# Aromatic and Nutritional Composition of Edible Flowers of Garden Garlic and Wild Leek
## 2 Materials and Methods
### Experimental Methods  
For the analysis, the floral axes of 15 plants were collected. The plant material was vacuum-packed and frozen until processing and analysis, except for volatile components determination, in which case fresh flowers were used. In all cases, the entire corolla of the flowers was analyzed, once separated from the peduncle and sepals.  
Proximal composition in the flowers was determined using official methods [29]: first, moisture and dry matter readings were taken using the stove J.P. Selectam (Barcelona, Spain) at 70 \(\pm\) 0.1 \({}^{\circ}\)C until a constant weight (AOAC 984.25 method) was achieved; next, proteins were determined using the Kjeldahl method (AOAC 984.13) with the Foss Tecator 2100 Kjeltec distillation unit (Hillerod, Denmark); fat was determined using a Foss Soxhlet model ST243 SoxtecTM from LabtecTM line (Hillerod, Denmark) (AOAC 983.23 method); fiber was determined by quantifying the residue that persists after two successive hydrolyses, one acidic and the other alkaline (AOAC 991.43 method); and ashes were determined via calcination in a Carbolite CWF 1100 chamber furnace at 550 \({}^{\circ}\)C (AOAC 923.03 method). The carbohydrate content was calculated using difference. The total energy was calculated by multiplying by 9 kcal the grams of fat, by 4 kcal the grams of protein and carbohydrates, and by 2 kcal the grams of fiber, each for 100 g of fresh flowers. The results are expressed as g\(\cdot\)100 g\({}^{-1}\) of dry weight (dw).  
The individual mineral composition in the flowers was carried out using inductively coupled plasma emission spectroscopy (ICP-EOS), from ashes dissolved and settled with concentrated HCl until a 2% HCl solution. The equipment used was Agilent ICP-EOS 710 (700 series ICP-OES), made in Mulgrave, Victoria, Australia. The wavelengths selected for each element were the following: 177.434 nm for P, 184.887 for Hg, 196.026 nm for Se, 213.857 nm for Zn, 228.802 nm for Cd, 238.204 nm for Fe, 249.678 nm for B, 257.610 nm for Mn, 267.716 nm for Cr, 281.615 nm for Mo, 285.213 nm for Mg, 317.933 nm for Ca, 324.754 nm for Cu, 405.781 nm for Pb, 589.592 nm for Na, and 769.897 nm for K. Results are expressed in mg of the mineral element per 100 g of dry flower.  
Antiradical activity via DPPH assay determination in flowers was performed according to the method described by [PERSON] et al. [30] based on the capture of the free radical DPPH. A methanolic extract was obtained by mixing 0.8 g of flowers in 5 mL methanol solution (80% \(v/v\)); it was stirred for 1 h at room temperature, using an orbital shaker (Stuart Scientific, Chelmsford Essex, UK). The DPPH solution (25 ppm in methanol 80% \(v/v\)) was prepared, mixing 3.9 mL of this solution with 0.1 mL methanolic extract. Absorbance was measured at 515 nm after 45 min of incubation with DPPH solution in the dark using a spectrophotometer (Schott UV line 9400, Essex, UK). The antioxidant Trolox was used as standard, and the results were expressed as micromoles of Trolox equivalents in each 100 g of dry weight (\(\mu\)mol TE\(\cdot\)100 g\({}^{-1}\) dw).  
Total polyphenols were determined according to the Folin-Ciocalteu procedure modified by [PERSON] et al. [31]. An aliquot (50 \(\mu\)L) of aqueous extract obtained by grinding the fresh flowers with distilled water in a ratio of 2:1 (solvent:flower) was mixed with 500 \(\mu\)L of the Folin-Ciocalteu reagent (previously diluted with water 1:10 \(v/v\)) and 500 \(\mu\)L of 6% (\(w/v\)) Na\({}_{2}\)CO\({}_{3}\) solution. The mixture was agitated for 10 s and allowed to stand for 1 h at room temperature for color development. The absorbance at 750 nm was measured usinga spectrophotometer (Jenway 6715/UV-V, Stone, Staffordshire, UK), using gallic acid as standard. The results were expressed as mg gallic acid equivalents per 100 g of dry weight (mg EAG 100 g\({}^{-1}\) dw).