HPLC (High Performance Liquid Chromatography) detection method for simultaneously determining 11 kinds of flavonoids and method for detecting content of flavonoids in fruits

A high-performance liquid chromatography and detection method technology, applied in the field of flavonoid content detection, can solve the problems of cumbersome work, large differences in detection conditions and methods, and large quantities

Active Publication Date: 2018-01-09
POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

High performance liquid chromatography is simple to operate, high in accuracy and good in reproducibility. However, because the detection conditions and methods of each component of anthocyanins, flavonols and flavonoids are quite different, multiple detections in batches are often required. The work is cumbersome and heavy

Method used

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  • HPLC (High Performance Liquid Chromatography) detection method for simultaneously determining 11 kinds of flavonoids and method for detecting content of flavonoids in fruits
  • HPLC (High Performance Liquid Chromatography) detection method for simultaneously determining 11 kinds of flavonoids and method for detecting content of flavonoids in fruits
  • HPLC (High Performance Liquid Chromatography) detection method for simultaneously determining 11 kinds of flavonoids and method for detecting content of flavonoids in fruits

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0108] Respectively weigh delphinidin glucoside, delphinidin rutinoside, paeoniflorin glucoside, cyanidin rutinoside, cyanidin glucoside, rutin, and cyanidin glucoside, which are HPLC chromatographically pure (purity ≥ 99%). Propylene glycol, quercetin, myricetin, isorhamnetin, apigenin reagent, methanol-hydrochloric acid (V 甲醇:V 盐酸 =98:2) was dissolved and prepared into a mixed standard solution with a concentration of 2.5, 5.0, 50.0, 100.0, and 500.0 μg / mL, and was filtered with a 0.22 μm organic filter membrane for HPLC analysis.

[0109] The mixed standard solution is subjected to high performance liquid chromatography analysis, and the chromatographic conditions are:

[0110] Analytical column: C-18 column, InfinityLab Poroshell 120EC-C18, 4.6×150mm;

[0111] Mobile phase: water phase A: 0.3wt% phosphoric acid aqueous solution; organic phase B: acetonitrile;

[0112] Flow rate: 1mL / min;

[0113] Elution method: Gradient elution, 0min, A:B=95:5, 20min, A:B=85:15, 30min...

Embodiment 2

[0125] Get black gem plum fruit peel 5g, liquid nitrogen grinds to pulverization, 25ml methanol-hydrochloric acid solution (V 甲醇 :V 盐酸 =99:1) leached in the dark for 12h, filtered; the filter residue was re-used with 25ml methanol-hydrochloric acid solution (V 甲醇 :V 盐酸 =99:1) repeated leaching 2 times; all filtrates were collected with methanol-hydrochloric acid solution (V 甲醇 :V 盐酸 =99:1) was fixed to 100ml to obtain a sample solution, the sample solution was centrifuged at 8000rpm at 4°C for 10min, and then filtered with a 0.22μm organic filter membrane to obtain a test solution for future use.

[0126] Adopt the same high performance liquid chromatography detection condition as embodiment 1 to detect the liquid to be tested, the high performance liquid chromatography spectrum that obtains is as follows figure 2 as shown, figure 2 Among them, 1 is delphinidin glucoside, 2 is delphinidin rutinoside, 3 is cyanidin glucoside, 4 is cyanidin rutinoside, 5 is rutin, 6 is my...

Embodiment 3

[0132] Using the same steps as in Example 2, the only difference is that the black gem plum peel is replaced with cherries (peel and pulp). The obtained HPLC chromatogram is as follows image 3 as shown, image 3 Among them, 1 is cyanidin glucoside, 2 is cyanidin rutinoside, and 3 is rutin.

[0133] According to the standard equation that embodiment 1 provides, calculate the content of different types of flavonoids in the black gem plum peel, the results are as shown in table 3, by image 3 As can be seen from Table 3, Example 3 achieves the determination of flavonoids in cherries through one detection.

[0134] Table 3 Flavonoid content in cherries (μg / g·FW)

[0135]

[0136] Wherein, FW is the fresh weight (g) of the fresh fruit sample. In addition, it should be noted that in the analysis of the results of the above-mentioned examples, only the content of flavonoids that can be detected is given, and the content is less than 5 μg / g·FW or not The detected ones are not ...

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Abstract

The invention provides an HPLC (High Performance Liquid Chromatography) detection method for simultaneously determining 11 kinds of flavonoids, and belongs to the field of flavonoids detection. The 11kinds of flavonoids are delphinidin glucoside, delphinidin rutinoside, peonidin glucoside, cyanidin rutinoside, cyanidin glucoside, rutin, kaempferol, quercetin, myricetin, isorhamnetin and apigenin.According to the HPLC detection method provided by the invention, different kinds of flavonoids matters can be detected within different retention times through accurate limit on an HPLC condition, the minimum separation degree of a chromatographic peak is 1.54, and simultaneous determination of 11 kinds of flavonoids is realized; data of the embodiment shows that simultaneous determination of 11kinds of flavonoids is realized, and the operation is simple and quick; meanwhile, the standard deviation of the peak area and the retention time is smaller, and higher accuracy is realized.

Description

technical field [0001] The invention relates to the technical field of flavonoid content detection, in particular to a high-performance liquid chromatography detection method for simultaneously measuring 11 flavonoids and a method for detecting flavonoid content in fruits. Background technique [0002] Flavonoids, also known as bioflavonoids, are the most abundant polyphenolic compounds in the human diet. They are widely found in fruits, vegetables and other crops. At present, more than 4,000 different flavonoids have been confirmed, and the content in fruits is relatively high. The most flavonoids are flavonoids, flavonols and anthocyanins. Flavonoids have strong antioxidant and free radical scavenging capabilities, and have important biological activities such as regulating the cardiovascular system, endocrine system, anti-cancer, anti-cancer, and anti-aging. [0003] The analysis methods of flavonoids include ultraviolet spectrophotometry, aluminum nitrate colorimetry, h...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/14
Inventor 周丹蓉叶新福方智振潘少霖姜翠翠
Owner POMOLOGY RES INST FUJIAN ACAD OF AGRI SCI
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