Detection method for hydroxysafflor yellow A metabolic product

A technology of hydroxysafflower and detection method, which is applied in measuring devices, instruments, scientific instruments, etc., can solve the problem of in-depth research without metabolites and pathways, and achieve the effect of simplifying data processing

Active Publication Date: 2017-02-01
ZHEJIANG CHINESE MEDICAL UNIVERSITY
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  • Abstract
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Problems solved by technology

[0006] At present, the in vivo research on Guhong injection and safflower mainly selects the dynamic changes of the drug concentration in the body of the representative component hydroxysafflower yellow A (HSYA), draws the concentration curve of the drug, and carries out drug metabolism research in vivo, but all There is no in-depth research on the metabolites and pathways of drugs after they enter the body

Method used

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  • Detection method for hydroxysafflor yellow A metabolic product
  • Detection method for hydroxysafflor yellow A metabolic product
  • Detection method for hydroxysafflor yellow A metabolic product

Examples

Experimental program
Comparison scheme
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Embodiment 1

[0079] 1. Instrument conditions: liquid mass spectrometry LC-MS

[0080] Configuration: 1. Mass spectrometer: AB 4500

[0081] 2. Liquid chromatograph: Shimadzu LC-20ADXR

[0082] 2. Sample information

[0083] "Guhong injection" was intravenously injected into SD rats (about 200g / rat), and the injection volume was 12mg / kg. At 5min, 15min, 30min, 45min, 1h, 1.5h, 2h, 4h, 6h, and 8h, 0.5mL of orbital blood was collected, and the blood samples were placed in heparinized centrifuge tubes, and the temperature was 5000r·min at 4°C. -1 After centrifugation for 10 min, the plasma was separated and stored in a refrigerator at -80°C until analysis.

[0084] Blood sample processing method: draw 0.3mL of plasma into a 1.5mL centrifuge tube, accurately add 0.9mL of acetonitrile, vortex and mix for 2min, centrifuge at 6000r / min for 10min, take 0.6mL of supernatant, blow dry with nitrogen, 0.3 Dissolve in mL acetonitrile, centrifuge at 10000r / min, and take the supernatant for testing....

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Abstract

The invention discloses a detection method for hydroxysafflor yellow A metabolic products. The method includes the steps of: 1) performing intravenous injection to an SD rat with a safflower extract and aceglutamide injection, performing orbid blood sampling, and centrifugally separating the blood sample to obtain plasma; 2) sucking the plasma with a micro sample injector, adding acetonitrile with volume ratio of plasma to acetonitrile being 1:3-3.5, uniformly mixing the substances with vortex and centrifuging the mixture to obtain a supernatant, blow-drying the supernatant with nitrogen gas, dissolving the dried substance in acetonitrile, and centrifuging the solution to obtain a supernatant as a detection sample; and 3) detecting and analyzing the sample through high performance liquid chromatography-triple quadrupoles linear ion trap tandem mass spectrometry, and analyzing the metabolic products in a p-MRM-IDA-EPI mode. In the invention, a p-MRM method is employed in the mass spectrum with combination of Light Sight software to quickly identify and analyze the sample for tracing metabolic change of component change and structure change in the plasma; 11 hydroxysafflor yellow A metabolic products are found in the result. The method can be used for obtaining a time change curve of the hydroxysafflor yellow A and the metabolic product thereof in plasma, and successfully deduces and identifies the molecular structures of seven metabolic products and isomers of the hydroxysafflor yellow A. The method, compared with a chemical structure analytic method, is more quick and sensitive.

Description

technical field [0001] The invention belongs to the field of drug detection and analysis, and in particular relates to a detection method for detecting hydroxysafflower yellow A metabolite products in blood samples. [0002] Background technique: [0003] Safflower is the dried flower of Carthamustinctorius L., a plant in the Compositae family. Safflower yellow (SY) exists in the water-soluble extraction part of safflower, and is considered to be the main effect substance of safflower, among which the content of hydroxy safflower yellow A (HSYA) is relatively high, which is an effective monomer component that exerts pharmacological effects It can inhibit the platelet aggregation induced by adenosine diphosphate (ADP) or collagen, prevent the formation and development of thrombus or promote the dissolution of thrombus. [0004] Guhong injection (Guoyao Zhunzi H22026638) is a sterilized aqueous solution made of acetylglutamine and safflower extract. It is a commonly used clini...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
CPCG01N30/02
Inventor 万嘉洋张宇燕丁志山万海同黄平
Owner ZHEJIANG CHINESE MEDICAL UNIVERSITY
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