Bioassay-guided isolation of anti-inflammatory and antinociceptive compound from Plumbago zeylanica leaf

Figures & data

Table 1.  Preliminary phytochemical studies and percentage yields of various extracts and fractions of P. zeylanica.

Plant extract	Yield (%, w/w)	Constituents
Pz.L-P	2.83	Fats, steroids, and naphthaquinone
Pz.L-C	2.73	Steroids and naphthaquinone
Pz.L-A	1.2	Tannins, flavonoids, triterpenoids, and naphthaquinone
Pz.L-E	14.26	Carbohydrates, tannins, flavonoids, and saponins
Pz.L-W	22.2	Carbohydrates, tannins, alkaloids, flavonoids, and saponins
F1	26.69	Tannins
F2	45.81	Tannins, flavonoids, and triterpenoids
F3	2.18	Plumbagin
F4	21.43	Triterpenoids

Pz.L, P. zeylanica leaves; P, petroleum ether (60–80°); C, chloroform; A, acetone; E, ethanol; W, aqueous; F₁, fraction 1; F₂, fraction 2; F₃, fraction 3; F₄, fraction 4.

Table 2.  Anti-inflammatory activity of various extracts of P. zeylanica.

Treatment	Dose (mg/kg)	Average volume of mercury displaced (mL)
		0 h	1 h	2 h	3 h	4 h	5 h
Control	–	0.64 ± 0.02	0.93 ± 0.02	0.90 ± 0.00	1.13 ± 0.03	1.08 ± 0.04	0.93 ± 0.04
Diclofenac	100	0.56 ± 0.03	0.82 ± 0.04 (11.8)	0.68 ± 0.00* (24.4)	0.79 ± 0.00** (30.3)	0.67 ± 0.02** (37.9)	0.56 ± 0.06** (39.78)
Pz.L-P	200	0.56 ± 0.03	0.84 ± 0.03 (9.6)	0.82 ± 0.03 (8.8)	0.90 ± 0.02 (20)	0.92 ± 0.03 (14.8)	0.86 ± 0.03 (7.5)
	400	0.56 ± 0.02	0.82 ± 0.07 (11.8)	0.83 ± 0.03 (7.7)	0.78 ± 0.05** (30.9)	0.91 ± 0.05 (15.7)	0.71 ± 0.02* (23.6)
Pz.L-C	200	0.54 ± 0.02	0.83 ± 0.01 (10.7)	0.84 ± 0.05 (6.6)	0.97 ± 0.01 (14.1)	0.93 ± 0.02 (13.8)	0.80 ± 0.02 (13.9)
	400	0.53 ± 0.01	0.84 ± 0.05 (9.6)	0.80 ± 0.04 (11.1)	0.96 ± 0.04 (15)	0.92 ± 0.02 (14.8)	0.74 ± 0.07 (20.4)
Pz.L-A	200	0.58 ± 0.04	0.78 ± 0.02 (16.1)	0.79 ± 0.04 (12.2)	1.00 ± 0.01* (11.5)	0.90 ± 0.02*(16.6)	0.79 ± 0.01 (23.6)
	400	0.60 ± 0.03	0.76 ± 0.01 (18.2)	0.74 ± 0.02* (17.2)	0.74 ± 0.05** (34.51)	0.70 ± 0.04** (35.18)	0.60 ± 0.01** (35.48)
Pz.L-E	200	0.54 ± 0.03	0.84 ± 0.03 (9.6)	0.81 ± 0.03 (10)	0.96 ± 0.03 (15)	0.94 ± 0.02 (12.9)	0.93 ± 0.04 (0)
	400	0.58 ± 0.00	0.88 ± 0.03 (5.3)	0.82 ± 0.03 (8.8)	0.98 ± 0.06 (13.2)	0.90 ± 0.06 (16.6)	0.78 ± 0.04 (16.1)
Pz.L-W	200	0.54 ± 0.02	0.84 ± 0.03 (9.6)	0.81 ± 0.05 (10)	0.98 ± 0.04 (13.2)	0.99 ± 0.00 (8.3)	0.86 ± 0.03 (7.5)
	400	0.56 ± 0.01	0.80 ± 0.01 (13.9)	0.80 ± 0.03 (11.1)	0.90 ± 0.04 (20)	0.93 ± 0.02 (13.8)	0.77 ± 0.05 (17.2)

Each value represents the mean ± SEM (n = 6); *p < 0.05, **p < 0.01 vs. control (Dunnett’s test). Values in parenthesis represents percentage inhibition.

Table 3.  Anti-inflammatory activity of various fractions of acetone extract of P. zeylanica leaf.

Treatment	Dose (mg/kg)	Average volume of mercury displaced (mL)
		0 h	1 h	2 h	3 h	4 h	5 h
Control		0.60 ± 0.02	0.74 ± 0.04	0.92 ± 0.02	1.04 ± 0.03	1.12 ± 0.03	1.05 ± 0.03
Diclofenac	100	0.57 ± 0.02	0.66 ± 0.03 (10.81)	0.69 ± 0.01** (25)	0.73 ± 0.03** (28.80)	0.77 ± 0.01** (31.25)	0.64 ± 0.02** (39.04)
F1	200	0.56 ± 0.01	0.63 ± 0.01 (14.86)	0.98 ± 0.02 (-6.52)	0.91 ± 0.05 (12.5)	1.02 ± 0.01 (8.92)	0.99 ± 0.06 (5.71)
F2	200	0.56 ± 0.01	0.65 ± 0.02 (12.16)	0.80 ± 0.04* (13.04)	0.88 ± 0.06* (15.38)	0.96 ± 0.06* (14.28)	0.87 ± 0.07* (17.14)
F3	2	0.55 ± 0.02	0.63 ± 0.01(14.86)	0.73 0.03** (20.65)	0.70 ± 0.04** (32.69)	0.79 ± 0.02** (29.46)	0.67 ± 0.01** (36.19)
F4	200	0.62 ± 0.01	0.68 ± 0.01 (8.10)	0.90 ± 0.06 (2.17)	0.89 ± 0.03 (14.42)	0.99 ± 0.06 (11.60)	0.96 ± 0.06 (8.57)

Each value represents the mean ± SEM (n = 6); *p < 0.05, **p < 0.01 vs. control (Dunnett’s test). Values in parenthesis represents percentage inhibition.

Table 4.  Effect of various extracts of P. zeylanica in mice on hot plate reaction time.

Treatment	Dose (mg/kg)	15 min	30 min	60 min	120 min
Control	2 ml/kg	4.83 ± 0.47	5.33 ± 0.56	5.50 ± 0.56	5.50 ± 0.62
Diclofenac	100	5.42 ± 0.47 (12.21)	8.58 ± 0.42** (60.97)	14.56 ± 1.20** (164.7)	20.99 ± 0.97** (281.63)
Pz.L-P	200	4.33 ± 0.88 (−10.35)	5.83 ± 0.47 (9.38)	7.33 ± 0.66 (33.27)	6.00 ± 0.36 (24.18)
	400	4.16 ± 0.83 (−13.87)	5.33 ± 0.33 (0)	6.83 ± 0.74 (24.18)	5.50 ± 0.22 (0)
Pz.L-C	200	4.33 ± 0.80 (−10.35)	5.16 ± 0.47 (3.01)	6.67 ± 0.84 (21.27)	6.67 ± 0.21 (21.27)
	400	4.00 ± 0.44 (−17.18)	5.00 ± 0.73 (6.19)	6.16 ± 1.07 (12.00)	6.50 ± 0.67 (18.18)
Pz.L-A	200	4.50 ± 0.67 (6.83)	4.50 ± 0.42 (−15.57)	5.22 ± 0.18 (−5.09)	5.83 ± 0.16 (5.66)
	400	5.00 ± 0.36 (3.51)	4.83 ± 0.37 (9.38)	6.67 ± 0.84 (21.27)	6.16 ± 0.83 (10.71)
Pz.L-E	200	4.16 ± 0.47 (−13.87)	4.80 ± 0.42 (9.94)	5.05 ± 0.45 (−8.08)	5.83 ± 0.79 (5.66)
	400	5.66 ± 0.21 (−17.18)	5.00 ± 0.36 (6.19)	7.00 ± 0.68 (27.27)	5.66 ± 0.49 (2.90)
Pz.L-W	200	4.16 ± 0.70 (−13.87)	5.83 ± 0.30 (9.38)	5.66 ± 0.75 (2.90)	6.83 ± 0.47 (19.47)
	400	5.00 ± 0.73 (3.51)	4.66 ± 0.33 (−12.57)	6.83 ± 0.91 (24.18)	5.66 ± 0.49 (2.90)

Each value represents the mean ± SEM (n = 6); **p < 0.01 vs. control (Dunnett’s test).

Table 5.  Effect of various fractions of acetone extract of P. zeylanica leaf on hot plate reaction time in mice.

Treatment	Dose (mg/kg)	15 min	30 min	60 min	120 min
Control	–	6.66 ± 0.95	6.50 ± 1.25	6.50 ± 1.08	6.16 ± 1.13
Diclofenac	100	8.16 ± 1.04 (22.52)	8.16 ± 1.27 (25.53)	16.16 ± 1.90^** (148.6)	23.16 ± 1.16** (275.97)
F1	200	4.98 ± 0.83 (−25.22)	6.00 ± 1.54 (−7.69)	7.66 ± 1.30 (17.84)	4.66 ± 1.83 (−24.35)
F2	200	7.33 ± 1.05 (10.06)	6.50 ± 1.36 (0)	4.33 ± 0.84 (−33.38)	7.00 ± 1.12 (−13.63)
F3	2	9.00 ± 0.82* (35.13)	17.00 ± 2.30** (161.5)	23.33 ± 1.76** (258.92)	17.33 ± 1.42** (181.33)
F4	200	5.48 ± 0.82 (−17.77)	5.33 ± 1.04 (−18)	5.00 ± 0.68 (−23.07)	7.50 ± 1.66 (−8.08)

Each value represents the mean ± SEM (n = 6); *p < 0.05, **p < 0.01 vs. control (Dunnett’s test).

Table 6.  Effect of P. zeylanica extracts on formalin induced nociception in mice.

Treatment	Dose (mg/kg)	Duration of pain response (s)		Inhibition (%)
		First phase (0–5 min)	Second phase (15–30 min)	First phase	Second phase
Control	–	62.66 ± 2.98	177.83 ± 4.35	–	–
Diclofenac	100	61.66 ± 3.19	100.67 ± 6.76**	1.59	43.38
Morphine	10	30.89 ± 2.56**	40.86 ± 5.72**	50.7	77.02
Pz.L-P	200	94.83 ± 3.06	152.68 ± 4.87*	–51.34	14.14
	400	87.50 ± 8.74	141.00 ± 5.77**	–39.64	20.71
Pz.L-C	200	91.00 ± 7.16	171.67 ± 7.49	–45.22	3.46
	400	71.16 ± 6.6	149.33 ± 8.19	–13.56	16.02
Pz.L-A	200	58. 50 ± 4.78	30.16 ± 3.70**	6.63	83.03
	400	60.83 ± 2.41	30.66 ± 5.31**	2.92	82.75
Pz.L-E	200	70.66 ± 8.72	177.33 ± 4.33	–11.71	0.28
	400	75.66 ± 5.19	163.67 ± 6.46	–20.74	7.96
Pz.L-W	200	73.16 ± 6.93	180.80 ± 5.95	–16.75	–1.67
	400	68.00 ± 3.17	170.17 ± 2.16	–8.52	4.3

Each value represents the mean ± SEM (n = 6); *p < 0.05, **p < 0.01 vs. control (Dunnett’s test).

Table 7.  Effect of P. zeylanica fractions on formalin induced nociception in mice.

Treatment	Dose (mg/kg)	Duration of pain response (s)		Inhibition (%)
		First phase (0–5 min)	Second phase (15–30 min)	First phase	Second phase
Control	–	54.00 ± 4.78	177.00 ± 6.70	–	–
Diclofenac	100	47.16 ± 5.61	39.33 ± 4.63	12.66	77.77
Morphine	10	30.00 ± 3.40**	41.66 ± 2.70**	44.44	76.46
F1	200	54.33 ± 4.71	169.17 ± 6.96	0.60	4.44
F2	200	57.66 ± 6.83	61.16 ± 3.46**	6.77	65.44
F3	2	39.44 ± 4.63**	47.16 ± 5.54**	26.96	73.44
F4	200	53.50 ± 2.96	170.83 ± 3.50	0.74	3.48

Each value represents the mean ± SEM (n = 6); **p < 0.01 vs. control (Dunnett’s test).