Table 1.
Effects on total phenolics content (TPC), DPPH• scavenging activity (DPPH) and individual phenolic acid content for the different protocols for the extractable and bound phenolics of the germinated spelt seeds.
| Fraction | No. | Extraction | Extractable Fraction | Bound Fraction | In Vitro Assays (mg TE/g DW) | Phenolic Acid Content (µg/g DW) | |||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Sample–to–Solvent Ratio (w/v) | Reconstitution | SPE (mL) | Hydrolysis: 2 M NaOH (mL) | Neutralisation | SPE (mL) | TPC | DPPH | p-Coumaric Acid | trans-Ferulic Acid | Caffeic Acid Derivatives | p-Hydroxy-Benzoic Acid | ||
| Crude | #1 | MeOH 1:9 | - | - | - | - | - | 9.19 | 0.94 | 5.79 | 9.12 | 9.63 | 55.12 |
| Extractable | #2 | MeOH 1:9 | 5 mL H2O (pH 7) | 3 | na | na | na | 4.10 b | 0.52 b | 3.37 b | 6.19 b | 4.68 a | 18.59 b |
| #3 | H2O 1:9 | 5 mL H2O (pH 7) | 3 | na | na | na | 3.95 a | 0.45 a | 3.06 a | 5.43 a | 5.72 b | 14.12 a | |
| #4 | MeOH 1:6 | 5 mL H2O (pH 7) | 3 | na | na | na | 3.01 a | 0.46 a | 3.29 a | 6.06 a | 4.70 a | 16.78 a | |
| #5 | MeOH 1:9 | 30 mL 10% MeOH | 30 | na | na | na | 3.51 a | 0.56 c | 1.97 a | 4.48 a | 4.01 a | 12.51 a | |
| #6 | 5 mL H2O (pH 7) | 3 | na | na | na | 4.10 b | 0.52 a | 3.37 b | 6.19 b | 4.68 b | 18.59 b | ||
| #7 | 5 mL acidified H2O (pH 3, HCOOH) | 3 | na | na | na | 4.39 c | 0.53 b | 3.42 b | 5.92 b | 4.72 b | 23.62 c | ||
| #8 | 5 mL acidified H2O (pH 2, HCl) | 3 | na | na | na | 4.47 c | 0.55 c | 3.52 b | 6.04 b | 4.79 b | 23.93 c | ||
| #9 | 5 mL H2O (pH 7) | 3 | na | na | na | 4.10 a | 0.52 a | 3.37 a | 6.19 a | 4.68 a | 18.59 a | ||
| #10 | 5 mL H2O (pH 7) | 5 | na | na | na | 4.28 b | 0.54 b | 3.64 b | 6.97 b | 5.10 b | 20.59 b | ||
| Bound | #11 | MeOH 1:9 | na | na | 10 | HCOOH (pH 3) | 3 | 6.18 a | 0.64 a | 55.19 a | 314.99 a | 7.06 a | 27.40 a |
| #12 | na | na | 15 | HCOOH (pH 3) | 3 | 7.57 b | 0.81 c | 64.94 b | 396.05 b | 13.04 b | 36.41 b | ||
| #13 | na | na | 20 | HCOOH (pH 3) | 3 | 7.58 b | 0.74 b | 77.33 c | 534.37 c | 21.82 c | 44.16 c | ||
| #14 | na | na | 20 | HCl (pH 2) | 3 | 7.88 a | 0.73 a | 61.96 a | 363.36 a | 16.86 a | 27.68 a | ||
| #15 | na | na | 20 | HCOOH (pH 3) | 3 | 10.20 b | 1.21 b | 77.33 b | 534.37 b | 21.82 b | 44.16 b | ||
| #16 | na | na | 20 | HCOOH (pH 3) | 3 | 7.58 a | 0.74 a | 77.33 a | 534.37 a | 21.82 b | 44.16 a | ||
| #17 | na | na | 20 | HCOOH (pH 3) | 5 | 7.49 a | 0.80 b | 86.38 b | 552.13 b | 18.00 a | 42.51 a | ||
| #18 | na | na | 20 | HCOOH (pH 3) | 8 | 13.38 b | 1.17 c | 92.92 c | 563.85 c | 18.86 a | 48.96 b | ||
Data are means ± SD (n = 3). Grouping of datasets indicate data included in the statistical analysis across each dataset. No., Numbered methodology for cross-reference with Figure 1; Figure 2; na, not applicable. SPE, loading sample on SPE tube. Different small letters within a column (TPC, DPPH, phenolic acid) indicate significant differences within the individual datasets for comparisons within the extractable and within the bound contents (p < 0.05; Duncan’s multiple range tests). Note: For clarity for the statistical analysis, the following samples are repeated: #2, #6, #9; #13, #15, #16.