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High performance liquid chromatography (HPLC) quantitative determination method for natural caffeine in guarana fruit powder

A technology of high performance liquid chromatography and guarana fruit, applied in the field of determination of caffeine content, to achieve the effect of accurate determination, good linear relationship and good repeatability

Inactive Publication Date: 2012-07-04
北京知蜂堂健康科技股份有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0010] At present, there are few reports in the literature about the analysis method of caffeine in guarana fruit powder, and there are few studies on this at present. Therefore, it is very important to study a method that can accurately measure the caffeine content in guarana fruit powder

Method used

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  • High performance liquid chromatography (HPLC) quantitative determination method for natural caffeine in guarana fruit powder
  • High performance liquid chromatography (HPLC) quantitative determination method for natural caffeine in guarana fruit powder
  • High performance liquid chromatography (HPLC) quantitative determination method for natural caffeine in guarana fruit powder

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Experimental program
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Effect test

Embodiment 1

[0063] Prepare caffeine standard solution according to the method described above, after the standard solution obtained above is diluted five times with dehydrated alcohol, inject liquid chromatograph after filtering through 0.45 μm microporous membrane, according to the method determined by the present invention, in the following The determination was carried out under the stated chromatographic conditions.

[0064] The results show that the peaks are good, and the chromatographic peak of caffeine is completely separated from other component chromatographic peaks, such as figure 1 shown.

[0065] Wherein, chromatographic column: Kromasil C18 (250mm * 4.6mm, 5 μ m); Mobile phase: acetic acid: methanol: DMF: water (2: 3: 35: 160, volume ratio); Chromatographic column temperature: 25 ℃; Injection volume: 5 μl; flow rate: 0.9mg / ml; detector: ultraviolet detector; detection wavelength: 280nm.

Embodiment 2

[0067] Prepare 6 sample solutions of different batch numbers according to the above-mentioned method, after the sample solution obtained above is diluted five times with absolute ethanol, inject the liquid chromatograph after filtering through a 0.45 μm microporous membrane, and determine according to the method of the present invention , determined under the chromatographic conditions described below.

[0068] Each sample is injected once, and the chromatographic peak of caffeine in the sample can be determined through the standard spectrum of caffeine. Record the peak area, calculate the caffeine content of each sample according to the external standard method, the results are: 4.45%, 4.36%, 4.50%, 4.72%, 4.47%, 4.36%, and the sample under this condition, the caffeine contained in the sample Because it can achieve baseline separation with other impurities, its chromatographic peak shape is good, such as figure 2 shown.

[0069] Wherein, chromatographic column: Kromasil C1...

Embodiment 3

[0071] Prepare caffeine standard solution and guarana fruit powder sample solution according to the method described above, according to the method determined by the present invention, after the solution (standard solution and sample solution) obtained above is diluted five times with dehydrated alcohol, pass through 0.45 μm micro After filtering through a porous filter membrane, inject into a liquid chromatograph, and measure according to the method of the present invention, it can be obtained that the content of natural caffeine in the guarana fruit powder is 4.55%.

[0072] Wherein, chromatographic column: Kromasil C18 (250mm * 4.6mm, 5 μ m); Mobile phase: acetic acid: methanol: DMF: water (2: 3: 35: 160, volume ratio); Chromatographic column temperature: 25 ℃; Injection volume: 5 μl; flow rate: 0.9mg / ml; detector: ultraviolet detector; detection wavelength: 280nm.

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Abstract

The invention relates to a high performance liquid chromatography (HPLC) quantitative determination method for natural caffeine in guarana fruit powder. According to the method, the caffeine content in the guarana fruit powder is determined by using an external standard method. Liquid chromatography conditions are that: C18, C8, C12 or phenyl chromatographic column is adopted; a mixed solvent constituted by ethanol, methanol, DMF (Dimethyl Formamide) and water is used as a mobile phase; the volume ratio of the ethanol to the methanol to the DMF to the water is 1-3:2-4:30-38:150-170; and the detection wavelength is 280 nm. By using the method, the linear range of the caffeine is quantitatively determined to be 0.57-11.4 micrograms, the linear correlation coefficient r is equal to 0.9996, the sample standard average recovery rate is 100.36 percent, and RSD (Relative Standard Deviation) is 0.39 percent. The method is easy and convenient to operate, has high repeatability, is fast, sensitive, accurate and believable, and can be directly used for determining content of the natural caffeine in the guarana fruit powder in a production process.

Description

technical field [0001] The invention relates to a method for measuring caffeine content, in particular to a method for quantitatively measuring natural caffeine in guarana fruit powder by high performance liquid chromatography (HPLC). Background technique [0002] Guarana is a wild berry native to the Amazon region. It contains natural green caffeine, vitamins, and rich alkaloids, tannins, and proteins. The beverage made by using it has unique flavor and is also a healthy beverage. Guarana is a transliteration of the English name guarana. It is a woody vine of Sapindaceae, and the medicinal part is a seed. It is produced in the tropical forest of the Amazon River Basin in Brazil and is a unique economic crop in the Amazon region. The natural caffeine it contains is 2-4.5 times that of coffee beans. It can gradually and slowly stimulate the nerves. The stimulation lasts for a long time, and the stimulation is relatively mild. 3. The stimulating effect of tea on nerves usua...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/89
Inventor 李育强李楠刘枫黄冀东许文舫
Owner 北京知蜂堂健康科技股份有限公司
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