Method for measuring absorption and transporting quantities of effective ingredients in herba-erigerontis and red-peony-root extract in Caco-2 cell model

A cell model and active ingredient technology, applied in measurement devices, material separation, material analysis, etc., can solve the problems of few reports on the absorption mechanism and unclear absorption process.

Inactive Publication Date: 2016-11-23
GUIZHOU MEDICAL UNIV
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Problems solved by technology

Although those skilled in the art have done a lot of research on the chemical components, pharmacological effects, and in vivo pharmacokinetic characteristics of

Method used

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  • Method for measuring absorption and transporting quantities of effective ingredients in herba-erigerontis and red-peony-root extract in Caco-2 cell model
  • Method for measuring absorption and transporting quantities of effective ingredients in herba-erigerontis and red-peony-root extract in Caco-2 cell model
  • Method for measuring absorption and transporting quantities of effective ingredients in herba-erigerontis and red-peony-root extract in Caco-2 cell model

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Embodiment 1

[0036] 1 Instruments and materials

[0037] 1 material

[0038] Xinshao extract (self-made, batch number 20150615); puerarin (batch number 0752-9605, purity>98%), gallic acid (batch number 110831-201204, purity>98%), caffeic acid (batch number 110885-200102, purity>98%) %), scutellarin (batch number 110842-201207, purity>98%), scutellarin (batch number 1384-101215, purity>98%) were purchased from China National Institutes for Food and Drug Control, paeonifloride (batch number S37-100126 , purity>98%) was purchased from the National Engineering Research Center of Traditional Chinese Medicine Solid Preparation Manufacturing Technology; Caco-2 cell line (ATCC); penicillin-streptomycin, 0.25% trypsin-0.02% EDTA digestion solution, Hank's buffer solution, DMEM medium, non-essential amino acids (Gibco); Coomassie brilliant blue (Nanjing Jiancheng Bioengineering Institute); fetal bovine serum (Hangzhou Sijiqing Bioengineering Materials Co., Ltd.); MTT, Triton X-100, DMSO (Beijing Ro...

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Abstract

The invention discloses a method for measuring the absorption and transporting quantities of effective ingredients in herba-erigerontis and red-peony-root extract in a Caco-2 cell model. the method includes the steps that herba-erigerontis and red-peony-root extract testing liquid is prepared; a series of standard solutions serving as reference samples are prepared from albiflorin std, gallic acid, caffeic acid, scutellarin, scutellarin and apigenin-7-o-glucronide; an internal standard solution is prepared from puerarin; then the human-source intestinal-gland cancer cell line Caco-2 cell model is built; the obtained testing liquid and the series of standard solutions are respectively added into the Caco-2 cell model, and after a cell lysis solution is added for cell lysis, a medicine-containing cell suspension is obtained; the absorption quantities of five active ingredients in the obtained medicine-containing cell suspension are measured through a UPLC-MS. According to the method, in the test, the Caco-2 cell model is combined with the LC-MS technology to research the absorption mechanism of the five active ingredients in the herba-erigerontis and red-peony-root extract, the characteristics of taking, transmembrane transporting and the like of the ingredients are studied, and the comprehensive absorption information of medicine molecules is provided in the cell level.

Description

technical field [0001] The invention belongs to the technical field of detection of traditional Chinese medicines, and in particular relates to a method for measuring the absorption and transport amount of active ingredients in Xinshao extract in a Caco-2 cell model. Background technique [0002] Xinshao Group Prescription, derived from Guizhou folk prescriptions, is composed of Erigeron breviscapus and Radix Paeoniae Rubra. It is decocted and taken in the folks for the treatment of hemiplegia, and has a good clinical basis. According to the etiology and pathogenesis of wind-phlegm flowing through the meridians, blood stasis blocked, meridian tunnels blocked, qi unable to flow, and blood unable to moisten, so limbs are useless instead of hemiplegia, the method of promoting blood circulation, removing stasis and dredging collaterals is adopted. Taking Erigeron breviscapus as the ruler, dissipating stasis and dredging collaterals, Radix Paeoniae Rubra, cooling blood heat, dred...

Claims

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Application Information

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IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/027G01N2030/8809
Inventor 王爱民兰燕宇黄勇巩仔鹏李月婷郑林刘亭董莉陈思颖
Owner GUIZHOU MEDICAL UNIV
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