TUM. Biocompounds II Concentration of Flavonols and Anthocyanidins in apple skin: Relation between Multiplex and HPLC values. Prof. Dr.

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1 TUM Biocompounds II Concentration of Flavonols and Anthocyanidins in apple skin: Relation between Multiplex and HPLC values Prof. Dr. Treutter Dr. Rühmann Michele Lomonaco

2 Flavonols Are derived from dihydroflavonols by the flavonol synthase (FLS).Are a yellow class of flavonoids. Belong to flavonols: quercetin, kaempferol (fig.1), myricetin. Fig. 1 Kaempferol Anthocyanidin Are synthesised from flavan 3,4-diols (leucoanthocyanidins) by the enzyme anthocyanidin synthase (ANS). Belong to flavonols: Pelargonidin (Fig.2), Cyanidin, Delphinidin. Fig. 2 Pelargonidin Comparison between HPLC and Multiplex MULTIPLEX By using this method we can measure the phenolics in epidermis cells. We can measure directly in the tree. But this method was adjusted for anthocyanidines and flavonols (two phenolics groups). With this method we only get one term saying something about a total concentration of a lot compounds. When anthocyanins are present in the epidermis they absorb green light and screen the mesophyll. Flavonoids are present in the epidermis. They absorb UV radiation and screen the mesophyll.

3 HPLC High-performance liquid chromatography (or High pressure liquid chromatography, HPLC) is a form of column chromatography used to separate, identify, and quantify compounds.hplc utilizes a column that holds chromatographic packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.retention time varies depending on the interactions between the stationary phase, the molecules being analyzed, and the solvent(s) used. Work description In this work two cultivars were chosen: Braeburn (6 apples) and Royal Gala (5 apples), on both cultivars were measured the concentrations of flavonols and Anthocyanidins, either by direct method (multiplex) that with the destructive method ( HPLC).

4 RESULTS AND DISCUSSION Multiplex index in Braeburn flavonols anthoc Sun dark Sides This graph represents the amount of flavonols and anthocyanidins in Braeburn, measured with the multiplex, and we can see how the amount of both compounds is higher in the parts most affected by the sun (1 and 2).

5 HPLC concentrations in Braeburn Also this graphs represent the amount of flavonols and anthocyanidins in Braeburn,but, measured with the multiplex, and we can see, how also in this case, the amount of both compounds is higher in the parts most affected by the sun (1 and 2).

6 Graphs of the correlations between the measurements made with the mutiplex and those carried out by HPLC GALA - ANTHOCYANIDINS GALA - FLAVONOLS In these two graphs above, we may notice the lack of correlation between the measurements made by the direct method (multiplex) and the destructive method (HPLC), the amount of flavonols and Anthocyanidins on cultivar Gala.

7 BRAEBURN ANTHOCYANIDINS BRAEBURN FLAVONOLS Also in these two graphs above, we may notice a low correlation between the measurements made by the direct method (multiplex) and the destructive method (HPLC), the amount of flavonols and Anthocyanidins on cultivar Braeburn.

8 COX ORANGE ANTHOCYANIDINS COX ORANGE FLAVONOLS In the cultivars Cox Orange, the correlation for the anthocyanidins is about 's 80%, while for the correlation is falvonoli' s 88%. Both correlations are quite high.

9 GRANNY SMITH ANTHOCYANIDINS GRANNY SMITH FLAVONOLS In the cultivars Granny Smith, the correlation for the anthocyanidins is 4.7% lower. Perhaps the reason is that the concentration of anthocyanidins in this cultivar is very low. While, for flavonoli the correlation is 73.7%.

10 CANDLE ANTHOCYANIDINS CANDLE FLAVONOLS Candle In cultivars, the correlation for the anthocyanidins is 76.3%, while for the correlation is falvonoli 's 82.7%. A good correlation.

11 ELSTAR ANTHOCYANIDINS ELSTAR FLAVONOLS In the cultivar Elstar, the correlation for the anthocyanidins is 47.2%, very low. For the correlation is falvonoli 's 85.8% (good correlation).