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Qualitative and quantitative method of triterpene saponins in traditional Chinese medicine by electrospray protonation fragmentation-mass spectrometry multiple reaction detection mode

A technology for monitoring triterpenoid saponins and multiple reactions, which is applied in the field of chemical analysis and detection, and can solve the problems of low detection sensitivity and difficulty in quantification of traditional Chinese medicine saponins.

Active Publication Date: 2020-04-10
匡海学 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

While using electrospray tandem mass spectrometry in MRM mode, fragmentation of [M+Na] + and [M-H] - Precursor ions can achieve satisfactory quantitative results for the constant triterpene saponins in traditional Chinese medicine. However, the detection sensitivity of this method is low when detecting some traditional Chinese medicine saponins, and the quantification is difficult.

Method used

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  • Qualitative and quantitative method of triterpene saponins in traditional Chinese medicine by electrospray protonation fragmentation-mass spectrometry multiple reaction detection mode
  • Qualitative and quantitative method of triterpene saponins in traditional Chinese medicine by electrospray protonation fragmentation-mass spectrometry multiple reaction detection mode
  • Qualitative and quantitative method of triterpene saponins in traditional Chinese medicine by electrospray protonation fragmentation-mass spectrometry multiple reaction detection mode

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1 Establishment of a Quantitative Method for Triterpenoid Saponins from Paeoniae Ruba

[0038] 1 experimental equipment

[0039] Qtrap-4000 triple quadrupole linear ion trap mass spectrometer (AB SCIEX company, the United States); H-class ultra-high performance liquid chromatography (online degasser, automatic sampler, quaternary pump, American Waters company); ACQUITY UPLC HSS T3 chromatographic column (2.1×150mm, 1.8μm), connected to a Waters HSS T3 guard column (2.1×5mm, 1.8μm). Milli-Q water purifier (Millipore China Co., Ltd.), ML104 / 02 electronic analytical balance (Mettler Toledo Instruments Shanghai Co., Ltd.), Vortex 3000 vortex oscillator (Wiggens, Germany); Mikro 200R centrifugal Machine (Hettich, Germany), KQ-500DB CNC ultrasonic cleaner (Kunshan Ultrasonic Instrument Co., Ltd.).

[0040] 2 Preparation of standard products

[0041] Accurately weigh 1-9 kinds of saponin standard items ( figure 1 Shown) appropriate amount, accurately weighed, add m...

Embodiment 2

[0063] Example 2 Establishment of Liaodong Aralia triterpene saponin quantitative method

[0064] 1. Experimental equipment

[0065] With embodiment 1.

[0066] 2. Preparation of standard products

[0067] Accurately weigh 1-10 saponin standard items ( image 3 Shown) appropriate amount, accurately weighed, add methanol and be mixed with the mixed reference solution that mass concentration is respectively 1.69, 0.21, 0.20, 0.28, 1.00, 0.98, 1.04, 1.00, 1.00, 1.00mg / ml as stock solution, in 4 Store at ℃ for later use.

[0068] 3. Optimizing DP, CE and CXP mass spectrometry parameters based on response surface center composition design

[0069] 3.1 Overall response surface Box-Behnken experimental design

[0070] Using mass spectrometry in multiple reaction monitoring (MRM) mode, using saponins 1-10 as template compounds, the response value and reproducibility of proton / ammonium ion cleavage targeted ion pairs were optimized by response surface methodology. According to th...

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Abstract

The invention discloses a method for performing qualitation and quantification on triterpenoid saponin in traditional Chinese medicine by utilizing an electrospray protonation pyrolysis-mass spectrummulti-reaction detection mode. The triterpenoid saponin can form a stable [M+H]<+> and / or [M+NH4)<+> molecular ion peak in a flow phase containing formic acid or ammonium formate. According to the method disclosed by the invention, a liquid chromatogram-tandem quadrupole mass spectrometry combined instrument is utilized, an electrospray positive ion tandem mass spectrum detection mode is utilized,and mass spectrum parameters of declustering potential, impact energy, impact chamber injection voltage and the like can be quickly optimized in an online mode by a single factor and response surfacemethod; thus, high-strength triterpenoid saponin protonation pyrolysis MRM target ion pairs can be formed; a Q1 calculation formula is [M+H]<+> or [M+NH4]<+>, and a Q3 calculation formula is [aglycone +H-nH2O]<+>. By means of the series of characteristic MRM target ion pairs, the purpose of quick qualitative and quantitative detection on the triterpenoid saponin in the traditional Chinese medicine is realized. The method disclosed by the invention provides an effective technology method for the qualitative and quantitative detection on the triterpenoid saponin in the traditional Chinese medicine and has been successfully applied to content measurement on the triterpenoid saponin of caulophyllum robustum and aralia elata.

Description

technical field [0001] The invention relates to a method for quantifying triterpene saponins in traditional Chinese medicine, in particular to a method for qualitatively and quantitatively determining triterpene saponins in traditional Chinese medicine based on electrospray protonation cracking-mass spectrometry multiple reaction detection mode. The invention belongs to the technical field of chemical analysis and detection. Background technique [0002] Triterpene saponins have been widely accepted by researchers as the main effective parts or groups of effective parts in traditional Chinese medicine. However, triterpene saponin molecules lack chromophores, absorb ultraviolet light at the end, and are non-volatile. Therefore, there is currently a lack of effective methods for the quantification of triterpene saponins. At present, the quantitative methods of triterpene saponins in traditional Chinese medicine mainly use high performance liquid chromatography-ultraviolet spe...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 夏永刚梁军匡海学杨炳友
Owner 匡海学
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