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Method for detecting phenolic acid substances in blood plasma

A technology for phenolic acids and plasma, applied in the field of detecting drug content in blood, can solve problems such as difficulty in separation, and achieve the effect of rapid method

Pending Publication Date: 2018-04-24
TIANJIN TASLY PHARMA CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Moreover, these compounds have similar properties and similar retention behaviors, making separation more difficult for these compounds

Method used

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  • Method for detecting phenolic acid substances in blood plasma
  • Method for detecting phenolic acid substances in blood plasma
  • Method for detecting phenolic acid substances in blood plasma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Embodiment 1: the establishment of analysis method

[0056] 1 Preparation of solution

[0057] 1.1 Series standard control solution preparation

[0058] Precisely weigh 10.00 mg of rosmarinic acid, Sal D, shikonic acid, and Sal B reference substances in a 5 mL volumetric flask, dissolve them in methanol and make up to a concentration of 2.00 mg·mL -1 The stock solutions of rosmarinic acid and Sal D were diluted with methanol to concentrations of 1, 2, 5, 20, 50, 200, 500, 1000, 5000ng·mL -1 A series of standard control solutions, shikonic acid and Sal B stock solutions were diluted with methanol to concentrations of 5, 10, 25, 100, 250, 1000, 2500, 5000, and 25000 ng·mL -1 A series of standard control solutions, stored at -20°C.

[0059] 1.2 Preparation of internal standard solution

[0060] Accurately weigh 0.5mg of chloramphenicol (IS) reference substance in a 10mL volumetric flask, dissolve it with methanol and make it to volume to make a concentration of 0.05mg·...

Embodiment 2

[0070] Confirmation of embodiment 2 analytical method

[0071] 1. Mass Spectrometry

[0072] Prepare certain concentrations of rosmarinic acid, Sal D, shikonic acid, Sal B, and chloramphenicol respectively. Under ESI ionization mode, rosmarinic acid, Sal D, shikonic acid, Sal B, and chloramphenicol mainly produce [M-H ]-Quasi-molecular ion peaks are m / z359.0, m / z 417.1, m / z537.0, m / z 717.0, m / z 321.1, and the main fragment ions are m / z 161.1, m / z175.0 , m / z 493.1, m / z 519.1, m / z 152.0, see the results figure 1 .

[0073] 2. Method specificity

[0074] Take 100 μL of blank plasma from 6 Wistar rats in a 1.5mL centrifuge tube, replace the internal standard with 10 μL of methanol, and perform LC-MS / MS analysis according to the law under the “Plasma Sample Processing” item. For details, see figure 2 , to obtain a chromatogram of a blank plasma sample as figure 2 A: Add a certain standard solution into blank plasma and internal standard solution, and operate in the same way ...

Embodiment 3

[0097] Embodiment 3: condition optimization

[0098] 1. Column selection

[0099] This test examines Waters ACQUITY UPLC HSS T3 (1.7μm, 2.1×100mm) T3, ACQUITY UPLC HSS C 18 (1.7μm, 2.1×100mm), CORTECS TM UPLC C 18(1.6μm, 2.1×100mm) 3 chromatographic columns. Waters ACQUITY UPLC HSS T3 and CORTECS TM UPLC C 18 For the separation effect of the four compounds, the column efficiency is higher. CORTECS TM UPLC C 18 The peak position of chloramphenicol on the inner side of the chromatographic column is relatively backward, but the sample residue is more serious when using the T3 chromatographic column, so CORTECSTM UPLC C 18 As the test column.

[0100] 2. Mobile phase selection

[0101] Compared methanol and acetonitrile, it was found that when acetonitrile was used as the organic phase, the chromatographic peak shape was better and the response was higher than that of methanol. The study also found that adding 0.1% formic acid to the water phase resulted in better chrom...

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Abstract

The invention relates to a method for detecting phenolic acid substances in blood plasma. The method includes steps of 1, preparing standard reference solution; 2, preparing internal standard solution; 3, preparing quality control (QC) samples; 4, treating medicated blood plasma samples; 5, carrying out detection. The method has the advantages that the plasma concentration of salvianolic acid B, salvianolic acid D, rosmarinic acid and lithospermic acid in blood plasma of rats is measured by the inventor by the aid of simple, convenient and sensitive LC-MS / MS (liquid chromatography-mass spectrometry / mass spectrometry) processes with high specificity after administration injection salvianolic acids are injected to the rats, and research is carried out on elimination relations of main effective components of different doses of the injection salvianolic acids in mammals.

Description

technical field [0001] The invention relates to a method for detecting drug content in blood, in particular to a method for detecting phenolic acid substances in blood plasma. Background technique [0002] The composition of traditional Chinese medicine is very complex. Even if it is a single medicine, it contains dozens of active ingredients. The active ingredients and mechanism of action of many traditional Chinese medicines are not very clear so far. In addition, the content of some active ingredients in Chinese medicines is very small, and there are many analogues with similar structures, which brings many difficulties to the study of pharmacokinetics. . Modern pharmacological research has preliminarily proved that the effect of multi-component drugs is not simply the addition of several single components or the subtraction of toxicity, but the synergy, antagonism or modification of the drugs in the prescription. To achieve the expected therapeutic purpose, it is obvio...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
CPCG01N30/02G01N30/06G01N2030/042
Inventor 佟玲李东翔谢秀满
Owner TIANJIN TASLY PHARMA CO LTD
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