High efficiency liquid chromatography method for simultaneously quantitatively detecting six flavonoid components in polygonum hydropiper

A technology of high performance liquid chromatography and detection method, which is applied in the field of high performance liquid chromatography for simultaneous quantitative detection of six flavonoids in Polygonum serrata, can solve the problems of high separation efficiency, expensive equipment and inability to detect the total flavonoid content of traditional Chinese medicine, etc. problem, to achieve the effect of improving the detection efficiency

Active Publication Date: 2014-04-30
INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The HPLC method has relatively less interference, high separation efficiency, high sensitivity, good reproducibility, and more accurate and reliabl

Method used

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  • High efficiency liquid chromatography method for simultaneously quantitatively detecting six flavonoid components in polygonum hydropiper
  • High efficiency liquid chromatography method for simultaneously quantitatively detecting six flavonoid components in polygonum hydropiper
  • High efficiency liquid chromatography method for simultaneously quantitatively detecting six flavonoid components in polygonum hydropiper

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] The validity test of embodiment 1 method

[0017] Preparation of reference substances: Precisely weigh 10 mg each of rutin, hyperin, quercitrin, quercetin, isorhamnetin, and kaempferol reference substances into 10ml volumetric flasks, dilute to 10ml with methanol, Shake well, prepare 1mg / ml stock solution, and set aside. Take 0.5ml of six kinds of stock solutions and place them in 25ml volumetric flasks to obtain a mixed standard solution of 20.0ug / ml, and then dilute to obtain mixed standard solutions of 10.0, 5.0, 1.0, and 0.1ug / ml. Preparation of the test sample: Grind Polygonum syringae, pass through a 20-mesh sieve, add 10 times the amount of methanol, weigh and record the data, after ultrasonication for 40 minutes, weigh and make up to the previous weight with methanol, filter, and take the filtrate for later use. Use high-performance liquid chromatography ultraviolet detector to detect at optimum wavelength 359nm place, the chromatogram of 10ug / ml mixed refe...

Embodiment 2

[0034] Example 2 Addition recovery experiment to test the effectiveness of the method.

[0035] Accurately weigh 1.00 g of 5 parts of known content of Polygonum polygonum powder. Add 1.0mg of rutin, 0.5mg of hyperoside, 0.17mg of quercetin, 0.5mg of quercetin, 0.05mg of isorhamnetin, and 0.5mg of kaempferol in each serving, and extract according to the operation under "linear relationship" . Recovery %=(actually measured value-amount of analyte contained in the test product) / amount of reference substance added*100%. Described result is shown in table 1~table 6.

[0036] Table 1 Determination results of the recovery rate of rutin in Polygonum hydroponica

[0037]

[0038] Table 2 Determination results of hyperin recovery rate in Polygonum hydropicum

[0039]

[0040] Table 3 The results of the determination of the recovery rate of quercetin in Polygonum hydroponica

[0041]

[0042] Table 4 Determination results of recovery rate of quercetin in Polygonum waterpi...

Embodiment 3

[0049] Embodiment 3 Method application of the present invention

[0050] Adopt the method of the present invention, carry out the mensuration of flavonoid component content to the Polygonum polygonum of 7 different origins and batches, and mensuration result is as shown in table 7:

[0051] Table 7 Determination results of six flavonoids in Polygonum pepper samples

[0052]

[0053] Note: "—" means the substance was not detected.

[0054] According to the test results in Table 7, the production Figure 4 , the results of analysis and testing showed that the contents of the six substances in different Polygonum sp. Generally speaking, Polygonum pepper mainly contains three flavonoids, rutin, hyperin and quercetin, and different origins and picking months will affect their content in medicinal materials; while the contents of kaempferol and isorhamnetin are relatively low , the relevant components could not even be detected in individual producing areas; the difference in ...

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Abstract

The invention discloses a high efficiency liquid chromatography method for simultaneously quantitatively detecting six flavonoid components in polygonum hydropiper. The method comprises the following steps of selecting a Waters chromatographic column, selecting methanol and a 0.4% formic acid water solution as flowing phases, performing gradient elution, wherein the flowing speed of the flowing phase is 1ml/min, the gradient elution is finished in 3 gradients, and the volume ratios of methanol and 0.4% formic acid water solution of 3 gradients are respectively 45:55, 55:45 and 70:30, respectively performing ultraviolet detection on eluates at 359 nm, and determining the concentration of each component. The method provided by the invention has the advantages that the method is exact and simple and convenient, the replicability is good, the method is used for measuring the contents of rutin, hyperin, quercetin, quercetinic acid, isorhamnetin and kaempferol in the polygonum hydropiper, and the reference is provided for controlling quality of a polygonum hydropiper drug.

Description

technical field [0001] The invention relates to the field of flavonoid detection, and more specifically, relates to a high-performance liquid chromatography method for quantitatively detecting six flavonoid components in Polygonum hydroponica simultaneously. Background technique [0002] Polygonum hydropiper Linn. is the whole herb of Polygonum hydropiper Linn., also known as willow grass, Polygonum hydropiper Linn., Banana grass, Bat grass, etc. The whole plant can be used as medicine, hot and spicy Polygonum tastes pungent and warm; it has the effects of dispelling wind and dampness, dissipating stasis and relieving pain, detoxifying and reducing swelling, killing insects and relieving itching; it is mainly used for dysentery, gastroenteritis, diarrhea, rheumatic joint pain, Swelling and pain, functional uterine bleeding; external use to treat snake bites, skin eczema and other symptoms. [0003] There are many types of flavonoids, and their respective efficacies are also...

Claims

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Application Information

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IPC IPC(8): G01N30/06G01N30/60G01N30/02
Inventor 彭新宇何立美陈玉婷刘洪梅袁明贵尹烨华罗胜军魏光伟
Owner INST OF ANIMAL HEALTH GUANGDONG ACADEMY OF AGRI SCI
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