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Methods for separation and content determination of chlorogenic acid type components in gynura procumbens

A technology of Pingwoju notoginseng and chlorogenic acid, applied in the quantitative determination of neochlorogenic acid and three kinds of isochlorogenic acids, chlorogenic acid in Pingwoju notoginseng extract, determination of Pingwoju notoginseng extract Quality inspection field

Active Publication Date: 2015-09-23
谭玉莲
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Problems solved by technology

The existing literature reports the extraction and purification process of chlorogenic acid, but the research on other similar components of chlorogenic acid in Pingwo chrysanthemum notoginseng has not been involved, and there is no relevant report on its quality standard. Therefore, the applicant separates Prepare each component, and establish HPLC method to determine the content of each component at the same time

Method used

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  • Methods for separation and content determination of chlorogenic acid type components in gynura procumbens
  • Methods for separation and content determination of chlorogenic acid type components in gynura procumbens
  • Methods for separation and content determination of chlorogenic acid type components in gynura procumbens

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] Example 1 Isolation and Identification of Chlorogenic Acids in Pingwo Jusanqi

[0015] 1. Preparation of Pingwoju Sanqi extract sample

[0016] 1.1 Extraction sample preparation Pingbu notoginseng is produced in Guangdong and identified by Mr. Zhang Bingkun of Wuhan Botanical Garden, Chinese Academy of Sciences. Take the dried stem (0.5kg) and crush it, add 5L of 80v / v% ethanol each time, reflux extraction at 85°C for 1h, extract 3 times in total, combine the extracts and concentrate and dry, the extract (density 1.40g / L) yield was 18.5%. Dissolve the extract in 50v / v% methanol (the ratio of extract to 50v / v% methanol is 0.1g: 1mL), and filter it with a 0.45μm microporous membrane for preparation.

[0017] The above experiment was repeated three times, and four batches of extracts were obtained in total, with batch numbers of 2012 / 6 / 14, 2012 / 6 / 20, 2012 / 6 / 21, and 2012 / 9 / 6 for subsequent experiments.

[0018] 1.2 Preparative chromatography and mass spectrometry conditi...

Embodiment 2

[0026] Example 2 The method for determining the content of chlorogenic acid, neochlorogenic acid and three kinds of isochlorogenic acids by external standard method

[0027] 2.1 Analytical chromatographic conditions

[0028] DIONEX UltiMate 3000 high performance liquid chromatograph; DIONEX C18 chromatographic column (250nm×4.6mm, 5μm), mobile phase is acetonitrile (A)-0.1w / v% phosphoric acid aqueous solution (B) (w / v unit is g / mL ), gradient elution: 0-65min, 9%-27%A (v / v%); 65-70min, 27%-9% (v / v%)A; time is 70min, volume flow 1.0mL / min , the injection volume was 10 μL, the detection wavelength was 326 nm, and the column temperature was 30°C.

[0029] 2.2 Solution preparation

[0030] 2.2.1 Sample preparation

[0031] Accurately weigh 100-250mg of the extract in Example 1, add 9mL of 50v / v% methanol to a 10mL volumetric flask, ultrasonicate at 20-40KHz for 15-30min in an ultrasonic instrument, let cool, add 50v / v% methanol to the mark Shake well and pass through a 0.45 μm m...

Embodiment 3 1

[0053] Embodiment 3 One test multiple evaluation method methodological investigation

[0054] 1 Calculation of relative correction factor On the basis of embodiment 2, calculate relative correction factor (f k / s ), calculation formula: f k / s =(C s ×A k ) / (C k ×A s ), the formula for calculating the mass concentration of the component to be tested: C k′ =(C s ×A k′ ) / (f k / s ×A s ), where C s is the concentration of the reference substance, A s is the chromatographic peak area of ​​the reference substance, C k is the mass concentration of other control groups, A k is the peak area of ​​other control chromatographic peaks, C k′ is the mass concentration of the component to be measured, A k′ is the area of ​​the chromatographic peak of the component to be tested, and the results are shown in Table 3.

[0055] Table 3 f with chlorogenic acid as reference k / s (multi-point calibration method)

[0056]

[0057] 2 Inspection of the reproducibility of the correction ...

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Abstract

The invention discloses two methods for quantitative determination of chlorogenic acid, neochlorogenic acid and three isochlorogenic acids in gynura procumbens extract. The two methods comprise the steps that 1, the best chromatogram conditions are determined; 2, reference substances of the neochlorogenic acid, the chlorogenic acid and the three isochlorogenic acids are obtained, methyl alcohol is added to the reference substances, and then a reference solution is prepared; 3, the gynura procumbens extract is obtained, methyl alcohol is added to the gynura procumbens extract, and after filtering, a sample is obtained; 4, the reference solution and the sample solution are precisely absorbed, chromatography sample introduction is conducted, and the contents of the chlorogenic acid, the neochlorogenic acid and the three isochlorogenic acids are measured; 5, or, only a chlorogenic acid reference solution is prepared in the step 2, the chlorogenic acid is used as a reference substance, the relative correction factors of the isochlorogenic acid C, the isochlorogenic acid A, the isochlorogenic acid B and the neochlorogenic acid are 1.21, 1.12, 1.07 and 0.92, and the contents of the five substances are measured. According to the two methods, the chlorogenic acid is used as reference, fk / s between the chlorogenic acid and other components is established, the content of each component is calculated, the external standard method and the method for quantitative analysis of multi-components by a single marker are similar in accuracy and reliability, and therefore a brand-new mode is provided for evaluating the quality of gynura procumbens more authentically.

Description

technical field [0001] The present invention relates to the field of quality control of Chinese herbal medicines, in particular to a quality detection method for Pingwo Jusanqi extract, more specifically, to the identification of chlorogenic acid, neochlorogenic acid and three kinds of isochlorogenic acids in Pingwo Jusanqi extract Quantitative determination method. Background technique [0002] Gynura procumbens (Lour.) Merr, commonly known as Pa Bengban, is a perennial herb of the genus Gynura procumbens (Lour.) Merr. It is used as medicine and food, and has a wide range of pharmacological effects. Blood stasis, swelling, blood circulation and muscle growth", in Southeast Asian ethnic areas around my country, it is also used for lowering blood sugar and lipids, resisting liver cancer, detoxifying viscera and protecting the intestinal tract. The folks often use the tender stems and leaves as vegetable food and tea drinks as new food raw materials (former Ministry of Health ...

Claims

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Application Information

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IPC IPC(8): G01N30/02
Inventor 李小军唐和斌张妹砣李婷婷杨艳羚穆云妹李玉桑
Owner 谭玉莲
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