Anticholinesterase, antioxidant, analgesic and anti-inflammatory activity assessment of Xeranthemum annuum L. and isolation of two cyanogenic compounds

Figures & data

Figure 1. Isolation scheme of compounds XA-1 and XA-2.

Figure 2. Structures of zierin (XA-1) and zierin xyloside (XA-2).

Table 1. ¹H and ¹³C NMR data for zierin (XA-1) and zierin xyloside (XA-2).

		Zierin (XA-1)		Zierin xyloside (XA-2)
		δC (ppm)	δH (ppm), J (Hz)	δC (ppm)	δH (ppm), J (Hz)
C/H atom
Aglycon
1	C	118.6		118.7
2	CH	69.9	5.97 s	68.3	5.89 s
3	C	133.6		132.9
4	CH	117.1	6.63 φ	116.7	6.65 φ
5	C	160.3		160.4
6	CH	120.4	6.56 φ	121.4	6.59 φ
7	CH	127.2	7.38 φ	126.2	7.43 φ
8	CH	128.8	7.43 φ	128.9	7.47 φ
Glucose
1′	CH	104.3	4.66 d (7.9)	104.1	4.51 d (7.9)
2′	CH	75.7	3.00–3.43 φ	76.3	3.00–3.45 φ
3′	CH	76.8	3.41 φ	77.1	3.45 φ
4′	CH	70.7	3.00–3.30 φ	70.4	3.00–3.30 φ
5′	CH	76.2	3.17 φ	75.9	3.25 φ
6′	CH2	62.5	3.55 φ	69.3	3.56 φ
			3.95 φ		3.84 φ
Xylose
1″	CH			106.5	4.39 d (7.6)
2″	CH			75.7	3.01 dd (7.6/9.0)
3″	CH			78.0	3.25 φ
4″	CH			70.9	3.36 m
5″	CH2			67.0	3.74 φ
					3.09 φ

φUnclear splitting for this peak.

Figure 3. Inhibition (%±SEM) of the extracts of Xeranthemum annuum (XA) against BChE at 100 μg mL⁻¹ (F, flower, R-S, root and stem; Gal, galanthamine – reference).

Figure 4. Metal-chelating capacity (%±SEM) of the extracts of Xeranthemum annuum (XA) at 100 μg mL⁻¹ (F, flower, R-S, root and stem; EDTA, ethylenediaminetetraacetic acid – reference).

Picture 1. Xeranthemum annuum L. (photo taken by Ilkay Erdogan Orhan).

Table 2. Total phenol and flavonoid amounts, DPPH radical scavenging and FRAP activities of the extracts of XA at 1000 μg mL−1.

Extracts	Total phenolcontenta±SEMb	Total flavonoidcontentc±SEM	Radical scavengingeffect (%±SEM) against DPPH	FRAP (absorbance at700 nmd±SEM)
XA-F-hexane	NTe	NT	NT	NT
XA-F-CHCl3	–^f	9.16 ± 2.98	1.33 ± 0.21****	0.339 ± 0.006****
XA-F-EtOAc	14.36 ± 1.17	6.14 ± 0.52	3.84 ± 1.05****	0.312 ± 0.077****
XA-F-EtOH	71.27 ± 0.01	14.55 ± 1.03	13.37 ± 0.31****	0.565 ± 0.022****
XA-F-H2O	68.38 ± 0.58	12.36 ± 0.26	19.58 ± 3.66****	0.276 ± 0.005****
XA-R-S-hexane	5.28 ± 0.02	25.08 ± 4.12	2.74±±0.93****	0.402 ± 0.029****
XA-R-S-CHCl3	76.63 ± 1.08	62.23 ± 1.68	2.66 ± 0.42****	0.325 ± 0.026****
XA-R-S-EtOAc	46.52 ± 3.00	40.36 ± 6.99	17.06 ± 0.52****	0.585 ± 0.033****
XA-R-S-EtOH	145.51 ± 3.67	30.84 ± 1.56	77.33 ± 1.99**	1.126 ± 0.021***
XA-R-S-H2O	173.56 ± 2.16	1.37 ± 0.02	23.93 ± 0.42****	0.745 ± 0.031***
Gallic acid (reference for DPPH scavenging activity)			92.88 ± 0.19
Chlorogenic acid (reference for FRAP)				3.618 ± 0.01

** p < 0.01;

*** p < 0.001,

**** p < 0.0001.

^a Data expressed in mg equivalent of gallic acid to 1 g of extract.

^b Standard error mean (n = 3).

^c Data expressed in mg equivalent of quercetin to 1 g of extract.

^d Higher absorbance indicated the greater antioxidant activity.

^e Not tested.

^f Not able to calculate to very low absorbance.

Table 3. Analgesic effect of the R-S extracts of XA in PBQ-induced writhing model in mice.

Extracts	Dose (mg kg^−1)	Writhing number ± SEM^a	Inhibition rate (%)	Ulceration rate
Control		41.7 ± 4.2
XA-R-S-hexane	100	36.9 ± 4.5	11.5	0/6
	200	32.8 ± 3.1	21.3	0/6
XA-R-S-CHCl3	100	33.4 ± 3.6	19.9	0/6
	200	31.1 ± 2.9	25.4*^b	1/6
XA-R-S-EtOAc	100	42.6 ± 6.0	–^c	2/6
	200	39.5 ± 4.5	5.3	3/6
XA-R-S-EtOH	100	43.2 ± 5.1	–	0/6
	200	35.8 ± 2.7	14.1	0/6
XA-R-S-H2O	100	38.5 ± 3.2	7.7	0/6
	200	45.2 ± 4.6	–	0/6
ASA^d	100	21.5 ± 2.1	48.4***	4/6
	200	19.6 ± 1.7	52.9***	6/6

^a Standard error mean.

^b *p< 0.05,

*** p< 0.001 significant from the control.

^c No activity.

^d Acetyl salicylic acid (reference).

Table 4. Anti-inflammatory effect of the R-S extracts of XA in carrageenan-induced paw oedema model in mice.

		Swelling thickness (×10^−2 mm)±SEM^a (inhibition %)
Extracts	Dose (mg kg^−1)	90 min	180 min	270 min	360 min
Control		53.1 ± 5.3	60.4 ± 6.1	57.6 ± 5.6	66.3 ± 4.7
XA-R-S-hexane	200	49.9 ± 4.5 (6.0)	54.8 ± 5.7 (9.3)	59.6 ± 6.1	67.3 ± 6.0
XA-R-S-CHCl3	200	45.1 ± 3.7 (15.1)	49.6 ± 4.2 (17.9)	51.8 ± 4.6 (10.1)	57.1 ± 4.0 (13.9)
XA-R-S-EtOAc	200	57.5 ± 3.6	62.1 ± 3.9	65.2 ± 4.8	69.6 ± 5.3
XA-R-S-EtOH	200	54.4 ± 3.9	59.5 ± 3.6 (1.5)	55.8 ± 4.0 (3.1)	62.5 ± 5.2 (5.7)
XA-R-S-H2O	200	56.8 ± 6.4	65.1 ± 5.3	68.7 ± 5.9	72.5 ± 5.8
Indomethacin	10	40.2 ± 3.3 (24.3)*	44.1 ± 3.1 (26.9)*	39.7±±3.4 (31.1)**	40.8 ± 3.0 (38.5)**

^a Standard error mean.

^b *p < 0.05,

** p < 0.01, significant from the control.