A simple calculation on dry basis may also overestimate the retention of these compounds (De Sá and Rodriguez-Amaya, 2004 and Rodriguez-Amaya, 1999). Therefore, besides the results being expressed as μg/g of sample, they were also presented based on the mass of raw food, multiplying the concentration obtained for the sample by the ratio of the food mass after processing and of the food mass prior to processing. Therefore, true Alectinib retention (% TR) was calculated by the equation proposed by Murphy, Criner and Gray (1975) cited by De Sá and Rodriguez-Amaya (2004), as follows: % TR = 100 × (nutrient content per g of processed food × g of food after processing)/(nutrient content per g of raw food × g of food before processing).

The results were submitted to analysis of variance (ANOVA) and to Tukey test for any significant differences

(P ⩽ 0.05). In all the statistical analyses, the ANOVA assumptions, such as independence and normal distribution of the residues and homogeneity of variances, were considered. The composition of carotenoids in the raw samples, in the cooked samples, and in the C. moschata ‘Menina Brasileira’ and C. maxima ‘Exposição’ pumpkin purees were determined by reverse phase HPLC ( Fig. 1). The parameters used for the identification of the peaks are shown in Table 1. As expected, epoxy-carotenoids and hydroxy-carotenoids, such as violaxanthin and lutein, were the first to elute in the reverse ABT-263 order phase column, followed by ζ-carotene, α-carotene, all-trans-β-carotene and cis-β-carotene, respectively. Peak 3 was not identified. Peaks 4 and 5 showed chromatographic data and UV–visible absorption spectra similar to those described for the carotenoids zeaxanthin and α-cryptoxanthin, respectively, as had already been noted in another study involving the same species of pumpkins ( Azevedo-Meleiro & Rodriguez-Amaya, 2007). However, because they are present in low concentrations, it was not possible to obtain isolation by OCC, therefore the spectra were not determined in other solvent systems nor were the necessary

reactions of identification carried out, and thus only one indication of the identity of those carotenoids was considered. Other minor peaks were also O-methylated flavonoid ignored. Typically, one to four carotenoids are predominant in the pumpkin species, with several other compounds detected in low concentrations or traces. The separation, identification, and quantification of these carotenoids were not the aim of this work; they can be better studied with the use of a mass spectrophotometer ( Azevedo-Meleiro & Rodriguez-Amaya, 2004). The concentration of the major carotenoids identified by HPLC in raw C. moschata ‘Menina Brasileira’ and C. maxima ‘Exposição’ pumpkins are shown in Table 2. The purity of the standard used was of 92% for lutein and 98% for α-carotene e all-trans-β-carotene, with coefficient of co-relation of (R2) of the standard curves of 0.9928, 0.9941 and 0.9933, respectively. Fig. 2 and Fig.