RU2013125221A

RU2013125221A - HIGHLY SENSITIVE METHOD FOR DETERMINING THE NUMBER OF COMPONENTS OBTAINED FROM MEDICINAL HERBS

Info

Publication number: RU2013125221A
Application number: RU2013125221/15A
Authority: RU
Inventors: Каёко СУДЗУКИ; Цунехару СУДЗУКИ; Митико ЦУКАХАРА; Юко ХАРУТА; Акира ХАТТА; Юдзи ХАМАДА; Хидэо ИНОЭ
Original assignee: Минофаген Фармасьютикал Ко., Лтд.
Priority date: 2010-11-16
Filing date: 2011-11-15
Publication date: 2014-12-10
Also published as: JP2012107954A; KR20130076892A; KR101476144B1; WO2012067090A1; CN103210308B; CN103210308A; JP5193266B2; RU2558042C2

Abstract

1. Высокочувствительный способ определения количества компонентов, полученных из лекарственных трав, включающий:стадию экстракции, на которой экстракт, содержащий компоненты, полученные из лекарственных трав, готовят путем: введения смеси биологического образца с щелочью или спиртом в твердую фазу, обладающую обращено-фазовой распределительной функцией и функцией анионного обмена; промывания твердой фазы по крайней мере один раз очищающей жидкостью; и последующего элюирования из твердой фазы кислым спиртом, истадию количественного определения, на которой по меньшей мере один компонент, выбранный из группы, состоящей из глицирризина, глицирретиновой кислоты, метаболитов глицирризина и глицирретиновой кислоты, родственных соединений глицирризина и глицирретиновой кислоты, компонентов сапонина, содержащихся в лакричнике (солодке), и их фармакологически приемлемых солей, содержащийся в экстракте, полученном на стадии экстракции, детектируют и определяют количественно методом масс-спектрометрии;где очищающая жидкость представляет собой однокомпонентную жидкость или жидкую смесь по меньшей мере двух компонентов, выбранных из группы, включающей воду, щелочь, спирт и ацетонитрил, ина стадии экстракции твердую фазу промывают жидкой смесью щелочи, спирта и воды, жидкой смесью щелочи и воды или водой с последующим промыванием спиртом, жидкой смесью спирта и воды или ацетонитрилом.2. Высокочувствительный способ определения количества компонентов, полученных из лекарственных трав, по п. 1, в котором на стадии экстракции твердую фазу промывают жидкой смесью щелочи, спирта и воды с последующим промыванием спи�1. A highly sensitive method for determining the amount of components obtained from medicinal herbs, including: an extraction step in which an extract containing components obtained from medicinal herbs is prepared by: introducing a mixture of a biological sample with alkali or alcohol into a solid phase having a reversed-phase distribution function and function of anion exchange; washing the solid phase at least once with a cleaning liquid; and subsequent elution from the solid phase with acid alcohol, a quantitative determination, in which at least one component selected from the group consisting of glycyrrhizin, glycyrrhetinic acid, metabolites of glycyrrhizin and glycyrrhetinic acid, related compounds of glycyrrhizin and glycyrrhetinic acid, components of saponin contained in licorice (licorice), and their pharmacologically acceptable salts, contained in the extract obtained in the extraction stage, are detected and quantified by mass spectrometry; where the cleaning liquid is a one-component liquid or a liquid mixture of at least two components selected from the group consisting of water, alkali, alcohol and acetonitrile, at the extraction stage, the solid phase is washed with a liquid mixture of alkali, alcohol and water, a liquid mixture of alkali and water or water, followed by washing with alcohol, a liquid mixture of alcohol and water, or acetonitrile. 2. A highly sensitive method for determining the amount of components obtained from medicinal herbs according to claim 1, wherein in the extraction step the solid phase is washed with a liquid mixture of alkali, alcohol and water, followed by washing with alcohol

Claims

1. A highly sensitive method for determining the amount of components derived from medicinal herbs, including:

an extraction step in which an extract containing components obtained from medicinal herbs is prepared by: introducing a mixture of a biological sample with alkali or alcohol into a solid phase having a reversed-phase distribution function and anion exchange function; washing the solid phase at least once with a cleaning liquid; and subsequent elution from the solid phase with acid alcohol, and

a quantitative determination step in which at least one component selected from the group consisting of glycyrrhizin, glycyrrhetinic acid, metabolites of glycyrrhizin and glycyrrhetinic acid, related compounds of glycyrrhizin and glycyrrhetinic acid, components of saponin contained in licorice (licorice), and their pharmacologically acceptable the salts contained in the extract obtained in the extraction step are detected and quantified by mass spectrometry;

where the cleaning liquid is a single component liquid or a liquid mixture of at least two components selected from the group consisting of water, alkali, alcohol and acetonitrile, and

at the extraction stage, the solid phase is washed with a liquid mixture of alkali, alcohol and water, a liquid mixture of alkali and water or water, followed by washing with alcohol, a liquid mixture of alcohol and water or acetonitrile.

2. A highly sensitive method for determining the amount of components obtained from medicinal herbs according to claim 1, wherein in the extraction step the solid phase is washed with a liquid mixture of alkali, alcohol and water, followed by washing with alcohol, a liquid mixture of alcohol and water, or acetonitrile;

3. A highly sensitive method for determining the amount of components obtained from medicinal herbs according to claim 2, wherein in a liquid mixture of alkali, alcohol and water, 0.5-28 vol.% Aqueous ammonia and methanol are mixed in a ratio of from 99: 1 to 1 : 3.

4. A highly sensitive method for determining the amount of components derived from medicinal herbs, according to any one of paragraphs. 1-3, in which the biological sample is blood, blood plasma or tissue extract.