Method for simultaneously measuring content of multiple active ingredients in eucommia ulmoides

A technology of active ingredients and content, which is applied in the field of simultaneous determination of multiple active ingredients in Eucommia, can solve the problems of weakened absorption peak intensity, poor separation effect of chromatographic peaks, and difficulty in simultaneous determination, so as to improve detection efficiency and determination method accurate effect

Inactive Publication Date: 2014-05-07
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In actual operation, we found that pinoresinol diglucoside has the maximum absorption at 277nm, and the intensity of the absorption peak at 254nm is greatly weakened; weakened, so it is difficult to determine the two simultaneously; and the polarity of chlorogenic acid and pinoresinol diglucoside is similar, and the separation effect of the chromatographic peaks using the conventional isocratic elution method is not good

Method used

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  • Method for simultaneously measuring content of multiple active ingredients in eucommia ulmoides
  • Method for simultaneously measuring content of multiple active ingredients in eucommia ulmoides
  • Method for simultaneously measuring content of multiple active ingredients in eucommia ulmoides

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Embodiment 1 method verification chemical experiment

[0025] 1 Linear relationship investigation

[0026] Accurately weigh an appropriate amount of pinoresinol diglucoside, geniposide, chlorogenic acid, and rutin reference substances, and dissolve them in methanol to prepare a concentration of 0.11 mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 , mix reference substance solution, and be prepared into the solution of different series concentration and inject high-performance liquid chromatograph, measure peak area by above-mentioned chromatographic condition (chromatogram sees figure 1 ), with each reference substance injection volume (μg) as the abscissa, and the peak area A as the ordinate, draw a standard curve, the results are shown in Table 3, the results show that each reference substance peak area and concentration are in a good linear relationship

[0027] Table 3 Linear relationship inspection results

[0028]

[0029]

[0030] 3. Precision expe...

Embodiment 2

[0039] The test of active ingredient in the bark of Eucommia ulmoides in embodiment 2

[0040] Octadecylsilane bonded silica gel is used as filler; mobile phase A is methanol, and mobile phase B is 0.1% phosphoric acid aqueous solution. The mobile phase gradient elution program and detection wavelength program are shown in Table 1 and Table 2: Flow rate: 0.8ml.min -1 ; Column temperature: 30°C; Injection volume: 5 μl.

[0041] Preparation of reference substance solution: Accurately weigh an appropriate amount of reference substance of pinoresinol diglucoside, genipinic acid, chlorogenic acid, and rutin, and dissolve it in methanol to prepare a concentration of 0.11 mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 Injection of the mixed reference substance solution, the resolution is not lower than 1.5, and the theoretical plate is not lower than 5000.

[0042] Preparation of the test solution: Accurately weigh 5g of Eucommia rawhide after passing through a 40 mesh siev...

Embodiment 3

[0043] The test of active ingredient in the leaf of Eucommia ulmoides of embodiment 3

[0044] Octadecylsilane bonded silica gel was used as filler; mobile phase A was methanol, and mobile phase B was 0.1% phosphoric acid aqueous solution. Mobile phase gradient elution program The detection wavelength program is shown in Table 1 and Table 2: Flow rate: 0.8ml.min -1 ; Column temperature: 30°C; Injection volume: 5 μl. The column temperature was 30°C.

[0045] Preparation of reference substance solution: Accurately weigh an appropriate amount of reference substance of pinoresinol diglucoside, genipinic acid, chlorogenic acid, and rutin, and dissolve it in methanol to prepare a concentration of 0.11 mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 Injection of the mixed reference substance solution, the resolution is not lower than 1.5, and the theoretical plate is not lower than 5000.

[0046] Preparation of the test solution: Accurately weigh 2 g of eucommia leaves afte...

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Abstract

The invention discloses a method for simultaneously measuring the content of multiple active ingredients in eucommia ulmoides. The method adopts high performance liquid chromatography for analysis; the chromatographic condition is taking octadecylsilane bonded silica as a filler; the mobile-phase gradient elution process is: the mobile phase A is methanol, and the mobile phase B is an aqueous solution of phosphoric acid with mass concentration of 0.1%; the content of the mobile phase B is increased from 30% to 70% by mass according to the first-order linear gradient in 0.01-30.00min, reduced to 30% in 30.00-30.01min and kept at 30% in 30.01-40.00min; the flow rate is 0.8ml / min; the column temperature is 30 DEG C; the sample injection volume is 5mul; the detection wavelength detection process is: the 254nm wavelength is selected in 0.01-7.00min, the 277nm wavelength is selected in 7.00-12.50min, and the 254nm wavelength is selected in 12.50-40.00min; and multiple active ingredients include geniposidic acid, chlorogenic acid, pinoresinol diglucoside and rutin. According to the invention, the method is verified by linear relation survey, stability test, precision test, reproducibility test and sample injection recycling test, and the analysis method is proved to be accurate and reliable.

Description

technical field [0001] The invention relates to a method for determining the content of multiple active ingredients in Eucommia ulmoides, especially for the simultaneous determination of geniposide (GPA), chlorogenic acid (CA), pinoresinol diglucoside (PDG) and rutin (Rutin) method for the determination of content. Background technique [0002] Eucommia ulmoides OLiv is a perennial deciduous tree belonging to Eucommia ulmoides. It is a traditional economic crop in my country. It is known as the "living fossil" of Quaternary glaciers. It is distributed in the middle reaches of the Yangtze River and southern provinces. It is cultivated in Henan, Shaanxi, Gansu and other places. . Eucommia has the effects of nourishing liver and kidney, strengthening bones and muscles, anti-abortion, and lowering blood pressure. Its main active ingredients are lignans, iridoids, phenylpropanoids, flavonoids, etc. The main representative components are pinoresinol diglucoside (lignans), genipo...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N30/02
Inventor 李湘洲张胜胡文彬贺义昌
Owner CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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