Chemical composition and pharmacological properties of the essential oils obtained seasonally from Lippia thymoides

Figures & data

Table 1. Monthly climatological data and content of essential oils from leaves of L. thymoides obtained in the four seasons.

		Essential oil code
	Information	OAA	OJW	OOS	OJS
Collect	Date of collect	04/28/2009	07/24/2009	10/28/2009	01/28/2010
	Average temperature (°C)	25.5	23.3	26.9	28.9
	Relative humidity (%)	82	84	70	72
	Rainfall (mm)	281.1	107.4	74.1	119.3
	Total insolation (h)	357.1	242.2	382.4	420.7
Content of essential oils	Plant dry mass (g)	166.1	355.0	256.0	133.4
	Residual humidity (%)	10.79	8.72	9.45	8.52
	Oil volume (mL)	4.0	8.2	7.5	2.5
	Oil content (%)	2.41	2.31	2.93	2.14

OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; and OJS, essential oil from L. thymoides collected in summer.

Table 2. Chemical composition of the essential oils from the leaves of L. thymoides.

			OAA		OJW		OOS		OJS
Formula	Constituent	KIL	KIC	%	KIC	%	KIC	%	KIC	%
C10H16	α-Thujene	930	930	0.11	930	0.09	930	0.27	–	–
C10H16	α-Pinene	932	938	1.81	938	0.94	938	2.38	931	1.42
C10H16	Camphene	946	953	5.44	953	2.64	954	5.66	943	3.25
C10H16	Sabinene	969	976	1.31	978	0.93	976	1.72	962	0.67
C10H16	β-Pinene	974	980	1.16	980	0.64	980	1.43	965	1.70
C10H16	β-Myrcene	988	991	0.94	991	0.49	991	0.94	–	–
C10H16	α-Phellandrene	1002	1007	0.08	1006	0.07	–	–	–	–
C10H16	3-Carene	1008	1013	0.27	1013	0.23	1013	0.26	–	–
C10H16	o-Cimene	1022	1026	0.47	1026	0.91	1027	2.69	1027	1.12
C10H16	Limonene	1029	1032	3.63	1032	2.42	1032	3.75	1032	1.67
C10H18O	1,8-Cineole	1031	1034	3.04	1034	3.88	1034	4.50	1034	1.86
C10H16	γ-Terpinene	1059	1061	0.51	1061	0.65	1062	1.01	1062	0.74
C10H16	Terpinolene	1088	1090	0.86	1090	0.47	1090	0.46	–	–
C10H16O	Pinocarveol	1139	–	–	1141	0.12	1141	0.22	–	–
C10H16O	Camphor	1146	1146	8.61	1146	4.92	1146	3.22	1146	4.52
C10H18O	Borneol	1169	1168	5.33	1168	4.45	1168	7.36	1168	6.46
C10H18O	4-Terpinenol	1177	1178	0.83	1178	0.59	1179	0.85	–	–
C10H18O	α-Terpineol	1188	–	–	1190	0.23	–	–	–	–
C10H14O	Verbenone	1205	–	–	1203	0.79	–	–	–	–
C12H20O2	Bornyl acetate	1288	–	–	1286	0.35	–	–	–	–
C10H14O	Thymol	1290	–	–	1290	0.12	1291	0.21	1290	0.27
C15H24	δ-Elemene	1338	–	–	1340	0.38	–	–	–	–
C15H24	α-Cubebene	1348	1352	0.63	1352	0.60	1353	0.77	1353	0.65
C15H24	Copaene	1376	1378	2.47	1378	2.55	1379	3.38	1379	2.42
C15H24	β-Bourbonene	1388	–	–	1386	0.28	1387	0.40	–	–
C15H24	β-Cubebene	1388	–	–	–	–	1392	0.28	–	–
C15H24	β-Elemene	1390	–	–	1392	0.24	–	–	–	–
C15H24	α-Bergamotene	1412	1406	0.54	1407	0.29	1407	0.40	–	–
C15H24	α-Gurjunene	1409	–	–	1411	0.34	1412	0.38	1412	0.82
C15H24	α-Cedrene	1411	1411	0.30	1414	0.52	1415	0.82	–	–
C15H24	β-Caryophyllene	1419	1422	17.22	1423	26.27	1423	21.20	1423	19.28
C15H24	α-Caryophyllene	1454	1457	5.05	1456	3.99	1457	3.06	1457	5.48
C15H24	β-Farnesene	1456	–	–	1463	0.44	–	–	–	–
C15H24	Alloaromaden-drene	1460	–	–	1469	0.04	–	–	1465	0.57
C15H24	Germacrene D	1485	1483	4.80	1483	6.18	1484	4.72	1484	5.39
C15H24	α-Zingiberene	1493	–	–	1493	1.43	–	–	–	–
C15H24	α-Muurolene	1500	1506	0.57	1498	1.67	1507	1.06	1509	2.00
C15H22	Cuparene	1504	–	–	1506	0.78	1510	0.88	–	–
C15H24	β-Bisabolene	1505	–	–	1509	0.89	–	–	–	-
C15H24	Z-α-Bisabolene	1507	1509	1.12	–	–	–	–	–	–
C15H22	Calamenene	1529	1525	4.30	1525	3.53	1526	4.75	1526	3.83
C15H24	Germacrene B	1561	–	–	–	–	1561	0.88	1561	1.04
C15H26O	trans-Nerolidol	1561	1564	1.57	1564	3.59	1565	1.52	1565	1.69
C15H24O	Spathulenol	1578	–	–	1579	0.43	–	–	1580	1.28
C15H24O	Cariophyllene oxide	1582	1585	1.99	1585	2.11	–	–	1586	2.64
Total identified (%)	75.0		82.5		81.4		70.8
Not identified %	25.0		17.5		18.6		29.2

OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer; KI_L, Kovats index from literature; KI_C, Kovats index calculated; %, relative percentage.

Table 3. Antioxidant activity of the essential oils from the leaves of L. thymoides.

Sample	Scavenging of DPPH (EC50, µg/mL)	β-Carotene bleaching test (%AA)
OAA	> 243.0	10.8 ± 0.9
OJW	> 243.0	3.2 ± 0.2
OOS	> 243.0	3.2 ± 1.0
OJS	236.5 ± 3.5	0.3 ± 0.3
AAc	3.8 ± 1.8	15.1 ± 6.7
BHT	18.0 ± 1.0	NT
BHA	1.8 ± 0.2	NT
QUE	NT	45.4 ± 12.7
PYR	NT	2.5 ± 1.3

Data were expressed as mean ± SEM. EC₅₀, concentration that caused 50% of the DPPH scavenging; %AA, percentual of antioxidant activity in the β-carotene bleaching assay; OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer; AAc, ascorbic acid; BHT, butylated hydroxytoluene; BHA, butylated hydroxyanisole; QUE, quercetin; PYR, pyrogallol; NT, non tested.

Table 4. Antimicrobial activity of the essential oils from leaves of L. thymoides.

	Sample
	OAA		OJW		OOS		OJS		CHL	NYS
Microorganism	MIC	MMC	MIC	MMC	MIC	MMC	MIC	MMC	MIC	MIC
E. coli CCMB 261	R		R		R		R		0.675	NT
S. aureus CCMB 262	0.004	0.53	10.19	10.19	<0.004	0.26	<0.004	>8.43	0.003	NT
P. aeruginosa CCMB 268	R		NT		R		R		0.084	NT
S. cholerasuis CCMB 281	8.43	8.43	NT		R		R		0.005	NT
B. cereus CCMB 282	R		NT		R		R		0.005	NT
M. luteus CCMB 283	0.03	8.43	NT		0.07	>8.43	0.03	>8.43	0.003	NT
C. albicans CCMB 286	<0.004	4.21	R		<0.004	4.21	<0.004	8.43	NT	1.25

MIC, minimum inhibitory concentration in mg/mL; MMC, minimum microbicidal concentration in mg/mL; OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer; CHL, chloramphenicol; NYS, nystatin; R, resistant; NT, non-tested.

Figure 1. Vasorelaxant effect of essential oils from leaves of L. thymoides on tonic contractions induced by phenylephrine (Phe) 1.0 µM in isolated rat rings aorta. (A) Contractions induced in the presence of functional endothelium; (B) contractions induced in the absence of functional endothelium; (C) EC₅₀ values in the presence (E+) or absence (E−) of endothelium; (D) E_max values obtained in the presence (E+) or absence (E−) of endothelium. Data are presented as mean ± SEM (n = 5); *EC₅₀ or E_max values that were statistically different (t-test, p < 0.05) when compared with values of the same oil in the presence (E+) or absence (E−) of functional endothelium. OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer.

Figure 2. Spasmolytic effect of essential oils from leaves of L. thymoides on the tonic contractions induced with KCl 60 mM in isolated rat uterus. (A) Concentration–response curves of essential oils; (B) EC₅₀ values of essential oils. Data are presented as mean ± SEM (n = 5); *EC₅₀ values that were statistically different (ANOVA, p < 0.05). OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer.

Figure 3. Effect of essential oils from leaves of L. thymoides on the tonic contractions induced with carbachol (CCh) 1.0 µM in isolated guinea-pig rings trachea. Data are presented as mean ± SEM (n = 5). OAA, essential oil from L. thymoides collected in autumn; OJW, essential oil from L. thymoides collected in winter; OOS, essential oil from L. thymoides collected in spring; OJS, essential oil from L. thymoides collected in summer.