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Method for determining hydroxytyrosol in Beagle dog plasma

A technology of hydroxytyrosol and plasma, applied in the field of determination of hydroxytyrosol in Beagle dog plasma, can solve the problems of poor stability

Inactive Publication Date: 2021-01-08
SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

And for the content assay method of hydroxytyrosol in Beagle dog blood plasma, do not see bibliographical information yet, because hydroxytyrosol is very easily oxidized, and stability is poor, the biological analysis method that pharmacokinetic research requires must be sensitive and accurate, and the present invention establishes and utilizes for the first time LC / MS / MS method for the determination of hydroxytyrosol in Beagle dog plasma, laying a solid bioanalytical foundation for the preclinical study of hydroxytyrosol

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  • Method for determining hydroxytyrosol in Beagle dog plasma
  • Method for determining hydroxytyrosol in Beagle dog plasma
  • Method for determining hydroxytyrosol in Beagle dog plasma

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Experimental program
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Embodiment 1

[0022] The methodological investigation of embodiment 1 detection method of the present invention

[0023] 1. Preparation of standard solution

[0024] (1) Preparation of standard series solutions

[0025] Accurately weigh an appropriate amount of hydroxytyrosol, add solvent (containing 0.1% formic acid, 5mM ammonium acetate, 5mM ascorbic acid aqueous solution) and quantitatively dilute to prepare a 4mg / mL standard stock solution. Precisely measure an appropriate amount of standard stock solution, dilute it with a solvent gradient, and prepare a series of standard solutions with concentrations of 10, 20, 50, 200, 500, 2000, 5000, 10000, and 20000 ng / mL.

[0026] (2) Preparation of internal standard solution

[0027] Accurately weigh an appropriate amount of tolbutamide, add acetonitrile to dissolve and quantitatively dilute to prepare a 10 mg / mL tolbutamide stock solution. Precisely measure an appropriate amount of stock solution, and gradually dilute it with acetonitrile t...

Embodiment 2

[0052] Embodiment 2 Hydroxytyrosol pharmacokinetic experiment in Beagle dog body

[0053] The experiment adopted a two-period crossover design. 4 Beagle dogs, male, were randomly divided into 2 groups. After fasting for 12 hours, the intravenous injection group was given hydroxytyrosol solution by intravenous injection, and the oral group was given hydroxytyrosol capsules orally at a dosage of 14 mg / kg. About 1.5 mL of blood was collected from the canine forelimb vein in each group at 0.083, 0.167, 0.333, 0.5, 1, 2, 4, 6, 8 and 24 hours after administration.

[0054] The blood samples were placed on ice after collection, and the plasma was centrifuged within 1 hour (centrifugation conditions: 8000 rpm, 10 min, 2-8°C). After 14 days, the animals of the two groups were cross-administered, and the venous blood was collected and the plasma was prepared according to the above method. The collected plasma was stored in a -80°C refrigerator before analysis.

[0055] The determina...

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Abstract

The invention relates to a method for determining hydroxytyrosol in Beagle dog plasma, and belongs to the field of biological detection. According to the method, the concentration of hydroxytyrosol ina Beagle dog plasma sample is detected by adopting high performance liquid chromatography-electrospray ionization tandem mass spectrometry after butylurea tosylate is adopted as an internal standardand acetonitrile is adopted for protein precipitation treatment. The accuracy, precision, specificity, stability, extraction recovery rate, matrix effect and the like of the method all meet the requirements of Chemical Drug Non-Clinical Pharmacokinetic Research Technical Guidance principles issued in 2015 edition of Chinese Pharmacopoeia and NMPA2014 on biological sample analysis.

Description

technical field [0001] The invention relates to a method for measuring hydroxytyrosol in Beagle dog plasma, belonging to the field of biological detection. Background technique [0002] Hydroxytyrosol (HT), whose chemical name is 3,4-dihydroxyphenylethanol, is a natural polyphenolic component present in olive leaves and olive oil, has in vitro antioxidant effects, and is often used in health products and food additives . Recent studies have shown that hydroxytyrosol has various pharmacological activities beneficial to human health, such as anticancer, antibacterial, anti-inflammatory, antioxidative, and heart protection. Its medicinal value needs to be developed urgently. It will be the main research direction in the future. The preclinical research on the druggability evaluation of hydroxytyrosol involves pharmacokinetic / toxicokinetic studies of different species of experimental animals, so it is very important to establish a rapid, sensitive and specific hydroxytyrosol a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/88
CPCG01N30/88G01N2030/884
Inventor 房绍英孙淑萌李帅广程艳玲郑晓霞黄艳妮赵丹刘纯吕慧敏何小华马玉奎刘飞凌沛学
Owner SHANDONG ACADEMY OF PHARMACEUTICAL SCIENCES
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