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
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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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