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Method for separating and purifying in-vivo metabolites of protopanaxadiol

A technique for protopanaxadiol and metabolites is applied in the field of separating and purifying metabolites of protopanaxadiol in fecal samples, and achieves the effects of easy collection, low price, easy collection and accumulation

Inactive Publication Date: 2017-07-04
FUDAN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] In the prior art, since ginsenosides have a certain cracking law in the mass spectrometer detector, and their metabolites have similar structures and have similar cracking rules, the use of mass spectrometry data to infer the structure of metabolites has certain limitations and limitations. Uncertainty, based on this, the inventor of the present application intends to establish a method for separating and purifying metabolites of protopanaxadiol in vivo, so as to further prepare pure metabolites, and then provide a basis for NMR detection to determine the structure of unknown compounds, as well as for Protopanaxadiol metabolism in vivo provides a reliable means of analysis

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  • Method for separating and purifying in-vivo metabolites of protopanaxadiol

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Embodiment 1

[0017] 1. Dosing regimen

[0018] 1.1 PPD suspension preparation

[0019] Add 0.5 g of sodium carboxymethyl cellulose to 100 mL of deionized water, heat and stir in a water bath at 40°C until it is completely dissolved. Take 500 mg of PPD raw material, add 100 mL of the above 0.5% sodium carboxymethyl cellulose solution, mix thoroughly and store at 4°C for later use;

[0020] 1.2 Administration and sample collection

[0021] Take 10 male S.D. rats, each with a body weight of 250-300 g, and place them in metabolic cages. After adaptive feeding for 1 week, 50 mg / kg of body weight is administered orally once a day at 9 o'clock in the morning. Medicine for 40 days; from day 2, collect the feces in the metabolic cage every day. After the rat feces are collected, try to pick out the food residues, and then put them in an oven to bake at a low temperature of 40°C, and dry them and store them in a sealed seal at -20°C;

[0022] 1.3 Crude extraction of metabolites

[0023] Take 2kg...

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Abstract

The invention relates to a method for separating and purifying in-vivo metabolites of protopanaxadiol in excrement, belonging to the field of pharmacoanalysis. The method employs a silica-gel chromatographic column for crude separation and then employs mass spectrometry-based preparative liquid chromatography for purification; and through combination of the above two separation methods, pure metabolites are prepared. According to the method, separation via the silica-gel chromatographic column and separation via preparative liquid chromatography are successively applied so as to rapidly and efficiently purify a plurality of pure metabolites in one shot; gradient elution conditions of the silica-gel chromatographic column and the preparative liquid chromatography enable a plurality of unknown substances to be separated and purified in a short period of time; the method uses excrement us a to-be-separated sample, and the to-be-separated sample is low in price, easy to acquire and accumulate and convenient for separation and purification of a great number of pure metabolites; and a foundation is further provided for nuclear magnetic resonance detection and determination of the structures of unknown compounds, and a reliable analysis means is provided for the process of in-vivo metabolism of protopanaxadiol.

Description

technical field [0001] The invention belongs to the field of pharmaceutical analysis, relates to a separation and purification method, in particular to a method for separation and purification of metabolites of protopanaxadiol in feces samples. Background technique [0002] The prior art discloses that protopanaxadiol is an in vitro hydrolyzate of protopanaxadiol type ginsenosides, and structurally, it belongs to dammarane type tetracyclic triterpenoid ginsenoside compounds, and its molecular formula is C 30 h 52 o 3 , with a molecular weight of 460.70 and a melting point of 197.5-198.5 degrees Celsius. Studies have shown that it has pharmacological activities similar to ginsenosides, including anti-tumor effects, anti-epileptic seizures, and anti-depressant effects. At present, protopanaxadiol has entered the stage of clinical trials as a new antidepressant drug. Therefore, the study of its metabolism in the human body plays an important role in ensuring the safety and re...

Claims

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Application Information

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IPC IPC(8): G01N1/34G01N30/14
CPCG01N1/34G01N30/14
Inventor 段更利郁颖佳凌今龙佳坤
Owner FUDAN UNIV
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