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Quantitative analysis method for plasma concentration of musk ketone in artificial musk

A technology of artificial musk and blood drug concentration, applied in the direction of analyzing materials, measuring devices, material separation, etc., can solve the problems of analysis result error, loss, inconvenience of large sample detection, etc.

Inactive Publication Date: 2013-10-23
MACAU UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The analytical method for determining muscone in rats after intragastric administration of artificial musk has not been reported before. It mainly involves direct intragastric administration and intravenous injection of muskone. The analytical method is mainly gas chromatography, and the internal standard used is different tonic For osteolipin and n-nonadecane, the biological sample processing method is to extract three times with ether and combine the extracts and N 2 Blow dry, add volatile solvent trichloromethane or n-hexane to measure
The problem that this analysis method exists is that the biological sample processing step is more loaded down with trivial details, and muscone is a volatile substance, firstly there is volatilization loss in the process of extracting three times with ether, secondly with N 2 There is also loss in the process of drying and reconstitution, resulting in errors in analysis results
Such complex processing steps are also inconvenient for large-sample detection in pharmacokinetic studies
[0007] Muscone is rapidly metabolized in the human body after being absorbed by the gastrointestinal tract, and the plasma concentration of its prototype drug is extremely low, with a Cmax of about 3.14mg.L -1 , while the content of muscone in artificial musk is only about 0.86%, which is more difficult to measure

Method used

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  • Quantitative analysis method for plasma concentration of musk ketone in artificial musk
  • Quantitative analysis method for plasma concentration of musk ketone in artificial musk
  • Quantitative analysis method for plasma concentration of musk ketone in artificial musk

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Solution preparation

[0024] Reference substance series solution

[0025] Accurately weigh 0.0567g of the muscone standard substance, put it in a 100ml volumetric flask and dilute to the mark with methanol, and use it as the muscone standard substance stock solution (567μg / ml). Dilute the stock solution with methanol successively to prepare muskone standard solutions with concentrations of 0.756, 1.134, 5.67, 11.34, 56.7, and 113.4 μg / ml. The solution was placed in a refrigerator at 4°C and stored for later use.

[0026] Internal standard working solution

[0027] Accurately weigh 0.0117 g of cyclohexanone, prepare an internal standard stock solution with a concentration of 117 μg / ml with methanol, and dilute the stock solution with methanol to obtain an internal standard solution with a concentration of 53.7 μg / ml. The solution was placed in a refrigerator at 4°C and stored for later use.

[0028] (2) Preparation of quality control samples

[0029] Take 0.3ml...

Embodiment 2

[0038] (1) Solution preparation

[0039] The preparation of reference substance series solutions and quality control samples is the same as in Example 1.

[0040] (2) Plasma sample processing

[0041] After intragastric administration of artificial musk to rats within a certain period of time, blood was taken from the eye socket and placed in a heparinized test tube. After centrifuging the obtained blood, the upper plasma sample was obtained. Take 0.3ml of the plasma sample, add 0.3ml of acetonitrile, and add the internal standard solution at the same time (Alcoholic solution of cyclohexanone at 53.7 μg / ml) 20 μl, mix and vortex for 100 s, then centrifuge at 8000 r / min for 5 min, take the supernatant and put it in a 10 ml headspace sampling vial, then add 2 ml of 3wt% NaCl solution to seal and then test .

[0042] (3) Measurement conditions

[0043] Agilent 6890N gas chromatograph was used, connected with Agilent 5975B mass spectrometer, and Agilent G1888A headspace sampler...

Embodiment 3

[0048] (1) Solution preparation

[0049] The preparation of reference substance series solutions and quality control samples is the same as in Example 1.

[0050] (2) Plasma sample processing

[0051] After intragastric administration of artificial musk to rats within a certain period of time, blood was taken from the eye sockets and placed in a heparinized test tube. The obtained blood was centrifuged to obtain the upper layer of plasma samples. 0.3ml of plasma samples were taken, 0.3ml of acetonitrile was added, and an internal standard ring was added at the same time. Mix 20 μl of hexanone alcohol solution, mix and vortex for 120 s, centrifuge at 6000 r / min for 7 min, take the supernatant and put it in a 10 ml headspace sampling vial, add 2 ml of 3wt% NaCl solution and seal it for testing.

[0052] (3) Measurement conditions

[0053] Agilent 6890N gas chromatograph was used, connected with Agilent 5975B mass spectrometer, and Agilent G1888A headspace sampler was used toge...

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Abstract

The invention provides a quantitative analysis method for plasma concentration of musk ketone in artificial musk. The quantitative analysis method comprises the following steps of: adding acetonitrile and an internal standard working solution into plasma of a rat lavaged by the artificial musk, mixing, vortexing, centrifuging, taking supernatant and filling in a headspace sampling bottle of a headspace sampler to test and also adding 1-5 percent by weight of NaCl aqueous solution into the headspace sampling bottle, wherein the internal standard solution is alcoholic solution of cyclohexanone or normal hexane; and determining by using a gas chromatography-mass spectrometry method. According to the quantitative analysis method, the headspace sampling gas chromatography-mass spectrometry method is adopted, the method for precipitating protein by using the acetonitrile is used as a biological sample pretreatment method, and the determination is performed under proper temperature programming conditions and at proper headspace balance temperature and proper headspace balance time. The determination method has high specificity and high sensitivity, is quick and convenient to determine, can greatly simplify an experimental process and is suitable for pharmacokinetic study.

Description

technical field [0001] The present invention relates to the analysis method of biological substance, more specifically relates to the blood drug concentration quantitative analysis method of muscone (muscone) in artificial musk. Background technique [0002] Facing the increasingly severe situation of musk deer resources, SFDA determined and announced the list of Chinese patent medicines that need to use natural musk according to the status of natural musk and bear bile powder resources and the quantity of raw materials in stock notified by the State Forestry Administration. And those who have not been approved will no longer be able to use natural musk. The country's strict management of natural musk has provided market opportunities for artificial musk that was recognized by the Ministry of Health as "used in the same formula as natural musk" as early as 1994. Artificial musk is a new class of traditional Chinese medicine successfully developed in my country in 1993. It h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/14G01N30/16G01N30/30G01N30/34G01N30/32G01N30/88
Inventor 吴其标李海涛
Owner MACAU UNIV OF SCI & TECH
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