Acetylcholinesterase and butyrylcholinesterase inhibitory activity of Pinus species essential oils and their constituents

Figures & data

Table 1.  Chemical composition of the essential oils from P. nigra var. calabrica (P1), P. heldreichii subsp. leucodermis (P2), and P. nigra subsp. nigra (P3).

I^a/I^b	Compound	ID method^d	%^c
			P1	P2	P3
936/1032	α-Pinene	I, MS, Co-GC	24.6	24.2	25.3
953/1076	Camphene	I, MS	1.1	0.4	2.2
973/1138	Sabinene	I, MS, Co-GC	0.2	0.1	12.8
978/1118	β-Pinene	I, MS, Co-GC	10.9	8.4	4.8
1005/1186	α-Phellandrene	I, MS	0.4	—	0.3
1012/1159	δ-3-Carene	I, MS	1.7	0.2	—
1016/1188	α-Terpinene	I, MS, Co-GC	0.2	1.5	0.4
1025/1280	p-Cymene	I, MS	—	0.2	—
1030/1218	β-Phellandrene	I, MS, Co-GC	6.3	tr	tr
1032/1203	Limonene	I, MS, Co-GC	tr	7.8	22.6
1040/1246	(Z)-β-Ocimene	I, MS	4.0	—	—
1047/1266	(E)-β-Ocimene	I, MS	—	3.7	1.7
1059/1255	γ-Terpinene	I, MS, Co-GC	0.3	1.0	0.8
1089/1290	Terpinolene	I, MS, Co-GC	1.1	5.9	4.5
1098/1553	Linalool	I, MS, Co-GC	0.1	0.5	—
1100/1400	n-Nonanal	I, MS	0.1	0.1	tr
1113/	endo-Fenchol	I, MS	tr	0.1	—
1128/1499	α-Campholene aldehyde	I, MS	0.1	0.3	0.1
1138/1665	trans-Pinocarveol	I, MS	tr	0.2	—
1178/1611	Terpinen-4-ol	I, MS, Co-GC	0.1	0.8	0.4
1189/1683	α-Terpineol	I, MS	0.3	1.7	8.3
1228/	Myrtenyl acetate	I, MS	0.2	0.6	0.1
1240/	Linalyl acetate	I, MS, Co-GC	tr	3.6	tr
1265/1604	Thymol methyl ether	I, MS	0.4	0.2	—
1289/1597	Bornyl acetate	I, MS	0.7	2.7	0.2
1350/1466	α-Cubebene	I, MS	1.0	7.6	—
1370/1733	Neryl acetate	I, MS	0.3	—	—
1388/1765	Geranyl acetate	I, MS	0.4	0.4	—
1373/1192	α-Ylangene	I, MS	—	—	0.6
1377/1497	α-Copaene	I, MS	0.3	0.4	tr
1385/1535	β-Bourbonene	I, MS	—	0.5	—
1403/	α-Bergamotene	I, MS	1.0	—	—
1408/2030	Methyl eugenol	I, MS	0.5	0.3	—
1418/1612	trans-Caryophyllene	I, MS, Co-GC	4.2	4.5	3.0
1439/1628	Aromadendrene	I, MS	0.4	—	—
1441/1662	trans-β-Farnesene	I, MS	—	0.9	—
1454/1690	α-Humulene	I, MS, Co-GC	1.2	1.0	0.5
1478/1725	Germacrene D	I, MS	0.8	0.7	0.5
1483/	γ-Muurolene	I, MS	0.9	0.2	0.1
1495/1731	Epi-bicyclosesquiphellandrene	I, MS	—	—	0.7
1499/1742	α-Muurolene	I, MS	—	—	0.8
1512/	Bicyclo[4.4.0]dec-1-en, 2-isopropyl-5-methyl-9-methylene	I, MS	0.7	0.3	0.2
1515/1765	γ-Cadinene	I, MS	9.9	1.0	0.7
1524/1772	δ-Cadinene	I, MS, Co-GC	1.9	1.8	0.5
1534/2496	Ethyl laurate	I, MS	0.7	0.3	0.4
1567/2502	Lauric acid	I, MS	1.7	0.8	0.5
1612/	Tetradecanal	I, MS	0.3	0.5	0.2
1624/	Junipene	I, MS	—	—	0.1
1722/	Farnesol	I, MS	0.4	—	tr
1810/	Hexadecanal	I, MS	1.5	0.3	0.1
1881/	1-Octadecene	I, MS	0.3	0.2	0.3
1893/	1-Nonadecene	I, MS	0.9	—	—
1934/	Methyl palmitate	I, MS, Co-GC	—	0.8	—
1942/	(E)-3-Cembrene A	I, MS	0.9	—	0.3
1960/	Sandaracopimaradiene	I, MS	—	0.8	—
1969/2931	Palmitic acid	I, MS, Co-GC	—	0.5	—
1987/2713	Myristic acid	I, MS, Co-GC	0.8	0.2	0.4
1996/	Methyl linoleate	I, MS	1.2	—	—
1989/2375	Manoyl oxide	I, MS	6.2	0.6	—
2000/2000	Eicosane	I, MS	0.7	—	—
2022/	Octadecanal	I, MS	0.2	—	—
2100/2100	Heneicosane	I, MS, Co-GC	—	0.5	0.3
2300/2300	Tricosane	I, MS, Co-GC	0.5	0.2	0.6
2365/	Methyl neoabietate	I, MS	tr	0.6	—
2500/2500	Pentacosane	I, MS, Co-GC	0.8	0.4	—
2900/2900	Nonacosane	I, MS, Co-GC	—	0.8	0.4
	Identified compounds		93.4	91.3	95.7

Note. Hydrodistillation (HD), cold-pressing (CP), supercritical fluid extraction (SFE).

^aRetention index on SE30 MS column.

^bRetention index on HP-Innowax column.

^cMean value ± standard error, n = three independent determinations. Compositional values less than 0.1% are denoted as traces (tr).

^dI, Retention index; MS, mass spectrum; Co-GC: co-injection with authentic compound.

Figure 1.  Dose-dependent inhibitory activity of Pinus essential oils against AChE (a) and BChE (b). Data are given as mean ± SD (n = 3).

Figure 2.  Dose-dependent inhibitory activity of terpinolene and β-phellandrene against AChE. Data are given as mean ± SD (n = 3).

Figure 3.  Dose-dependent inhibitory activity of terpinolene, β-phellandrene, terpinen-4-ol, linalyl acetate, and trans-caryophyllene against BChE. Data are given as mean ± SD (n = 3).

Table 2.  Cholinesterase inhibitory activity of essential oils and identified constituents from Pinus species (IC₅₀, μg/mL).

	AChE	BChE	SI (BChE/AChE)
Essential oils
P. heldreichii subsp. leucodermis	51.1 ± 1.8*	80.6 ± 2.3*	1.6
P. nigra var. calabrica	101.5 ± 2.4*	128.0 ± 2.4*	1.3
P. nigra subsp. nigra	94.4 ± 1.8*	162.5 ± 1.4*	1.7
Terpenes
β-Phellandrene	120.2 ± 2.0*	> 50
Terpinolene	156.4 ± 2.8*	147.1 ± 2.5*	0.9
Terpinen-4-ol	21.4% (1.2 mM)^Citation21	107.6 ± 1.2*
Linalyl acetate	38% (82 μg/mL)^Citation35	168.7 ± 2.1*
trans-Caryophyllene	32% (0.06 mM)^Citation22	78.6 ± 1.3*
Positive controls
Physostigmine	0.1 ± 0.01	0.2 ± 0.01	2
Galanthamine hydrobromide	0.3 ± 0.04	3.0 ± 0.04	10

Note. IC₅₀ values are mean ± SD (n = 3). One-way ANOVA analysis: ***p < 0.0001; Dunnett’s test: *p < 0.01.

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