## 3 Results
### Hempeseed Screenings Bioprocessing
#### 3.2.1 Hempseed Screenings Bioprocessing Treatments Increased the Solubility of Dietary Fibre (NSP)
Within the first bioprocess: EM_1 = enzymes mixture I, BY_1= Baker's Yeast, EM + BY_1 = Baker's yeast fermentation together with enzymes mixture I, within the second bioprocess: EM_2 = enzymes mixture I, EC_2 = enzymes mixture I with cellulase, BY_2 = Baker's Yeast, EC + BY_2 = Baker's yeast fermentation together with mixture I and cellulose. Data are represented with the mean \(\pm\) S.D. Asterisks indicate the significance between the raw and bioprocessed hemspeed screenings, using One-Way ANOVA test: (*) \(p<0.05\) and (**) \(p<0.01\). The PCA (Principal Component Analysis; Figure S1) diagram of all the plant metabolites measured by targeted LC-MS/MS shows a clear discrimination between raw, first bioprocessed, and second bioprocessing treatment of hempseed screenings. The ten most abundant individual PAD with a total content of more than 10 mg/Kg is presented in Figure 2b, with benzoic acid, salicylic acid, \(p\)-hydroxybenzoic acid, vanillic acid, benzoic acid, and protocatechuic acid, benzaldehyde, protocatechuialdehyde, vanillin, and p-hydroxybenzaldehyde, and cinnamic acids, \(p\)-coumaric acid and ferulic acid. Both the first and second bioprocessing significantly increased the release in free fraction of salicylic acid, \(p\)-hydroxybenzoic acid, vanillic acid, and ferulic acid. For salicylic acid and \(p\)-hydroxybenzoic acid, a significant decrease in the alkaline-labile forms was observed after the first bioprocessing. The second bioprocessing increased the release of vanillic acid, vanillin, and \(p\)-coumaric acid in alkaline-labile form. The homoprotocatechuic acid (3, 4-dihydroxyphenylacetic acid), phloretic acid (3, 4-hydroxyphenylpropionic acid), and caffe acid (Figure 2c) were undetectable or found in trace amounts in the raw screening samples and the first bioprocessing, while they were released after the second bioprocessing, being abundant in free and alkaline-labile forms. Luteolin and catechin were the two most abundant flavonoids in the raw hempseed screenings (Figure 2b). Both the first and second bioprocessing treatments significantly increased the release of luteolin in the free form, while the release of catechin was significantly increased in the alkaline-labile form by the second bioprocessing treatment. Syringaresinol was the most abundant lignan in the raw hempseed screenings, exclusively found in the acid-labile form (72.55 \(\pm\) 16.63 mg/Kg), its release being unaffected by the bioprocessing treatments. In general, the bioprocessing treatments had a relatively limited impact on the release of the acid-labile plant metabolites, and that significantly affected the amount of plant metabolites released in the free or alkaline forms. 2.3 Hemp Screening Bioprocessing Treatments Improved the Upper Gastrointestinal Release of Several Plant Metabolites in the In Vitro Digestion Model  
Based on the results presented, the EM_1 and EC_2 bioprocessing treatments were selected to explore further their effect on in vitro digestion (IVDG), early GI bioavailability and fermentability of the hempseed screenings. The IVDG procedure involved oral, gastric, and intestinal steps, along with a dialysis process simulating the mechanical aspects of uptake. Table 3 shows the composition of the metabolites (measured by targeted LC-MS/MS) in the digestible fraction resulting from in vitro digestion of the raw and bioprocessed hemp seed screening material. Salicylic acid, p-hydroxybenzoic acid, vanillic acid, and ferulic acid are extractable mainly in the free forms, also their in vitro availability was high and was effectively improved by the bioprocessing treatments. For benzoic acid and p-hydroxybenzaldehyde, the bioprocessing treatments showed relatively limited improvement regarding their release in the free forms, this being as well as the case for their in vitro early GI bioavailability (presence in the digesta fraction).