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Detection method for dammarane type four-ring triterpenoid saponin

A tetracyclic triterpene saponin and dammarane-type technology, which is applied in the detection field of dammarane-type tetracyclic triterpene saponins, can solve the problem that the mechanism of action in vivo cannot be evaluated and studied in depth, cannot be detected, and the concentration of saponins is low and other problems, to achieve the effect of excellent sensitivity and linear relationship

Inactive Publication Date: 2006-12-27
INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, due to the low concentration of saponins in plasma after oral administration of such saponins, the existing technology cannot detect them, so it is impossible to conduct in-depth evaluation and research on their bioavailability and in vivo mechanism of action.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1 Oral administration of commercially available notoginseng saponin tablets (Xuesaitong tablets) blood drug concentration detection test in beagle dogs

[0035] 1. Administration method: Beagle dogs were fasted for 12 hours before the test, and administered in the morning at a dose of 90 mg / kg of Panax notoginseng saponins. Take blank blood before taking the medicine, and take 3ml of foreleg venous blood at 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 16, 24, 36, and 48 hours after taking the medicine, anticoagulate with heparin, and centrifuge at 3000rpm immediately After 5 minutes, the plasma was separated and stored at -20°C for later use.

[0036] 2. Treatment of plasma samples: Accurately measure 0.5ml of plasma, centrifuge at 10,000r / min, absorb the supernatant and put it on a solid-phase extraction column, rinse with 2ml of double-distilled water, 2ml of 20% methanol, and wash with 2ml of chromatographic methanol The eluate was collected, dried under nitrogen at 60°C,...

Embodiment 2

[0111] Example 2 Detection test of blood drug concentration in Beagle dogs after oral administration of American ginseng capsules

[0112] Administration method: Beagle dogs were fasted for 12 hours before the test, and administered at a dose of 100 mg / kg American ginsenoside in the morning. Take blank blood before taking the medicine, and take 3ml of foreleg venous blood at 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 16, 24, 36, and 48 hours after taking the medicine, anticoagulate with heparin, and centrifuge at 3000rpm immediately After 5 minutes, the plasma was separated and stored at -20°C for later use.

[0113] Plasma sample processing: Accurately measure 0.5ml of plasma, put it in a centrifuge tube, add 2ml of methanol to precipitate protein, vortex and centrifuge at high speed, take supernatant and inject with a volume of 20μl, and record the chromatogram.

[0114] Analysis conditions:

[0115] HPLC conditions

[0116] Chromatographic column: SGE C18 column (50mm×2.1mm, 5μm)

...

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PUM

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Abstract

The invention relates to a method for detecting dammarane tetracyclic triterpenoid saponin and secondary aglycone. The inventive method uses high-efficiency liquid color spectrum-mass spectrum coupled device, wherein the conditions of high-efficiency liquid color spectrum comprises: color spectrum column is ODSC18 column; the column temperature is 20-38Deg. C; the conditions of mass spectrum comprise: the ion source is Turbo Ionspray source; the ejecting voltage is 3000-5000V; the Tem is 250-300Deg. C; the detector uses triplicate four-electrode serial mass spectrum detection and positive ion detection; the scanning method uses multi-reaction detection (MRM); CE is 17-36V; DP is 8-110V; the ion detection comprises: processing impact induce dissociation on the quasi-molecule ion peak of said saponin of first-stage scanning mass spectrum, to obtain the second-stage fragment ion; and uses multi-reaction detection (MRM) scanning method to analyze the ion reaction quantitatively, to represent the blood drug density of monomer saponin and secondary aglycone.

Description

technical field [0001] The invention provides a method for detecting samples containing a trace amount of dammarane-type tetracyclic triterpene saponins. technical background [0002] Modern studies have shown that the saponins in traditional Chinese medicines ginseng, American ginseng, and notoginseng are active ingredients, and they all contain dammarane-type tetracyclic triterpene saponins (Chinese medicine chemistry, Kuang Haixue P231-232), and their aglycones mainly contain 20(S) -Protopanaxadiol type and 20(S)-protopanaxatriol type, there are β-OH in the C3, C12 and C20 positions. The difference between protopanaxadiol type and protopanaxatriol type is only that the latter also has α-OH at the C6 position, and the positions of forming glycosides are usually at the C3, C20 and C6 positions. [0003] So far, 28 monomeric saponins have been isolated from different parts of Panax notoginseng F.H.Chen and himalaicus. Among them, there are 13 kinds of protopanaxatriol-type...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88G01N30/60G01N30/72G01N30/06
Inventor 朱春燕陈卫
Owner INST OF MEDICINAL PLANT DEV CHINESE ACADEMY OF MEDICAL SCI
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