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Method for simultaneously determining 5 alkaloids in Sini agent vegetable drug plasma

A botanical and plasma technology, applied in measurement devices, instruments, scientific instruments, etc., can solve difficult quality control, formulation design, clinical rational drug use, no simultaneous determination method, lack of pharmacokinetics of Sini Decoction botanicals, etc. problems, to achieve the effect of sensitive analytical methods

Inactive Publication Date: 2014-01-15
SHANGHAI ZHANGJIANG ENG RES CENT OF MODERN PREPARATION TECH OF TRADITIONAL CHINESE MEDICINE
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Problems solved by technology

[0002] Sini Decoction is composed of Fuzi, Dried Ginger, Radix Glycyrrhizae and other medicines, which have good preventive effects in protecting the myocardium, improving heart function, and preventing ischemia-reperfusion injury, etc. The study of metabolic kinetics is difficult to provide a basis for quality control, dosage form design, and clinical rational drug use
[0004] So far, the literature only has a method for the determination of dilipid diterpenoid alkaloids such as aconitine, neoaconitine, and hypoaconitine in plasma samples [He Leiping, Di Bin, Du Yingxiang, et al. Compatibility of four kinds of aconitine Pharmacokinetic comparison of three aconitum alkaloids in rat plasma after administration of prescription[J]. Journal of China Pharmaceutical University, 2010,41(1): 55-59; Shen Hong, Zhu Lingying, Yao Nan, et al .Effects of compatibility of licorice and aconitine on the pharmacokinetics of aconitine neoaconitine and hypoaconitine in rats[J].Chinese Materia Medica,2011,34(6):937-942], there is no four anti-drug plants Simultaneous determination method of aconitine, hypoaconitine, neoaconitine, benzoyl neoaconitine, and benzoyl aconitine in drug plasma samples

Method used

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  • Method for simultaneously determining 5 alkaloids in Sini agent vegetable drug plasma
  • Method for simultaneously determining 5 alkaloids in Sini agent vegetable drug plasma

Examples

Experimental program
Comparison scheme
Effect test

experiment example 1

[0015] Experimental example 1 Non-internal standard method linear relationship investigation

[0016] Take 7 parts of blank plasma, each 100 μl, add different concentrations of standard solution, so that the concentration of aconitine in the plasma is 0.06, 0.6, 3, 6, 30, 60, 300 ng·mL -1 , the concentrations of neoaconitine were 0.102, 1.02, 5.1, 10.2, 51, 102, 510 ng·mL -1 , Hypoaconitine 0.13, 1.3, 6.5, 13, 65, 130, 650 ng·mL -1 , the concentration of benzoyl neoaconitine is 0.1, 1, 5, 10, 50, 100, 500 ng·mL -1 , the concentrations of benzoylaconitine were 0.12, 1.2, 6, 12, 60, 120, 600 ng·mL -1 , to prepare a series of standard solutions.

[0017] Take the concentration of each reference substance as the abscissa and the peak area of ​​the reference substance as the ordinate, and use weighted least square regression (weight coefficient 1 / C) to obtain the regression equation of each reference substance. The regression curve results show that:

[0018] Aconitine at ...

experiment example 2

[0019] Experimental example 2 Investigation of linear relationship with internal standard method

[0020] Take 7 parts of blank plasma, each 100 μl, add different concentrations of standard solution, so that the concentration of aconitine in the plasma is 0.06, 0.6, 3, 6, 30, 60, 300 ng·mL -1 , the concentrations of neoaconitine were 0.102, 1.02, 5.1, 10.2, 51, 102, 510 ng·mL -1 , Hypoaconitine 0.13, 1.3, 6.5, 13, 65, 130, 650 ng·mL -1 , the concentration of benzoyl neoaconitine is 0.1, 1, 5, 10, 50, 100, 500 ng·mL -1 , the concentrations of benzoylaconitine were 0.12, 1.2, 6, 12, 60, 120, 600 ng·mL -1 , to prepare a series of standard solutions.

[0021] Take the ratio of the concentration of each reference substance to the concentration of the internal standard X as the abscissa, and take the peak area of ​​the reference substance and the percentage of the peak area of ​​the internal standard Y as the ordinate, and use Y to X for weighted least squares regression (wei...

experiment example 3

[0023] Experimental example 3 Precision and recovery investigation

[0024] Take 100 μl of blank plasma, add aconitine (internal standard) solution and different concentrations of reference solution, and prepare each containing aconitine, neoaconitine and hypoaconitine, benzoyl neoaconitine, Benzoyl aconitine 1, 10, 100 ng·mL -1 3 concentrations of standard drug-containing plasma, 5 samples were made for each concentration, and the methodological precision was determined.

[0025] Another blank plasma was taken to obtain the blank plasma supernatant, and then different amounts of standard solution and aconitin solution were added to obtain aconitine, neoaconitine, hypoaconitine, and benzoyl neoaconitine. Base, benzoyl neoaconitine 1, 10, 100 ng·mL -1 the control solution. Injection analysis, calculation of extraction recovery. See Table 1. It can be seen from the table that the precision and extraction recovery experimental results of aconitine, neoaconitine, hypoaconi...

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Abstract

The invention relates to a method for simultaneously determining 5 alkaloids in Sini agent vegetable drug plasma, which comprises the following steps: (1) taking medicated plasma of a mammal given Sini agent vegetable drugs, adding bulleyaconitine A (interior label), alkalifying with ammonia water, scrolling, adding ethyl acetate for extraction, scrolling, centrifugating, extracting the subnatant with ethyl acetate twice, merging the ethyl acetate layers, blow-drying with nitrogen, and dissolving the residue with a mobile phase; and (2) HPLC-MS-MS (high performance liquid chromatograph-mass spectrometer-mass spectrometer) determination: the mobile phases comprise a phase A acetonitrile-0.1% glacial acetic acid (1:99, v / v) and a phase B acetonitrile-water (99:1, v / v); the mass spectrometer conditions are as follows: electric spray ion source (ESI) and cation detection mode; and the detection ions are as follows: aconitine m / z 646.3->586.3, mesaconine m / z 632.3->572.3, hypaconitine m / z 616.3->556.3, benzoylmesaconine m / z 590.3->558.3, and benzoylaconine m / z 604.3->554.3. The method is suitable for pharmacokinetic research of aconitine, hypaconitine, mesaconine, benzoylmesaconine and benzoylaconine in Sini agent vegetable drugs in a mammal.

Description

technical field [0001] The invention belongs to the field of pharmacokinetics, in particular to the effect of aconitine, hypoaconitine, neoaconitine, benzoyl neoaconitine, and benzoyl aconitine in breast-feeding Determination method in animal plasma. Background technique [0002] Sini Decoction is composed of Fuzi, Dried Ginger, Radix Glycyrrhizae and other medicines, which have good preventive effects in protecting the myocardium, improving heart function, and preventing ischemia-reperfusion injury, etc. The study of metabolic kinetics is difficult to provide a basis for quality control, dosage form design, and clinical rational drug use. [0003] The medicinal ingredients in Sini Decoction are mainly alkaloids in aconite. The alkaloids in aconite can be divided into double-lipid alkaloids, mono-lipid alkaloids and aminoalcohol alkaloids. Among them, aconitine, neoaconitine, hypoaconitine, etc. are diterpenoid alkaloids of dilipid type, and monolipid type alkaloids such ...

Claims

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Application Information

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IPC IPC(8): G01N30/88
Inventor 洪燕龙冯怡阮克锋赵怀彬吴飞王优杰
Owner SHANGHAI ZHANGJIANG ENG RES CENT OF MODERN PREPARATION TECH OF TRADITIONAL CHINESE MEDICINE
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