Phenolic Composition, Essential Oil, and Antioxidant Activity in the Aerial Part of Artemisia Herba-Alba from Several Provenances: A Comparative Study

Figures & data

TABLE 1 Geographical characteristics of the four sites of collect of A. herba-alba

	Longitude	Latitude	Elevation (m)	Bioclimatic zone
Kef	8°43’14.07’’E	36°10’18.66’’N	529	Semi-arid superior
Boukornine	10°22’54.27’’E	36°42’25.33’’N	70	Semi-arid superior
Kairouan	10°29’2.23’’E	35°44’21.88’’N	122	Semi-arid inferior
Kasserine	8°50’20.55’’E	35°10’41.17’’N	2211	Arid superior

TABLE 2 Essential oil yields and total polyphenols and flavonoids contents from four A. herba-alba accessions

	Accessions
	Kef	Boukornine	Kairouan	Kasserine
Essential oil yield (%, w/w)	1.11 ± 0.07b	1.15 ± 0.07a	0.35 ± 0.06d	0.77 ± 0.01c
Total polyphenols (mg GAE/g DW)	8.38 ± 0.75d	14.03 ± 1.33c	16.80 ± 1.86b	17.93 ± 1.47a
Total flavonoids (mg CE/g DW)	16.15 ± 1.44d	32.58 ± 4.21c	43.59 ± 3.96a	39.77 ± 4.18b

The results represent the average of three replicates (n = 3);

Letters (a–d) in the same line indicate significant differences at p < 0.05;

DW: dry weight basis.

TABLE 3 Analysis of variance analysis and phenolic composition (expressed in%, w/w, and in µg/g dry weight) of methanolic extracts from four A. herba-alba accessions

Compounds	Accessions
	Kef		Boukornine		Kairouan		Kasserine
	µg/g DW	%	µg/g DW	%	µg/g DW	%	µg/g DW	%
Tannic acid	0.38 ± 0.03^B	4.06 ± 0.35^b	0.18 ± 0.01^D	1.01 ± 0.09^d	0.30 ± 0.02^C	2.21 ± 0.17^c	0.87 ± 0.07^A	5.50 ± 0.41^a
Gallic acid	0.39 ± 0.04^C	4.20 ± 0.38^a	0.44 ± 0.04^B	2.48 ± 0.18^c	0.32 ± 0.02^D	2.36 ± 0.20^d	0.62 ± 0.04^A	3.94 ± 0.41^b
Chlorogenic acid	0.31 ± 0.02^D	3.36 ± 0.37^c	0.67 ± 0.05^B	3.76 ± 0.42^b	0.37 ± 0.03^C	2.75 ± 0.30^d	1.45 ± 0.12^A	9.13 ± 1.01^a
Caffeic acid	0.58 ± 0.06^C	6.28 ± 0.72^c	1.17 ± 0.10^B	6.56 ± 0.52^b	0.18 ± 0.13^D	1.32 ± 0.12^d	2.22 ± 0.21^A	14.04 ± 1.22^a
Vanilic acid	nd	nd	nd	nd	0.31 ± 0.02	2.27 ± 0.21	nd	nd
Syringic acid	0.59 ± 0.06^B	6.33 ± 0.71^b	0.56 ± 0.05^C	3.13 ± 0.28^c	nd	nd	1.36 ± 0.12^A	8.56 ± 0.77^a
Ferrulic acid	0.52 ± 0.05^B	5.56 ± 0.43^a	0.47 ± 0.04^C	2.64 ± 0.23^c	0.27 ± 0.03^D	2.01 ± 0.17^d	0.57 ± 0.05^A	3.63 ± 0.35^b
p-Coumaric acid	0.58 ± 0.06^D	6.19 ± 0.73^d	2.85 ± 0.23^B	16.00 ± 1.53^b	3.15 ± 0.34^A	23.34 ± 2.44^a	1.14 ± 0.12^C	7.17 ± 0.65^c
Rosmarinic acid	0.37 ± 0.03^C	3.99 ± 0.41^d	1.04 ± 0.11^A	5.87 ± 0.63^c	0.97 ± 0.09^B	7.17 ± 0.76^a	0.95 ± 0.08^B	5.97 ± 0.65^b
trans-Cinnamic acid	0.98 ± 0.08^C	10.52 ± 1.12^a	1.29 ± 0.12^A	7.27 ± 0.82^b	0.21 ± 0.02^D	1.55 ± 0.14^d	1.12 ± 0.10^B	7.06 ± 0.63^c
Catechin	0.30 ± 0.03^C	3.23 ± 0.35^c	0.67 ± 0.06^A	3.78 ± 0.38^b	0.52 ± 0.04^B	3.84 ± 0.41^a	0.28 ± 0.02^D	1.76 ± 0.14^d
Epicatechin	0.26 ± 0.02^C	2.77 ± 0.24^b	0.93 ± 0.08^A	5.26 ± 0.52^a	0.71 ± 0.06^B	5.28 ± 0.49^a	nd	nd
Naringenin	0.31 ± 0.02^D	3.36 ± °.28^d	2.40 ± 0.23^B	13.51 ± 1.42^b	2.73 ± 0.25^A	20.19 ± 1.89^a	0.77 ± 0.07^C	4.83 ± 0.42^c
Myristin	0.41 ± 0.03^C	4.45 ± 0.41^c	0.60 ± 0.05^B	3.39 ± 0.34^d	0.61 ± 0.05^B	4.51 ± 0.36^b	0.99 ± 0.09^A	6.25 ± 0.53^a
Quercetin	0.20 ± 0.01^B	2.11 ± 0.18^b	0.19 ± 0.02^B	1.05 ± 0.11^d	0.15 ± 0.01^C	1.10 ± 0.10^c	0.58 ± 0.06^A	3.68 ± 2.89^a
Kaempherol	1.08 ± 0.10^A	11.58 ± 1.12^a	0.23 ± 0.03^C	1.29 ± 0.11^c	0.10 ± 0.01^D	0.75 ± 0.06^d	0.54 ± 0.05^B	3.38 ± 0.29^b
Apigenin	0.62 ± 0.05^D	6.61 ± 0.57^b	1.86 ± 0.16^A	10.47 ± 1.21^a	0.83 ± 0.07^B	6.15 ± 0.57^c	0.65 ± 0.06^C	4.08 ± 0.39^d
Classes
Phenolic acids	4.70 ± 0.39^D	50.50 ± 5.24^b	8.66 ± 0.78^B	48.72 ± 4.37^c	6.07 ± 0.62^C	44.98 ± 4.73^d	10.30 ± 1.02^A	65.00 ± 6.48^a
Flavan-3-ols	0.56 ± 0.04^C	6.01 ± 0.53^c	1.61 ± 0.13^A	9.03 ± 0.86^b	1.23 ± 0.11^B	9.12 ± 0.87^a	0.28 ± 0.03^D	1.76 ± 0.16^d
Flavanones	0.31 ± 0.03^D	3.36 ± 0.31^d	2.40 ± 0.24^B	13.51 ± 1.22^b	2.73 ± 0.25^A	20.19 ± 1.87^a	0.77 ± 0.07^C	4.83 ± 0.38^c
Flavonols	1.69 ± 0.14^B	18.14 ± 1.65^a	1.02 ± 0.09^C	5.72 ± 0.45^d	0.86 ± 0.07^D	6.36 ± 0.58^c	2.11 ± 0.22^A	13.31 ± 1.21^b
Flavones	0.62 ± 0.06^D	6.61 ± 0.57^b	1.86 ± 0.15^A	10.47 ± 0.95^a	0.83 ± 0.08^B	6.15 ± 0.57^c	0.65 ± 0.05^C	4.08 ± 0.38^d

Quantities and percentages of phenolic compounds represent the average of three replicates (n = 3);

nd: not detected;

Capital letters (A–D) and small letters (a–d) in the same line indicate significant differences at p < 0.05;

DW: dry weight basis.

TABLE 4 Calibration curves, correlation coefficients (R²), detection limits, and quantification limits for high-performance liquid chromatography analysis for major phenolics identified from A. herba-alba

Compounds	Equation for regression line	Correlation coefficient (R^2)	Limit of detection (LOD) (µg/mL)	Limit of quantitation (LOQ) (µg/mL)
p-coumaric acid	y = 3.378x + 39.456	0.992	4.994	16.648
Caffeic acid	y = 7.863x + 196.8	0.992	6.109	20.363
trans-Cinnamic acid	y = 29.97x – 70.06	0.999	0.854	2.849
kaempferol	y = 20.01x – 62.93	0.994	2.353	7.843
Apigenine	y = 23.46x – 259.8	0.975	6.754	22.514

FIGURE 1 Hierarchical clustering of A. herba-alba accessions based on their phenolic composition.

TABLE 5 Analysis of variance analysis and essential oils composition (%) from four A. herba-alba accessions

Compounds	RI^a	RI^b	Accessions
			Kef	Boukornine	Kairouan	Kasserine
α-Pinene	931	1017	0.69 ± 0.11^a	0.38 ± 0.06^b	0.37 ± 0.10^b	0.73 ± 0.15^a
Camphene	950	1048	1.83 ± 3.11^bc	4.66 ± 0.63^a	3.33 ± 0.75^ab	0.25 ± 0.05^c
o-Cymene	976	1236	0.42 ± 0.01^b	0.67 ± 0.04^a	0.18 ± 0.02^c	0.52 ± 0.05^ab
Sabinene	977	1112	0.20 ± 0.21^bc	0.85 ± 0.14^a	ND	0.32 ± 0.10^b
β-Pinene	980	1095	0.53 ± 0.28^ab	0.36 ± 0.03^bc	0.15 ± 0.07^c	0.67 ± 0.07^a
α-Terpinene	1018	1187	0.16 ± 0.17^bc	0.12 ± 0.02^bc	0.03 ± 0.02^c	0.26 ± 0.10^a
α-Phellandrene	1023	1169	0.21 ± 0.17^bc	0.27 ± 0.01^bc	0.12 ± 0.01^c	0.31 ± 0.05^a
p-Cymene	1026	1272	0.18 ± 0.09^b	0.13 ± 0.00^b	0.47 ± 0.07^a	0.13 ± 0.02^b
Limonene	1030	1194	7.29 ± 0.28^a	9.27 ± 0.07^a	5.88 ± 0.50^a	4.57 ± 0.43^a
1-8-Cineole	1033	1210	0.16 ± 0.05^a	0.05 ± 0.00^b	0.08 ± 0.01^b	0.19 ± 0.03^a
γ-terpinene	1062	1249	0.63 ± 0.09^b	0.67 ± 0.02^b	1.16 ± 0.12^a	0.67 ± 0.07^b
Terpinolene	1078	1314	0.12 ± 0.02^a	0.06 ± 0.01^a	0.33 ± 0.04^a	2.71 ± 0.19^a
α-Thujone	1088	1426	13.93 ± 0.63^a	11.62 ± 1.20^a	13.23 ± 0.31^a	11.62 ± 1.45^a
Linalool	1098	1557	1.61 ± 0.57^a	0.72 ± 0.23^b	1.62 ± 0.39^a	1.78 ± 0.06^a
Fenchol	1099	1405	9.04 ± 0.97^a	13.85 ± 0.68^a	7.51 ± 1.94^a	12.69 ± 1.26^a
β-thujone	1103	1439	5.44 ± 0.25^bc	2.73 ± 0.28^bc	0.16 ± 0.04^c	8.04 ± 0.71^a
p-Menth-2-en-1-ol	1108	1622	0.65 ± 0.15^a	0.55 ± 0.04^a	0.77 ± 0.56^a	0.69 ± 0.08^a
Chrysanthenone	1110	1481	3.73 ± 0.58^a	ND	6.81 ± 0.10^a	5.57 ± 0.66^a
cis-Sabinol	1115	1812	0.52 ± 0.05^a	0.56 ± 0.38^a	0.42 ± 0.04^a	0.47 ± 0.05^a
trans-Pinocarveol	1126	1693	2.32 ± 0.50^a	2.55 ± 0.56^a	1.54 ± 0.04^a	2.63 ± 0.03^a
Camphor	1143	1495	12.37 ± 1.71^ab	31.51 ± 2.03^a	24.97 ± 0.83^a	0.64 ± 0.04^b
Borneol	1165	1719	0.31 ± 0.26^a	0.09 ± 0.00^a	1.64 ± 0.06^a	0.43 ± 0.06^a
Myrtenal	1172	1658	2.79 ± 0.09^ab	0.62 ± 0.17^b	1.29 ± 0.22^b	3.94 ± 0.31^a
Terpinen-4-ol	1178	1589	1.22 ± 0.57^a	1.29 ± 0.05^a	0.99 ± 0.39^a	1.45 ± 0.17^a
cis-Piperitol	1181	1746	0.23 ± 0.03^a	0.24 ± 0.05^a	0.20 ± 0.01^a	0.25 ± 0.08^a
p-cymen-8-ol	1183	2067	0.76 ± 0.01^a	0.59 ± 0.02^a	0.09 ± 0.04^a	0.97 ± 0.05^a
Myrtenol	1187	1837	0.37 ± 0.15^a	0.07 ± 0.03^a	0.31 ± 0.26^a	0.40 ± 0.04^a
α-Terpineol	1189	1711	0.65 ± 0.54^a	0.95 ± 0.16^a	0.87 ± 0.14^a	0.86 ± 0.09^a
Piperitone	1226	1702	0.20 ± 0.06^b	0.17 ± 0.02^b	1.07 ± 0.81^a	0.20 ± 0.04^b
Carvone	1242	1759	0.97 ± 0.63^a	0.43 ± 0.02^a	0.63 ± 0.14^a	0.67 ± 0.28^a
Phellandral	1278	1732	0.15 ± 0.10	0.03 ± 0.01	0.07 ± 0.02	0.18 ± 0.03
Eugenol	1356	2186	0.79 ± 0.06^ab	1.20 ± 0.39^a	0.19 ± 0.07^b	1.01 ± 0.14^a
Z-Jasmone	1371	1931	2.30 ± 0.09^a	0.36 ± 0.03^a	0.33 ± 0.03^a	2.60 ± 0.11^a
α-Copaene	1379	1461	0.20 ± 0.04^b	0.36 ± 0.03^a	0.22 ± 0.12^b	0.18 ± 0.02^b
cis-Threo-davana furan	1419	1769	0.40 ± 0.33^a	0.28 ± 0.02^a	0.61 ± 0.01^a	0.54 ± 0.06^a
β-Caryophyllene	1421	1610	0.46 ± 0.10^b	0.68 ± 0.01^b	1.31 ± 0.09^a	0.40 ± 0.05^b
Aristolene	1423	1476	0.09 ± 0.05^a	0.16 ± 0.02^a	0.10 ± 0.02^a	0.10 ± 0.04^a
Nordavanone	1451	1645	7.04 ± 0.02^a	1.36 ± 0.37^b	1.26 ± 0.20^b	9.44 ± 0.70^a
Germacrene-D	1480	1718	3.20 ± 0.39^ab	4.56 ± 0.37^bc	5.89 ± 0.79^a	2.24 ± 0.62^c
Davana ether	1489	1883	0.97 ± 0.04^a	0.15 ± 0.02^b	0.24 ± 0.01^b	1.22 ± 0.20^a
β-Bisabolene	1503	1491	1.44 ± 0.27^a	ND	ND	1.52 ± 0.20^a
Davana ether	1507	1915	1.21 ± 0.10^a	0.07 ± 0.02^b	0.15 ± 0.04^b	1.44 ± 0.46^a
cis-Davanone	1557	2023	0.31 ± 0.22^b	0.08 ± 0.02^b	0.17 ± 0.04^b	0.67 ± 0.35^a
β-Caryophyllene epoxide	1577	1946	0.44 ± 0.42^a	0.02 ± 0.00^a	0.11 ± 0.06^a	0.46 ± 0.31^a
Viridiflorol	1592	2104	2.08 ± 1.55^a	0.12 ± 0.02^b	1.76 ± 0.54^a	2.58 ± 0.33^a
t-Muurolol	1633	2176	0.16 ± 0.03^a	0.24 ± 0.09^a	0.37 ± 0.18^a	0.20 ± 0.10^a
Monoterpene hydrocarbons			12.42 ± 1.36^b	17.50 ± 1.06^a	12.10± 1.35^c	11.33 ± 0.95^d
Oxygenated monterpenes			66.58 ± 7.75^b	70.29 ± 5.48^a	65.79 ± 4.97^c	65.54 ± 3.21^d
Sesquiterpene hydrocarbons			3.95 ± 0.17^c	5.76 ± 0.26^b	7.52 ± 0.87^a	2.92 ± 0.06^d
Oxygenated sesquiterpenes			7.02 ± 0.34^b	0.96 ± 0.06^d	3.42 ± 0.25^c	8.63 ± 0.47^a
Phenylpropanoids			0.79 ± 0.06^ab	1.20 ± 0.39^a	0.19 ± 0.07^b	1.01 ± 0.14^a

ND: not detected;

Volatile compounds percentages in the same line with different letters (a–d) are significantly different at p < 0.05;

RI^a: Order of elution in apolar column (HP-5);

RI^b: Order of elution in polar column (HP-Innowax);

Boldface: Major compounds.

FIGURE 2 Dendogram obtained by cluster analysis of essentials oils of A. alba alba from different regions based on their chemical composition.

TABLE 6 Antiradical activity IC₅₀ (µg/mL) and reducing power CE₅₀ (mg/mL) of A. herba-alba extracts

		Accessions
		Boukornine	Kef	Kairouan	Kasserine
DPPH activity	Methanol extract	3	4	3	3
	Essential oil	1300	1300	110	400
	BHT	16 ± 1.89
Reducing power	Methanol extract	0.12	0.36	0,10	0.10
	Essential oil	2	2.9	2.6	1.2
	Ascorbic acid	0.032

Szabo, M.R.; Radu, D.; Gavrilas, S.; Chambre, D.; Iditoiu, C. Antioxidant and Antimicrobial Properties of Selected Spice Extracts. International Journal of Food Properties 2010, 13, 535–545.

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