it to the mass bank website (www.massbank.jp) f...

Embodiment 3

[0066] ——Isolation and Identification of Quercetin-3-O-Rutinoside from Tobacco

[0067] (1) Experimental reagents and devices

[0068] With embodiment 1.

[0069] (2) Experimental materials

[0070] With embodiment 1.

[0071] (3) Experimental method

[0072] With embodiment 1.

[0073] (4) Experimental results

[0074] at 151 - When scanning precursor ions for characteristic ions, there is an obvious chromatographic peak at the chromatographic retention time of 5.80 min, and the detected precursor ion is 301 - . Collect 301 at 5.80 min -Ion secondary fragment ion, and submit it to the mass bank website (www.massbank.jp) for spectral library comparison, and found that it is very similar to the secondary mass spectrum of quercetin in the spectral library, buy quercetin The standard substance of the compound was compared with the retention time and the secondary mass spectrum of the compound to be identified, and it was confirmed that it was quercetin. Molecular...

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Abstract

The invention discloses a method used for separating and identifying flavonoid matters in tobacco by adopting liquid chromatography-mass spectrography. The method comprises the following steps: a sample is ground and dried, an extractant is added according to the standard that 0.1 ml of the extractant is added into 1 mg of the sample for extraction and centrifugation, a supernatant liquor is collected, water and chloroform are added according to the standard that 0.5 ml of the water and 1 ml of the chloroform are added into 1 ml of the supernatant liquor for centrifugation, and the supernatant liquor is collected for analysis; 153+ and 151- are adopted as characteristic ions for scanning, parent ions are extracted for second level mass spectrum scanning, and an obtained spectrogram is compared with a spectral library to determine a chemical structure and verified through a reference substance; the molecules and ions of the identified matters and myricetin, dihydromyricetin, catechinic acid / epicatechin, daidzein, glycitein and glycyrrhizin / isoliquiritigenin are adopted as characteristic ions for scanning, the molecules and ions of the searched glycosylation flavonoid matters are used as the parent ions for the second level mass spectrum scanning, an obtained spectrogram is compared with the spectral library to determine a chemical structure and verified through the reference substance. According to the invention, the method is suitable for the analyzing and detecting micro even trace flavonoid matters.

Description

technical field [0001] The invention belongs to the technical field of separation and identification of plant extracts, and in particular relates to a method for separation and identification of flavonoids in tobacco by liquid chromatography-mass spectrometry. Background technique [0002] Flavonoids, also known as flavonoids, refer to compounds whose basic core is 2-phenylchromone, and generally refer to a series of compounds in which two benzene rings with phenolic hydroxyl groups are connected to each other through the central three carbon atoms. Flavonoids are important secondary metabolites of plants, which have important effects on plants themselves and humans. In terms of plant stress resistance, flavonoids have shown significant effects in plant resistance to insects, viruses, and bacteria. In terms of the impact on human health, a large number of reports have shown that flavonoids have anti-cancer, anti-aging and anti-virus effects. Therefore, the isolation and id...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
Inventor 逄涛李勇卢秀萍师君丽陈萍
Owner YUNNAN ACAD OF TOBACCO AGRI SCI
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