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

A technology for active ingredients and content, applied in the field of simultaneous determination of the content of various active ingredients in Eucommia ulmoides, can solve the problems of weakened absorption peak intensity, poor separation of chromatographic peaks, and difficulty in simultaneous determination, etc., to achieve accurate determination methods and improve detection. The effect of efficiency

Inactive Publication Date: 2013-01-23
CENTRAL SOUTH UNIVERSITY OF FORESTRY AND TECHNOLOGY
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  • Summary
  • 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

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Method verification experiment

[0025] 1 Investigation of linear relationship

[0026] Accurately weigh the pinoresinol diglucoside, geniposide acid, chlorogenic acid, and rutin reference substance, and dissolve it with methanol to make the concentration 0.11mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 , Mix the reference solution, and prepare solutions of different series of concentrations into the high performance liquid chromatograph, and measure the peak area according to the above chromatographic conditions (see the chromatogram figure 1 ), with the injection volume of each reference substance (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 the peak area of ​​each reference substance has a good linear relationship with the concentration.

[0027] Table 3 Results of linear relationship investigation

[0028]

[0029]

[0030] 3. Precision experiment

[0031] Precisely d...

Embodiment 2

[0039] Example 2 Test of active ingredients in Eucommia bark

[0040] The octadecylsilane bonded silica gel is used as a filler; the mobile phase A is methanol, and the mobile phase B is an aqueous solution of phosphoric acid with a mass concentration of 0.1%. 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℃; Injection volume is 5μl.

[0041] Preparation of reference solution: accurately weigh pinoresinol diglucoside, geniposide acid, chlorogenic acid, and rutin reference substance, and dissolve it with methanol to make the concentration 0.11mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 The mixed reference solution is injected with the resolution not less than 1.5 and the theoretical plate is not less than 5000.

[0042] Preparation of test solution: accurately weigh 5 g of Eucommia ulmoides skin after 40 mesh sieve, add 50 mL of methanol in a round bottom flask, and ex...

Embodiment 3

[0043] Example 3 Test of active ingredients in Eucommia ulmoides leaves

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

[0045] Preparation of reference solution: accurately weigh pinoresinol diglucoside, geniposide acid, chlorogenic acid, and rutin reference substance, and dissolve it with methanol to make the concentration 0.11mg·mL -1 , 0.11mg·mL -1 , 0.2mg·mL -1 , 0.1mg·mL -1 The mixed reference solution is injected with the resolution not less than 1.5 and the theoretical plate is not less than 5000.

[0046] Preparation of test solution: accurately weigh 2g of Eucommia ulmoides leaves after 40-mesh sieve, add 30mL methanol in a round-bottomed flask, and extract for 10min i...

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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, in particular to the simultaneous determination of geniposide acid (GPA), chlorogenic acid (CA), pinoresinol diglucoside (PDG) and rutin (Rutin) Method for determination of content. Background technique [0002] Eucommia ulmoides OLiv (Eucommia ulmoides OLiv) is a perennial deciduous tree belonging to Eucommia ulmoides family. 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 invigorating liver and kidney, strengthening muscles and bones, restoring fetus, and lowering blood pressure. The main active ingredients are lignans, iridoids, phenylpropanoids, flavonoids and so on. The main representative components are pi...

Claims

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Application Information

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Patent Type & Authority Applications(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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