High performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae

A technology of grass flavonoids and Houttuynia cordata, which is applied in the field of HPLC determination of the content of Houttuynia cordata flavonoids, can solve the problems of affecting the efficacy of drugs, low reliability of quality control indicators of Houttuynia cordata, and unstable volatile oil components. To achieve the effect of improving the quality control level

Inactive Publication Date: 2018-03-30
CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology allows us to accurately determine if certain substances are found within Hypericum perforans or Quercus acutissima by testing different techniques such as extractive procedures and various types of solvents used during their preparation process. It provides technical benefits that make it possible to improve the accuracy and reliability of detectings on this plant material.

Problems solved by technology

This patented technical problem addressed in this patents relates to controlling the purity level (quality) of extract from houtuynyoside barkings called Cathayan yellowhorn flower with high reliability during production processes such as drying and storage conditions.

Method used

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  • High performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae
  • High performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae
  • High performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae

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Experimental program
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Effect test

Embodiment 1

[0038] Example 1. Detection method of the present invention

[0039] 1. Detection method

[0040] (1) Preparation of the test product: 0.1g of Houttuynia cordata powder through a No. 4 60 mesh sieve was accurately weighed, placed in a stoppered conical flask, 50% methanol 15mL was accurately added, weighed, ultrasonically extracted for 30 minutes, and left to cool , Weigh the weight, make up the weight with methanol, shake well, filter with a microporous membrane, take the filtrate for chromatographic determination, see the chromatogram figure 1 .

[0041] (2) Preparation of reference substance: Weigh accurately 10.26 mg of hyperoside reference substance and prepare a reference solution with a concentration of 102.6 ng / mL, and accurately weigh 10.87 mg of quercetin reference substance to a concentration of 108.7 ng. / mL of the reference solution. Take the reference solution for chromatographic determination, see the chromatogram figure 2 .

[0042] (3) Detected by HPLC, the chromat...

Embodiment 2

[0050] Example 2. Process parameter screening of the method of the present invention

[0051] 1 Investigation of chromatographic conditions

[0052] (1) Mobile phase

[0053] The elution effect of methanol-water, acetonitrile-water, acetonitrile-0.1% phosphoric acid water and the spectrum effect were investigated. The results showed that the elution effect of acetonitrile-0.1% phosphoric acid water was better, and the measured spectrum had a relatively stable baseline and chromatographic peaks. The shape is better, so acetonitrile-0.1% phosphoric acid water was finally selected as the mobile phase.

[0054] (2) Mobile phase gradient

[0055] The chromatographic conditions with different ratios of acetonitrile-0.1% phosphoric acid to water ratios of 22:78, 20:80, 18:82 were investigated, and the sample maps obtained by comparing different elution procedures ( image 3 , Figure 4 , Figure 5 ), the chromatographic peak with a gradient of 18:82 has high resolution, less impurities, stab...

Embodiment 3

[0090] Example 3. Methodological investigation of the method of the present invention

[0091] (1) Exclusive investigation

[0092] Investigate whether the fingerprint of Houttuynia cordata flavonoids established by this method can express the characteristics of this species, namely uniqueness. Investigate the blank solvent (50% methanol) spectrum, 50% methanol negative sample has no chromatographic peak at the corresponding retention time, no interference, and meets the requirements.

[0093] (2) Linear investigation

[0094] Take 10.26mg and 10.87mg of hyperoside and quercetin reference substance, respectively, and prepare the concentrations to 0.03072mg / mL, 0.0768mg / mL, 0.1536mg / mL, 0.2304mg / mL, 0.3072mg / mL and 0.04924mg respectively / mL, 0.1231mg / mL, 0.2462mg / mL, 0.3693mg / mL, 0.4924mg / mL of the reference substance solutions were measured and a standard curve was established. The results are shown in Table 7.

[0095] Table 7 Results of linear relationship investigation

[0096] ...

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Abstract

The invention discloses a high performance liquid chromatography (HPLC) method for detecting contents of flavonoid components in herba houttuyniae. The method comprises the following steps: (1) preparing a test solution, taking herba houttuyniae powder, extracting the herba houttuyniae powder with lower alcohol, and filtering to obtain the test solution; (2) preparing a reference solution, takingreference substances, i.e., hyperoside and quercitrin, mixing, adding the lower alcohol to prepare the mixed reference solution; (3) respectively injecting the test solution and the reference solutioninto a high performance liquid chromatographic instrument for detecting, wherein the chromatographic conditions are as follows: chromatographic column: C18 chromatographic column; mobile phases: a mobile phase A is acetonitrile, and a mobile phase B is 0.1% phosphoric acid aqueous solution; elution procedure: isocratic elution is carried out for 30 minutes, and the volume ratio of A to B is equalto (18-22) to (82-78); detection wavelength: 205nm; (4) calculating according to the detection result so as to obtain the contents of the hyperoside and the quercitrin in the herba houttuyniae. The detection method provided by the invention is accurate and reliable as well as simple and quick, and lays a foundation for improving the quality control level of the herba houttuyniae.

Description

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Claims

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Application Information

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Owner CHENGDU UNIV OF TRADITIONAL CHINESE MEDICINE
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