## 2 Materials and Methods
### Determination of Properties of Encapsulated Pumpkin Oil Capsules  
The moisture content of the powders was measured according to the AOAC Method [22]. The bulk density of the powders was measured by the standard method using a 61-400 apparent density tester (Coesfeld Materialest, Dortmund, Germany).  
The particle size and morphology of the powders were determined similarly as in the case of carbohydrates.  
The colour of the powders was determined using equipment for digital image analysis (DIA) [23]. The equipment was consisted of CCD (charge-coupled device) Nikon DXM-1200 colour camera, Kaiser RB 5004 HF-High Frequency Daylight Copy Light set with 4 \(\times\) 36 W fluorescent light tubes (colour temperature about 5400 K) (Kaiser Fototechnik GmbH and Co., KG, Buchen, Germany) and LUCIA (Laboratory Universal Computer Image Analysis) G v. 4.8 software. The results were expressed in the CIEL*a*b* colour model [24]. The total colour difference (\(\Delta\)E) was calculated based on the fallowing equation:  
\[\Delta\text{E}=\sqrt{\left(\Delta\text{L}*\right)^{2}+\left(\Delta\text{a}* \right)^{2}+\left(\Delta\text{b}*\right)^{2}} \tag{2}\]  
The whiteness index (WI) was calculated based on the following equation:  
\[\text{WI}=100-\sqrt{\left(100-\text{L}*\right)^{2}+\left(\text{a}*\right)^{2} +\left(\text{b}*\right)^{2}} \tag{3}\]  
Oxidative stability (OSI) of the powders was evaluated by a 743 Rancimat (Metrom, Switzerland) eight-channel oxidative stability instrument. 2.5 g of each sample was weighed into a reaction vessel. The capped vessel was placed in a thermostated electric heating block. The temperature was set at 110 \({}^{\circ}\)C and an air flow rate of 20 L/h was applied. The induction time (expressed as hours) was measured for each sample.  
Determination of Content of Bioactive Unsaponifiable Compounds in Native and Encapsulated Pumpkin Oil  
The content of tocopherols was determined according to the method described by [PERSON] et al. [25]. The powder was diluted in n-hexane and centrifuged (11,500\(\times\)\(g\) for 10 min) in a 5417R-type Eppendorf centrifuge (Eppendorf AG). The tocopherols were analysed using an Agilent Technologies 1200 RP-HPLC apparatus with a fluorescence detector (Santa Clara, CA, USA). Separation was performed on a LiChrospher Si60 column (Merck, Darmstadt, Germany). The mobile phase was 0.7% iso-propanol in a n-hexane solution and flow rate was at 1 mL/min. The fluorescence detector was set at excitation and emission wavelengths of 296 nm and 330 nm, respectively. The content of tocopherols was calculated based on the external calibration curves.  
The content of sterols and squalene was determined according to the method described by [PERSON] et al. [26]. The sample was dissolved in n-hexane with 5\(\alpha\)-cholestane solution as an internal standard. The mixture was saponified in 2 M KOH at temperature 70 \({}^{\circ}\)C for 30 min. Sterols and squalene were extracted with diethyl ether. The dry extract was re-dissolved in pyridine and N,O-bis (trimethylsilyl) trifluoracetamide (BSTFA) with 1% trimethylchlorosilane (TMCS), and derivatization was conducted at 60 \({}^{\circ}\)C for 60 min. The mixture dissolved in heptane was analysed using GC-MS QP2010 PLUS system (Shimadzu, Kyoto, Japan). Separation was performed on a ZB-5 ms capillary column (Phenomenex, Torrance, CA, USA). Helium was used as a carrier gas. The temperatures were: injector 230 \({}^{\circ}\)C, the column in range from 70 \({}^{\circ}\)C to 310 \({}^{\circ}\)C, the interface of GC-MS 240 \({}^{\circ}\)C, the ion source 220 \({}^{\circ}\)C, and the electron energy was 70 eV. The content of sterols and squalene was calculated based on the internal standard method.