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Method for determining content of three antioxidation active compositions in honeysuckle flower and plant thereof by employing HPLC

A technology of antioxidant activity and honeysuckle, which is applied in the fields of food, biomedicine, and health products, can solve the problems of unfavorable rapid analysis and mass sample quality control, large structural differences, and long measurement time, so as to shorten the detection workflow, High precision and good peak shape

Inactive Publication Date: 2016-04-13
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chlorogenic acid and luteolin are the representative medicinal ingredients in the medicinal material honeysuckle. The 2000 edition of the Chinese Pharmacopoeia published a method for the determination of chlorogenic acid content in honeysuckle by high performance liquid chromatography. Chromatography (HPLC method) is a standard method for determining the content of luteolin. However, due to the large difference in the molecular structure of chlorogenic acid and luteolin, different HPLC analysis methods are used to determine the content of the two substances separately. Two different chromatographic columns and detection conditions, the workload is heavy, the measurement time is long, which is not conducive to the needs of rapid analysis and mass sample quality control

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The method for simultaneously measuring the content of three antioxidant active ingredients in Flos Lonicerae and its plants by HPLC comprises the following steps:

[0027] 1. Preparation of the test sample solution: Take 0.5g of dried honeysuckle, weigh it accurately, cut it finely and put it in a stoppered Erlenmeyer flask, add 10m of ethanol with a mass concentration of 60%, ultrasonicate for 20min, pour out the supernatant, and then add 10ml of ethanol with a mass concentration of 60%, repeated ultrasonic extraction for 20min, combined the extracts, passed through a 0.45μm filter membrane, added ethanol to make the volume to 50ml, and obtained the honeysuckle test sample solution.

[0028] 2. Preparation of control sample solution: Accurately weigh a certain amount of reference substances of chlorogenic acid, luteolin, and luteolin, dissolve them fully in three beakers, and prepare them with ethanol containing 20 μg / mL of chlorogenic acid, Control solution of luteol...

Embodiment 2

[0042] The method for simultaneously measuring the content of three antioxidant active ingredients in Flos Lonicerae and its plants by HPLC comprises the following steps:

[0043] 1. Take 1.0 g of dried honeysuckle leaves, cut them finely and crush them, put them in a stoppered Erlenmeyer flask, add 15 ml of ethanol with a mass concentration of 70%, ultrasonicate for 40 minutes, repeat the ultrasonic extraction twice, combine the extracts, pass 0.45 μm filter membrane, add ethanol to make the volume to 50ml, and obtain the honeysuckle test sample solution.

[0044] 2. Preparation of control sample solution: Accurately weigh 5.0 mg of chlorogenic acid, 6.0 mg of luteolin, and 6.0 mg of luteolin as reference substances, dissolve them fully in three beakers, and use ethanol to make 50 μg / mL, luteolin 30 μg / mL, luteolin 30 μg / mL control sample solution.

[0045] 3. Determination of the content of chlorogenic acid, luteolin and luteolin in the leaves of honeysuckle leaves, using h...

Embodiment 3

[0057] The method for simultaneously measuring the content of three antioxidant active ingredients in Flos Lonicerae and its plants by HPLC comprises the following steps:

[0058] 1. Take 1.5g of dried honeysuckle leaves, cut them finely and crush them, put them in a stoppered conical flask, add 20ml of ethanol with a mass concentration of 80%, ultrasonicate for 30 minutes, repeat the ultrasonic extraction twice, combine the extracts, pass 0.45 μm filter membrane, add ethanol to make the volume to 50ml, and obtain the honeysuckle test sample solution.

[0059] 2. Preparation of control sample solution: Accurately weigh 5.0 mg of chlorogenic acid, 6.0 mg of luteolin, and 6.0 mg of luteolin as reference substances, dissolve them fully in three beakers, and use ethanol to make 50 μg / mL, luteolin 30 μg / mL, luteolin 30 μg / mL control sample solution.

[0060] 3. Determination of the content of chlorogenic acid, luteolin and luteolin in the leaves of honeysuckle leaves, using high p...

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Abstract

The invention discloses a method for determining content of three antioxidation active compositions in honeysuckle flower and plant thereof by employing HPLC. The method comprises getting 0.5-2.0 g of dry honeysuckle flower and plant leaf and stem of honeysuckle flower, chopping up and pulverizing, putting in a container, adding ethanol, and performing ultrasonic processing, so as to obtain a test sample solution; getting the test sample solution, applying to an ODS chromatographic column, and performing gradient elution, wherein the eluent is a mixed solution of acetonitrile and a phosphoric acid aqueous solution, and the concentration gradient of acetonitrile in the mobile phase is changed along with time; employing the detection wave length of 320-340 nm in the stage from initial time t0 to time t5, and then transforming the wave length to be 345-365 nm for detecting, wherein t5=10-18 min; and successively determining the content of three effective compositions chlorogenic acid, luteolin and luteoloside in honeysuckle flower. The provided method for simultaneously determining content of the three antioxidation active compositions in honeysuckle flower and plant thereof by employing HPLC is high in sensitivity and precision and simple in work flow.

Description

technical field [0001] The invention relates to the fields of biomedicine, food, health care products, etc., in particular to an HPLC method for simultaneous determination of biologically active natural products, in particular to a wavelength conversion technology applied in HPLC for the simultaneous detection of Chlorogenic acid, luteolin, luteolin 3 kinds of antioxidant active ingredients. Background technique [0002] Honeysuckle is a traditional Chinese medicine in my country, which is the dry flower bud or the first flower of Lonicera Lonicera in the Lonicera family. It is sweet in taste and cold in nature. It has the functions of clearing heat and detoxifying, dispelling wind-heat, dredging throat, relieving summer heat and annoyance. . Honeysuckle mainly contains volatile oils, organic acids, flavonoids, triterpenoid saponins, iridoid glycosides and other bioactive components. Among them, flavonoids represented by luteolin and its metabolic precursor luteolin have ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02
Inventor 郑必胜陆建超扶雄刘瑞海颜盛繁
Owner SOUTH CHINA UNIV OF TECH
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