sampling was performed on a weekly basis during the periods September–October
2013 (n ¼ 10 samples) during the high production season. Sampling then continued
during the period April–October 2014 (n ¼ 17 samples) and February–October 2015
(n ¼ 17 samples).
18.2.3 NSAIDs Removal Evaluation
For the evaluation of NSAIDs removal in the constructed wetland, an in vitro
analytical method was developed and an analysis of these antiphlogistics in a real
system was performed thereafter. Metabolic potential of cell cultures of Melilotus
officinalis and Rheum palmatum in vitro was verified in Phragmites australis cells,
tissues, and whole plants, and finally tested in the CW. Some measurements of
NSAIDs concentrations in this CW were reported by Marsik et al. (2015).
The sampling and preparation procedure of the samples was done according to
Marsik et al. (2015). The samples were collected in the period June–September 2014
and February–June 2015 (15 samples in total) from the same sampling points as
mentioned above. Wastewater samples were analyzed using UPLC-MS/MS. For the
analysis, the samples were first acidified with acetic acid at pH 2.5 and then they
were filtered through a nylon membrane filter (0.45 μm). For quantitative analysis,
system Q-Trap 4000 (AB Sciex, USA) was used. The sample (25 mL) was applied
on solid phase extraction columns (Strata C8 (55 μm, 70 A), 500 mg/3 mL,
Phenomenex, USA) preconditioned with 5 mL of methanol and 10 mL of deionized
water. The columns were then washed with acidified water (acetic acid, pH 2) and
eluted with 5 mL of methanol. The samples were then evaporated under stream of
nitrogen to dryness and stored at À80
C. Before the analysis, the samples were
dissolved in 1 mL of methanol and then dissolved 100Â for analysis.
18.3 Results and Discussion
18.3.1 Performance of the Hybrid Constructed Wetland
In 2013 the monitoring was focused mainly on the period of wine harvesting and
processing. In Table 18.2, mean concentrations of COD, BOD 5 , and TSS are
summarized. The results show that the discharge limits (see in Table 18.3) were
fulfilled at the outflow from the CW filters; however, the quality of the water
decreased in the stabilization ponds when the COD increased from 40 mg L
À1 at
the outlet from HF2 to 181 mg L
À1 at the outlet from SP3. As for the TSS, the C
out
increased from average 3 mg L
À1 at the outlet from HF2 to 1047 mg L
À1 at the outlet
from SP3. This increase was caused mainly by the growth of algae and decay of
plants in the stabilization ponds (see Fig. 18.2). Shepherd et al. (2001) reported very
400
M. Šereš et al.
2013 (n ¼ 10 samples) during the high production season. Sampling then continued
during the period April–October 2014 (n ¼ 17 samples) and February–October 2015
(n ¼ 17 samples).
18.2.3 NSAIDs Removal Evaluation
For the evaluation of NSAIDs removal in the constructed wetland, an in vitro
analytical method was developed and an analysis of these antiphlogistics in a real
system was performed thereafter. Metabolic potential of cell cultures of Melilotus
officinalis and Rheum palmatum in vitro was verified in Phragmites australis cells,
tissues, and whole plants, and finally tested in the CW. Some measurements of
NSAIDs concentrations in this CW were reported by Marsik et al. (2015).
The sampling and preparation procedure of the samples was done according to
Marsik et al. (2015). The samples were collected in the period June–September 2014
and February–June 2015 (15 samples in total) from the same sampling points as
mentioned above. Wastewater samples were analyzed using UPLC-MS/MS. For the
analysis, the samples were first acidified with acetic acid at pH 2.5 and then they
were filtered through a nylon membrane filter (0.45 μm). For quantitative analysis,
system Q-Trap 4000 (AB Sciex, USA) was used. The sample (25 mL) was applied
on solid phase extraction columns (Strata C8 (55 μm, 70 A), 500 mg/3 mL,
Phenomenex, USA) preconditioned with 5 mL of methanol and 10 mL of deionized
water. The columns were then washed with acidified water (acetic acid, pH 2) and
eluted with 5 mL of methanol. The samples were then evaporated under stream of
nitrogen to dryness and stored at À80
C. Before the analysis, the samples were
dissolved in 1 mL of methanol and then dissolved 100Â for analysis.
18.3 Results and Discussion
18.3.1 Performance of the Hybrid Constructed Wetland
In 2013 the monitoring was focused mainly on the period of wine harvesting and
processing. In Table 18.2, mean concentrations of COD, BOD 5 , and TSS are
summarized. The results show that the discharge limits (see in Table 18.3) were
fulfilled at the outflow from the CW filters; however, the quality of the water
decreased in the stabilization ponds when the COD increased from 40 mg L
À1 at
the outlet from HF2 to 181 mg L
À1 at the outlet from SP3. As for the TSS, the C
out
increased from average 3 mg L
À1 at the outlet from HF2 to 1047 mg L
À1 at the outlet
from SP3. This increase was caused mainly by the growth of algae and decay of
plants in the stabilization ponds (see Fig. 18.2). Shepherd et al. (2001) reported very
400
M. Šereš et al.