New compound from the aerial parts of Achillea millefolium

Figures & data

Figure 1. Structure new compound isolated from aerial parts of A.millefolium.

Table 1. New compounds and known from the aerial parts of A.millefolium.

	Treatment
Compound	Nano Zn oxide (2 mM)%	Nano Zn oxide (4 mM)%	RI	Identification methods
Known compounds
α-Pinene	6.45	7.45	932	MS, RI
Camphene	2.14	4.65	950	MS, RI
Sabinene	1.02	1.87	975	MS, RI
α-Phellandrene	0.07	0.41	1000	MS, RI
Limonene	2.36	4.53	1020	MS, RI
Trans-limonene oxide	0.25	1	1140	MS, RI
Borneol	20.14	36.35	1160	MS, RI
γ-Terpinene	1.14	1.45	1195	MS, RI
Carone	1.54	1.1	1200	MS, RI
Carvone	2.35	1.45	1240	MS, RI
Bornyl acetate	2	1.74	1281	MS, RI
Thymol	10.14	10.01	1300	MS, RI
Carvacrol	8.14	10.14	1308	MS, RI
β-caryophyllene	1.25	1.74	1418	MS, RI
γ-Elemene	1.08	1.01	1430	MS, RI
γ-Muurolene	0.74	0.1	1476	MS, RI
β-Selinene	0.87	0.41	1490	MS, RI
β-Guaiene	0.7	0.98	1496	MS, RI
α-Selinene	0.35	0.11	1490	MS, RI
Calarene	0.47	0.14	1500	MS, RI
Calacorene	0.74	0.35	1529	MS, RI
Ccaryophyllene oxide	1.05	0.21	1579	MS, RI
Torreyol	1.45	0.78	1640	MS, RI
Cadalene	2.14	0.61	1670	MS, RI
Yield	1.54	2.01
Total	63.67	83.15
New compound
Dihydro-a-cyclogeranyl hexanoate				UV, MS, MPLC,NMR

^aPercentage composition determined on column HP 5.

^bThe retention Kovats indices were determined on HP 5 capillary column in reference to n-alkanes.

MS: mass spectroscopy, RI: retention index.

Table 2. Effect of nano Zn oxide on extraction yields, total phenolic contents and total flavonoid contents of A.millefolium extracts.

	Extract	Extraction yield^a	Total phenolic^b	Total flavonoid^c
1	Control	66.14 ± 24	78.14 ± 32	80.24 ± 68
2	Nano Zn oxide (2mM)	68.47 ± 21	80.14 ± 78	84.24 ± 19
3	Nano Zn oxide (4mM)	100.14 ± 55	162.14 ± 57	121.47 ± 64

The data are expressed as mean ± SD.

^aExpressed as milligram of extract per gram dry material.

^bExpressed as milligram of gallic acid per gram dry extract.

^cExpressed as milligram of rutin per gram dry extract.

Table 3. Antimicrobial activity of the new compound and borneol (µg/mL) from A.millefolium using minimum inhibitory (MIC) and minimum bactericidal/fungicidal (MBC/MFC) test.

	Borneol	C1	Antibiotic (Streptomycin)
	MIC	MIC	MIC
Microorganisms	MBC	MBC	MBC
Bacillus cereus	7.0 ± 0.12	3.5±0.27	10±0.70
	7.0 ± 0.21	3.5 ± 0.41	15 ± 0.32
Enterococcus faecalis	7.5 ± 0.21	4.0 ± 0.28	10 ± 0.59
	7.0 ± 0.74	4.0 ± 0.17	10 ± 0.52
Staphylococcus aureus	20.0 ± 0.14	3.5 ± 0.01	250 ± 0.0
	20.0 ± 0.65	3.5 ± 0.05	250 ± 0.0
Escherichia coli	4.5 ± 0.31	2.5 ± 0.05	25 ± 0.5
	5.0 ± 0.52	2.5 ± 0.08	25 ± 0.5
Pseudomonas aeruginosa	25.0 ± 0.11	10 ± 0.74	100 ± 0.34
	20 ± 0.05	10 ± 0.24	150 ± 0.87
Proteus mirabilis	10.0 ± 0.54	4.5 ± 0.61	20 ± 0.54
	8.0 ± 0.12	4.0 ± 0.08	15 ± 0.5
Salmonella typhimurium	5.5 ± 0.24	4.5 ± 0.06	15 ± 0.05
	4.5 ± 0.14	4.5 ± 0.11	15 ± 0.02
Citrobacter freundii	6.5 ± 0.08	4.5 ± 0.17	15 ± 0.87
	6.0 ± 0.14	4.5 ± 0.20	20 ± 0.56
Microorganisms	Borneol	C1	Mycotic (fluconazole)
	MIC	MIC	MIC
	MFC	MFC	MFC
Candida albicans	4.5 ± 0.21	3.0 ± 0.35	4.5 ± 0.00
	4.5 ± 0.24	3.0 ± 0.21	5 ± 0.00
Aspergillus fumigatus	4.5 ± 0.65	3.5 ± 0.28	5 ± 0.9
	4.5 ± 0.84	3.0 ± 0.47	3.5 ± 0.5

All tests were performed in triplicate, repeated three times twice (means ± S.D). Statistically, the differences were significant at p ≤ 0.01 or at least at p ≤ 0.05 (in few cases), using Duncan test. C1: dihydro-a-cyclogeranyl hexanoate.

Table 4. Antioxidant activity new compound and borneol: scavenging activity (expressed as IC₅₀ values: µg/mL), and b-carotene bleaching test. Reducing power was expressed as EC₅₀ values (µg/mL). Butylhydroxyanisole (BHA) and ascorbic acid were used as positive controls.

Tested compounds	IC50 (μg/mL)
A.millefolium EO	12.97 ± 0.14 µg/mL
c1	11.79 ± 0.21 µg/mL
borneol	12.11 ± 0.60 µg/mL
A.millefolium EO (b–Carotenes IC50 µg/ml)	12.47 ± 0.28 µg/mL
c1	11.94 ± 0.74 µg/mL
A.millefolium EO (PR EC50 µg/ml)	12.97 ± 0.21 µg/mL
c1	11.65 ± 0.05 µg/mL
BHA	13.87 ± 0.22 µg/mL
AA	12.97 ± 0.11 µg/mL

Values are mean ± SD of three replications.

*IC₅₀ values have been presented with their respective 95% confidence limits.

Table 5. Components identified and their antioxidant activity relative percentages.

Compounds	%
c1	45.14
Borneol	21.11
Other components	36.68

Results are expressed as a percentage of antioxidant activity relative. Experiments were carried out in triplicate.

Table 6. Lipid peroxidation inhibition of new component and borneol (expressed as IC₅₀ values: µg/ml). Trolox and ascorbic acid (AA) were used as positive controls.

Tested compounds	IC50 (μg/mL)
A.millefolium EO	78.54 ± 0.21 µg/mL
C1	80.41 ± 0.24 µg/mL
Borneol	75.60 ± 0.34 µg/mL
Trolox	72.18 ± 0.35 µg/mL
AA	75.61 ± 0.21 µg/mL

Experiments were carried out in triplicate and the results are expressed as mean ± SD.

Table 7. Effects of A.millefolium EO (45.0 μg/mL), new component and borneol (45.0 μM) on NO production in LPS-stimulated RAW-264.7 macrophages.

Tested compounds	NO inhibition (%)
A.millefolium EO	78 ± 0.21
C1	88 ± 0.05
Borneol	75 ± 0.04
L-NAME	73 ± 0.05

Results are expressed as a percentage of nitrite production by control cells maintained in culture medium. Experiments were carried out in triplicate and the results are expressed as mean ± SD.