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Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof

A detection method, the technology of Dian Chonglou, applied in the direction of measuring devices, instruments, scientific instruments, etc., can solve the problems of time spent on research and expensive analysis costs, and achieve low detection cost, high mass accuracy, and good mass axis stability Effect

Inactive Publication Date: 2015-07-08
CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chonglou saponin is the main medicinal ingredient of Chonglou. There are 4 kinds of Chonglou saponin components listed in the Chinese Pharmacopoeia, and there are 5 kinds of Chonglou saponin components found in literature reports. The identification method of saponins is mainly to use column chromatography to separate monomer saponins and collect various monomer saponins, and then use nuclear magnetic resonance spectrometer for structure identification, which takes more than several months of research time and expensive analysis cost

Method used

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  • Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof
  • Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof
  • Detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] a. Sample extraction: Accurately weigh 0.2 g of Scutellaria dioica sample, dissolve it in 5 ml of methanol, extract it ultrasonically for 30 minutes, make a sample solution containing 0.04 g / ml of Scutellaria dianthus saponin, and measure after centrifuging through a 0.2 μm filter membrane;

[0015] b. Liquid chromatography conditions:

[0016] Instrument: Ultra High Phase Liquid Chromatography

[0017] Chromatographic column: C18 column (150×2.1mm, 5μm)

[0018] Mobile phase: A is water phase: pure water, B is organic phase: acetonitrile

[0019] Gradient condition:

[0020]

[0021] Flow rate: 0.4mL / min, injection volume: 10μL, detection wavelength: 203nm, column temperature: 30°C

[0022] c. Mass Spectrometry Conditions

[0023] Instrument: Quadrupole-Electrostatic Field Orbitrap High Resolution Mass Spectrometry

[0024] Mass spectrometry parameters: ionization mode HESI (+), spray voltage +3.2KV / -3.0KV, sheath gas 40arb, auxiliary gas 10arb, capillary heati...

Embodiment 2

[0034] a. Sample extraction: Accurately weigh 0.2 g of Scutellaria dioica sample, dissolve it in 5 ml of methanol, extract it ultrasonically for 30 minutes, make a sample solution containing 0.04 g / ml of Scutellaria dianthus saponin, and measure after centrifuging through a 0.2 μm filter membrane;

[0035] b. Liquid chromatography conditions:

[0036] Instrument: Ultra High Phase Liquid Chromatography

[0037] Chromatographic column: C18 column (150×2.1mm, 5μm)

[0038] Mobile phase: A is water phase: pure water, B is organic phase: acetonitrile

[0039] Gradient condition:

[0040]

[0041] The flow rate is 0.1min, the detection wavelength is 190nm, the injection volume is 2μL, and the column temperature is controlled at 20°C;

[0042] c. Mass Spectrometry Conditions

[0043] Instrument: Quadrupole-Electrostatic Field Orbitrap High Resolution Mass Spectrometry

[0044] Mass spectrometry parameters: ionization mode HESI (+), spray voltage +3.2KV / -3.0KV, sheath gas 40ar...

Embodiment 3

[0047] a. Sample extraction: Accurately weigh 0.2 g of Scutellaria dioica sample, dissolve it in 5 ml of methanol, extract it ultrasonically for 30 minutes, make a sample solution containing 0.04 g / ml of Scutellaria dianthus saponin, and measure after centrifuging through a 0.2 μm filter membrane;

[0048] b. Liquid chromatography conditions:

[0049] Instrument: Ultra High Phase Liquid Chromatography

[0050] Chromatographic column: C18 column (150×2.1mm, 5μm)

[0051] Mobile phase: A is water phase: pure water, B is organic phase: acetonitrile

[0052] Gradient condition:

[0053]

[0054] The flow rate was 0.8min, the detection wavelength was 210nm, the injection volume was 20μL, and the column temperature was controlled at 45℃.

[0055] c. Mass Spectrometry Conditions

[0056] Instrument: Quadrupole-Electrostatic Field Orbitrap High Resolution Mass Spectrometry

[0057] Mass spectrometry parameters: ionization mode HESI (+), spray voltage +3.2KV / -3.0KV, sheath gas ...

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Abstract

The invention discloses a detection method used for rapid identification of paris polyphylla var. yunnanensis saponin composition, and applications thereof. The detection method, which is used for rapid identification of unknown paris polyphylla var. yunnanensis saponin compounds by liquid chromatography / high-resolution mass spectrometry, comprises following steps: quadrupole-electrostatic field orbitrap high resolution mass spectrum is used for detection; the pyrolysis laws of known paris polyphylla var. yunnanensis saponin compounds are analyzed; tandem mass spectrometry fragments are obtained by high-energy collisions induced dissociation (HCD); and more fragments with lower weight which are quite helpful for structural analysis of compounds are obtained by conventional collision induced dissociation (CID). It is found that paris polyphylla var. yunnanensis comprises 17 saponin compounds, wherein 3 new steroidal saponins are reported for the first time, and 5 saponin compounds are newly discovered compounds of paris polyphylla var. yunnanensis saponin.

Description

technical field [0001] The invention relates to a detection method of traditional Chinese medicine components, in particular to a detection method for rapidly identifying the composition of pachyphyllum saponins and its application. Background technique [0002] Dianzhonglou is a rare variety of medicinal materials in Yunnan, and it is one of the two basic source varieties of Zhonglou medicinal materials in "Chinese Pharmacopoeia" (2010 edition one). It is the main raw material of more than 50 kinds of famous and high-quality Chinese patent medicine products in my country. Chonglou saponin is the main medicinal ingredient of Chonglou. There are 4 kinds of Chonglou saponin components listed in the Chinese Pharmacopoeia, and there are 5 kinds of Chonglou saponin components found in literature reports. The identification method of saponins is mainly to use column chromatography to separate monomer saponins and collect various monomer saponins and then use nuclear magnetic reson...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/88
Inventor 彭迎霞马晓刚
Owner CHECKOUT & QUARANTINE TECH CENT YUNNAN ENTRY &EXIT CHECKOUT & QUARANTINE BUR