Univariate analysis of variance was used for all data in determining significant differences. For descriptive analysis data, multivariate analysis of variance was used prior to ANOVA to determine the main treatment effect. ANOVA was used for judge, treatment, and replicate effects along with a pseudo mixed model. Fisher’s least significant difference was used for pairwise comparisons of means. Statistical significance was set at 5% for all tests.Analysis of Brix, pH, and TA of the musts showed minimal differences among treatments for each variety . There were no significant differences for all three parameters of the BA must and only the reject treatment for GN had a significantly higher TA; however, this difference was not large. It is possible that this difference could be the result of the inclusion of underripe berries in the must, which have a higher TA. Raisins were also rejected from the sorter, which are high in sugar and could have compensated for the difference in sugar from the less ripe berries. The CS must exhibited the most differences among treatments, which was unexpected considering this variety had the lowest percentage of rejected fruit . The Brix was significantly higher in the sorted treatment compared to the control and reject treatments. This may indicate that the sorter was effective at removing less ripe berries for CS. The pH also differed significantly among treatments for CS; pH was highest in the reject must at 3.8, followed by sort and control at 3.71 and 3.67 respectively. Although the difference in pH between the sort and control was statistically significant, they are very similar with only a 0.04 pH unit difference. Overall, growing blueberries in pots the differences seen in the must chemistry were minimal and likely made little to no difference in the progression of the wines.
It is possible that the reject must composition was made to be more similar to the control and sort treatments due to the addition of juice that accumulated in the vibrating table trays. If this was not done perhaps there would be more differences in must composition when comparing the reject to the sort and control treatments. Wine chemical compositions are shown in Table 5. All wines progressed consistently through fermentation and fermented dry with less than 1 g/L residual sugar. For the most part, wine chemical compositions are similar among treatments for each variety, especially between the sort and control treatments. However, there are some important exceptions. This mostly corresponds to differences in the starting sugar content, although there is a discrepancy as BA reject wines were not significantly lower in Brix. However, Brix was determined after mixing of the must, and especially if a significant number of raisins were present, soak up in the next 24 h could have resulted in sugar increases. The malolactic fermentations for GN and CS wines progressed to completion; however, the control and sort treatments for BA did not finish and were left with close to 1 g/L malic acid for each of the treatments . This is likely due to the high ethanol content in addition to high TA in the wines which can inhibit malolactic bacteria. This would also explain why the reject treatment for BA progressed further in the malolactic fermentation given that these wines were lower in ethanol content and TA. This difference could have important implications for the sensory analysis of BA wines. The volatile acidity for the CS reject treatment was significantly higher than that for the control and sort treatments . The sensory threshold has been reported to be approximately 0.8 g/L for red table wines, therefore, this discrepancy may not have a large impact on sensory analysis.
It was surprising that GN musts/wines showed few significant differences despite having the largest rate of rejection . It is possible that sorting parameters were too aggressive when processing GN, which may have inadvertently led to the rejection of optimal fruit. As previously mentioned, there was significant variation in color for GN fruit . It may also be possible that observed color difference in GN fruit did not correlate well with sugar content. This would mean that optical sorting based on color for GN fruit from this vineyard is potentially less effective than for the other varieties.Differences among treatments were observed in total phenolics, tannin, and anthocyanin content as measured by the Adams-Harbertson assay . In general, the reject wines were higher than control and sort wines in total phenolics and tannin, and lower in anthocyanin. This may be explained by the inclusion of MOG in the reject fermentations which can lead to greater extraction of phenolics. Lower anthocyanin levels wereobserved in the reject wines for all varieties. This is most likely due to the inclusion of green, underripe berries, which contain less anthocyanin.In general, the results from the RP-HPLC analysis of phenolics agree with the results obtained from the Adams-Harbertson assay . Higher levels of most phenolic compounds were observed in the reject treatments. Concentrations of gallic acid and catechin were higher in the reject treatments for all three varieties and dimer B1 was higher in reject wines for BA and CS. Less ripe berries have been shown to contain more of these compounds, which can explain this trend. Higher levels of identified flavan- 3-ols were also observed in the reject treatments of BA and CS wines, which is also in agreement with results found by Obreque-Slier. An interesting trend was found in relation to the proportions of simple hydroxycinnamic acids and their respective tartaric acid esters. All the reject treatments had very low amounts of caftaric and coutaric acid compared to caffeic and p-coumaric acid. It is possible that hydroxycinnamoyl esterase, the enzyme responsible for hydrolyzing the ester linkage, had a greater activity in the reject wines, possibly due to differences in pH . Another possibility is that there could be higher levels of this enzyme in less-ripe fruit. The reject wines for all three varieties were also significantly lower in anthocyanin, which matches results obtained by the Adam-Harbertson assay.
Although reject wines had higher levels of most phenolic compounds, this did not lead to large differences between sorted and control wines. It is likely that not enough material was removed during processing for there to be a significant effect. This may also explain why there were no significant differences in anthocyanin content between sorted and control wines despite reject wines being significantly lower. Perhaps a greater effect would be observed with more aggressive sorting parameters and/or fruit with more variability. Overall, the levels of most phenolic compounds identified were very similar between the sort and control treatments. It can be concluded that optical sorting had little impact on the composition of phenolic compounds between sorted and control wines in all three varieties tested.For CS wines, 37 volatile compounds were identified, 20 of which differed significantly among treatments ; however, only one compound differed significantly between wines made from sorted and control treatments . A Principle Component Analysis biplot plot of significant compounds is presented in Figure 1. It appears the separation is driven primarily by ethyl esters on the left and higher alcohols on the right. Most ethyl esters have higher concentrations in wines made from control and sort treatments . Esters in wine can be formed by an acid catalyzed esterification reaction between an acid and alcohol.Higher amounts of either acid or alcohol can result in increased formation of esters. Wines made from control and sorted treatments had higher ethanol content than wines made from the reject treatment, which would explain this trend. Another important trend is the association of reject treatment wines with a larger concentration of higher alcohols. The suspended solids concentration was significantly higher in reject treatment musts which may explain the difference in the concentration of higher alcohols among the treatments, as previous research has shown that suspended solids during fermentation can lead to greater production of higher alcohols. PCA loading and score plots of volatile analysis for BA wines are given in Figure 2. Thirty-seven compounds were identified, drainage gutter and nine differed significantly among treatments . Again, separation seems to be driven by the proportionally larger presence of higher alcohols in the reject treatments. Like the CS reject musts, the BA reject musts also had significantly higher levels of soluble solids compared to the sort and control treatments . Although most ethyl esters did not differ significantly among treatments for BA, there was a general trend indicating that ethyl ester content was higher in the control and sort treatments . A PCA biplot using all identified ethyl esters and higher alcohols is provided in Figure S1 and there is a clear trend in the separation of these compounds. This agrees with the previous discussion regarding ethyl ester content in the CS wines. The BA control and sort treatments had significantly higher ethanol content compared to the rejects so it is expected that ethyl ester concentration would be higher as well. One exception to this trend is that ethyl lactate was significantly higher in the reject treatment.
The reject wines completed ML fermentation, but the control and sort wines got stuck with almost 1 g/L malic acid . Therefore, ethyl lactate is significantly higher in reject wines because there was more lactic acid present from the conversion of nearly all the malic acid.For GN wines, 32 compounds were identified, nine of which differed significantly among treatments and four differed significantly between sort and control treatments. The same trend was observed for higher alcohols for GN as for the other varieties driving separation in the PCA plot . The concentrations of cis-3-hexen-1-ol, trans-3- hexen-1-ol, and hexanol were all significantly higher in the reject treatments. Again, this is most likely due to higher suspended solids content in the reject treatment musts . The trend with ethyl esters was not observed for GN wines, most likely because all treatments had similar ethanol content . Overall, the results indicate that optical sorting had a minimal effect on the aroma profile for all three varieties, particularly when comparing sort and control treatments.Given the uniformity of chemical results among biological replications, it is fair to assume that the two replications used for descriptive analysis are representative, and the chemical results can therefore be used to discuss sensory trends. MANOVA was performed and revealed a non-significant treatment effect for all three varieties . From this result, it can be concluded that all three treatments for each variety were similar in sensory properties. Despite this result, ANOVA was carried out on individual attributes and some significant differences were found for each variety . For GN, only one attribute out of twenty differed significantly among treatments. It is possible that sensory analysis was done too soon after the wines were bottled and the levels of molecular sulfur dioxide may have been above sensory threshold of about 2 mg/L. Figure 4 gives a PCA biplot with all attributes from the GN descriptive analysis panel. There are no clear trends from the PCA; therefore, it can be concluded from MANOVA, ANOVA, and PCA results that all treatments lead to wines of similar character for GN wines. When ANOVA was performed on data from the BA descriptive analysis panel, three out of twenty-six attributes were found to be significantly different among treatments. “Alcohol hotness” had a significant judge-by-treatment interaction. Results from the pseudo mixed model indicated the interaction effect was more important than the treatment effect. Thus, “alcohol hotness” will not be included in any further discussion of significant attributes for BA wines. The significant difference in malic acid content in the wines among treatments appears to have had little impact on sensory evaluation given that there was no significant difference in the perception of sourness in the wines. From the PCA generated from BA descriptive analysis results , the control and sort wines appear to be correlated more closely with “alcohol” . Wines made from these treatments were higher in ethanol content, which may explain this trend. However, the small number of significant attributes indicate that BA wines made by different treatments were very similar in sensory properties.For the CS descriptive analysis panel, three out of twenty-two attributes were found to be significantly different when ANOVA was performed. A PCA biplot from the CS panel is provided in Figure 6.