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Method for detecting nine alkaloids in blood by liquid chromatography-tandem mass spectrometry and application

A technology of tandem mass spectrometry and liquid chromatography, applied in the field of liquid chromatography tandem mass spectrometry detection of 9 kinds of alkaloids in blood, can solve the problems of superposition of toxicity and low toxic dose, and achieves reduction of analysis cost, good economic benefits, and saving detection. cost effect

Active Publication Date: 2022-07-22
BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] From the above analysis, it can be seen that the above-mentioned various alkaloids not only have low toxic doses, but also may have superimposed toxicity in the human body. Therefore, it is necessary to detect the blood drug concentration of the above-mentioned alkaloids in the human body.

Method used

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  • Method for detecting nine alkaloids in blood by liquid chromatography-tandem mass spectrometry and application
  • Method for detecting nine alkaloids in blood by liquid chromatography-tandem mass spectrometry and application
  • Method for detecting nine alkaloids in blood by liquid chromatography-tandem mass spectrometry and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0124] (1) Preparation of standard solution and calibration of standard curve.

[0125] 1. Preparation of standard stock solution

[0126] Precisely weigh 9 kinds of alkaloid reference substances (not less than 1.00 mg), and after correction by the correction coefficient, add the standard dilution solution with a pipette to prepare a stock solution of 1 mg / mL, and store it in a -20 ℃ refrigerator.

[0127] 2. Preparation of reference substance mixed stock solution

[0128] Pipette 100 μL of 9 kinds of alkaloid stock solutions (1 mg / mL) each, pipette 100 μL of standard dilution, prepare 100 μg / mL reference substance mixed stock solution and store in -20 ℃ refrigerator .

[0129] 3. Preparation of standard working solution

[0130] Pipette an appropriate amount of the reference substance mixed stock solution (100 μg / mL), dilute it with the standard substance diluent step by step, prepare the standard curve working solution of each concentration, and store it in a -20 ℃ refrig...

experiment example 1

[0159] This experimental example is used to illustrate the personnel differences of the detection method according to the embodiment of the present invention:

[0160] Using the method of Example 1, 2 experimenters, each experimenter pretreated 6 samples with three concentrations of low, medium and high, respectively, to investigate the differences among the personnel, and the experimental results obtained are shown in Table 11.

[0161] Table 11: Differences among persons (P=2, n=6)

[0162]

[0163] According to Table 11, the CV is between 2.79% and 7.13%. The average accuracy is between 98.08% and 112.07%, indicating that there is little difference between personnel.

experiment example 2

[0165] (1) Linear range and lower limit of quantification:

[0166] The mixed standard solutions of 9 concentrations were prepared respectively and the chromatograms were obtained according to the above method. The standard curve was obtained by plotting the chromatographic peak area-concentration. The results showed that the linear ranges of the 9 alkaloids were shown in Table 12, and the lower limits of quantification (LLOQ) of the 9 alkaloids in the plasma samples were shown in Table 12.

[0167] Table 12: Intraassay Accuracy and Precision of Quantitative Lower Limits (n=6)

[0168]

[0169] According to Table 12, the 9 alkaloids in the plasma samples were in the range of 1 ng / mL to 200 ng / mL, with good linearity, and the correlation coefficient was R > 0.9900. The lower limit of quantification (LLOQ) for the nine alkaloids in plasma samples was 1 ng / mL. The lower limit of quantification (LLOQ) of the intra-assay precision test result CV<20%, which meets the test requi...

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Abstract

The invention relates to the technical field of medicine detection, in particular to a method for detecting nine alkaloids in blood through liquid chromatography-tandem mass spectrometry and application. The nine alkaloids comprise sophocarpidine, oxymatrine, cytisine, N-methyl cytisine, alpha-solanine, alpha-canthine, solasonine, solamargine and beta-solamargine; the method comprises the following steps: preparing a standard curve equation, processing to obtain a to-be-detected sample, and detecting the to-be-detected sample. The method can be used for simultaneously detecting nine alkaloids, has the characteristics of high accuracy and precision and wide detection range, can reduce the analysis cost, save manpower and analysis time, reduce the blood consumption and increase the compliance of subjects.

Description

technical field [0001] The invention relates to the technical field of medical detection, in particular to a method and application for detecting 9 kinds of alkaloids in blood by liquid chromatography tandem mass spectrometry. Background technique [0002] Alkaloids are alkaline nitrogen-containing organic compounds in living organisms, most of which exist in plants and some in animals. It has a cyclic structure, is insoluble in water, and can form salts with acids. It has certain optical activity and absorption spectrum. Most of them have a bitter taste and are colorless crystals, and a few are liquids. Many Chinese medicinal materials and many common toxic foods contain a variety of alkaloids. For example, Chinese herbs such as mountain bean root, nightshade, shegan, astragalus, licorice and other traditional Chinese medicines are often used in combination when prescribing. In addition, sprouted potatoes, immature tomatoes and eggplants, and immature nightshade fruits al...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72
CPCG01N30/02G01N30/06G01N30/30G01N30/32G01N30/34G01N30/72G01N2030/324
Inventor 赵金宝贾永娟刘鑫倪君君
Owner BEIJING HARMONY HEALTH MEDICAL DIAGNOSTICS CO LTD
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