Screening of in vitro cytotoxicity and antioxidant potential of selected endemic plants in Turkey

Figures & data

Table 1. Mobile phase gradients in HPLC analyses and their implementation steps.

Solvent	Ratio (%)	Code
Water: Acetic acid	98: 2	A
Acetonitrile: Acetic acid	99.5: 0.5	B
Acetonitrile	100	C
90% (A), 10% (B) / 0–30 min
80% (A), 20% (B) / 30–60 min
55% (A), 45% (B) / 60–62 min
40% (B), 60% (C) / 62–77 min
90% (A), 10% (B) / 77–80 min

Table 2. Yield (%) of obtained the plant extracts.

Extract	Yield
Methanol extract of A. tokatensis	8.75
Water extract of A. tokatensis	9.50
Methanol extract of H. noeanum	10.10
Water extract of H. noeanum	10.80
Methanol extract of S. huber-morathii	9.00
Water extract of S. huber-morathii	14.40

Table 3. Quantities (μg/g) of gallic acid and protocatechuic acid phenolics in the plant extracts.

		Plant extract
Phenolic compound	Detection wavelength (nm)	Methanol extract of A. tokatensis	Water extract of A. tokatensis	Methanol extract of H. noeanum	Water extract of H. noeanum	Methanol extract of S. huber-morathii	Water extract of S. huber-morathii
Gallic acid	254	802.62	1369.34	1025.87	1747.18	1657.56	2469.16
Protocatechuic acid	254	17.64	41.33	22.85	46.27	74.01	128.87

Table 4. Antioxidant compounds of the plant extracts (µg/mg).

Treatment	Total phenol (Gallic acid equivalent)	Total flavonoid (Quercetin equivalent)
Methanol extract of A. tokatensis	34.76 ± 0.53 f	16.53 ± 0.74 f
Water extract of A. tokatensis	55.72 ± 0.57 e	38.27 ± 1.05 e
Methanol extract of H. noeanum	84.75 ± 2.30 d	49.01 ± 0.58 c
Water extract of H. noeanum	99.65 ± 1.36 c	83.39 ± 0.99 a
Methanol extract of S. huber-morathii	105.85 ± 2.13 b	71.11 ± 1.66 b
Water extract of S. huber-morathii	116.34 ± 1.51 a	40.97 ± 1.14 d

Each value is expressed as mean ± standard deviation (n = 3). Values indicated by different letters in the same column differ from each other at the level of p < 0.05.

Figure 1. DPPH radical scavenging activities of different extracts from the plants (mean ± standard deviation, n = 3) (Values indicated by different letters differ from each other at the level of p < 0.05. In the Duncan test, the highest-ranked mean is assigned the letter ‘a’, and subsequent means that are significantly different from it are assigned the next letter in the alphabet, such as ‘b’, ‘c’, and so on. Means that are not significantly different from each other are assigned the same letter, while means that are significantly different are assigned different letters). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Table 5. IC₅₀ values (mg/L) resulting from DPPH scavenging activities of the plant extracts.

Treatment	IC50 (Limits)	Slope ± Standard error of the mean (Limits)
Methanol extract of A. tokatensis	99.75 (90.17–110.80)	1.48 ± 0.06 (1.34–1.61)
Water extract of A. tokatensis	58.97 (52.37–66.21)	1.23 ± 0.06 (1.10–1.36)
Methanol extract of H. noeanum	147.75 (133.12–165.31)	1.54 ± 0.07 (1.40–1.68)
Water extract of H. noeanum	33.13 (29.51–36.87)	1.50 ± 0.07 (1.36–1.64)
Methanol extract of S. huber-morathii	48.93 (44.72–53.38)	1.79 ± 0.07 (1.64–1.94)
Water extract of S. huber-morathii	39.81 (36.00–43.79)	1.65 ± 0.07 (1.51–1.80)

Figure 2. (a) Heatmap based on IC₅₀ values for DPPH scavenging activities of plant extracts and (b) dendrogram (High and low activities were represented by red and green colour, respectively). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Figure 3. Metal chelating activities of different extracts from the plants (mean ± standard deviation, n = 3) (Values indicated by different letters differ from each other at the level of p < 0.05. In the Duncan test, the highest-ranked mean is assigned the letter ‘a’, and subsequent means that are significantly different from it are assigned the next letter in the alphabet, such as ‘b’, ‘c’, and so on. Means that are not significantly different from each other are assigned the same letter, while means that are significantly different are assigned different letters). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Table 6. IC₅₀ values (mg/L) resulting from metal chelating activities of the plant extracts.

Treatment	IC50 (Limits)	Slope ± Standard error of the mean (Limits)
Methanol extract of A. tokatensis	134.09 (115.65–158.02)	1.00 ± 0.06 (0.88–1.13)
Water extract of A. tokatensis	53.39 (43.82–64.13)	0.75 ± 0.06 (0.63–0.86)
Methanol extract of H. noeanum	240.76 (199.66–301.17)	0.96 ± 0.06 (0.83–1.09)
Water extract of H. noeanum	49.10 (42.13–56.65)	0.99 ± 0.06 (0.87–1.11)
Methanol extract of S. huber-morathii	70.22 (62.11–79.34)	1.16 ± 0.06 (1.04–1.29)
Water extract of S. huber-morathii	49.00 (42.45–56.07)	1.06 ± 0.06 (0.93–1.18)

Figure 4. (a) Heatmap based on IC₅₀ values for metal chelating activities of plant extracts and (b) dendrogram (High and low activities were represented by red and green colour, respectively). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Figure 5. Viability rates obtained by XTT analysis in MCF-7 cells treated with different extracts from the plants (mean ± standard deviation, n = 3) (Values indicated by different letters differ from each other at the level of p < 0.05. In the Duncan test, the highest-ranked mean is assigned the letter ‘a’, and subsequent means that are significantly different from it are assigned the next letter in the alphabet, such as ‘b’, ‘c’, and so on. Means that are not significantly different from each other are assigned the same letter, while means that are significantly different are assigned different letters). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Table 7. IC₅₀ values (mg/L) resulting from cytotoxic activities of the plant extracts on MCF-7 cells.

Treatment	IC50 (Limits)	Slope ± Standard error of the mean (Limits)
Methanol extract of A. tokatensis	239.33 (204.97–286.47)	1.17 ± 0.07 (1.04–1.31)
Water extract of A. tokatensis	206.94 (181.44–240.03)	1.32 ± 0.07 (1.18–1.47)
Methanol extract of H. noeanum	163.25 (144.34–186.97)	1.30 ± 0.06 (1.16–1.44)
Water extract of H. noeanum	96.94 (86.68–108.88)	1.29 ± 0.06 (1.16–1.43)
Methanol extract of S. huber-morathii	159.20 (141.89–180.53)	1.39 ± 0.07 (1.25–1.53)
Water extract of S. huber-morathii	104.99 (94.40–117.34)	1.39 ± 0.06 (1.25–1.52)

Figure 6. (a) Heatmap based on IC₅₀ values for cytotoxic activities of plant extracts and (b) dendrogram (High and low activities were represented by red and green colour, respectively). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.

Figure 7. Heatmap of percent DPPH, metal chelating and cytotoxic activities of the plant extracts. (High and low activities were represented by red and green colour, respectively). ATME: Methanol extract of A. tokatensis; ATWE: Water extract of A. tokatensis; HNME: Methanol extract of H. noeanum; HNWE: Water extract of H. noeanum; SHME: Methanol extract of S. huber-morathii; SHWE: Water extract of S. huber-morathii.