Antimalarial and Erythrocyte Membrane Stability Properties of Globimetula braunii (Engle Van Tiegh) Growing on Cocoa in Plasmodium berghei-Infected Mice

Figures & data

Table 1 Quantitative Phytochemical Determination of Extract and Fractions of G. braunii

Phytochemical Contents	Solvent	Concentration (mg/mL)
		50	100	200
	ME	4.4±0.1	5.4±0.1	5.6±0.0
	HF	3.5±0.2*	4.6±0.1	5.7±0.1
Alkaloid	DF	5.8±0.1*	5.7±0.0	5.7±0.0
	EF	5.1±0.0*	5.5±0.0	5.7±0.0
	MF	4.8±0.1	5.6±0.0	5.8±0.1
	ME	3.0±0.0*	5.5±0.2	8.9±1.3*
	HF	1.8±0.0*	2.1±0.1	2.2±0.5*
Phenolic content	DF	4.1±0.0*	3.3±0.7	4.1±0.2
	EF	3.9±0.2*	5.5±0.1	8.4±0.0*
	MF	3.2±0.0*	4.5±0.0	6.6±0.3*
	ME	342.5±17.7	380±35.4	430.25±26.5
	HF	555±0.0*	530±35.4*	570.5±17.7*
Tanin content	DF	492.5±53	467.5±17.7	515.5±17.7
	EF	505±0.0*	530±35.4*	580.25±17.7*
	MF	155±70.7*	217.5±53.0*	267.5±44.2*
	ME	5.1±1.4*	8.5±0.4*	6.6±1.2
	HF	1.4±0.2*	2.8±0.1*	2.1±0.2*
Total flavonoids	DF	2.5±0.6	4.1±0.7	8.2±2.3*
	EF	4.7±0.4	3.0±0.2	8.0±0.9*
	MF	5.8±0.3*	7.2±0.7*	7.7±0.3*
	ME	67.4±8.6*	57.6±1.5*	64.6±0.8*
	HF	61.6±3.2*	57.6±1.7*	65.4±0.5*
Total saponin	DF	53.5±1.1*	60.2±2.3*	56.9±7.0*
	EF	34.6±1.1	46.5±14.7	40.1±12.6
	MF	36.2±20.3	49.3±11.0	29.4±12.4
	ME	7±1.4**	8.5±2.1**	10±2.8**
	HF	9±1.4**	14.5±0.7**	15±1.4**
Total terpenes	DF	7±1.4**	8.5±2.1**	11±1.4**
	EF	6±1.4*	7±1.4*	9.5±2.1*
	MF	1.5±0.7	2.5±0.7	3±1.41

Note: *P<0.05, **P<0.01.

Figure 1 Percentage parasitemia (A–E) and percentage parasite clearance (F–J) of animals infected with a chloroquine-susceptible strain of P. berghei and treated with 100,200 and 400 mg/kg of extract and fractions of G. braunii.

Abbreviations: NC, Negative (Infected) Control; PC, positive control treated with P-alaxin; MC, Methanol crude; HF, Hexane; DF, Dichloromethane; EF, Ethylacetate; MF, Methanol fractions.

Table 2 Hematological Parameter of P. berghei-Infected Mice After Treatment with n-Hexane Fraction of G. braunii Leaf Extract

	Treatments	PCV (x 10)	HB	RBC	Platelet	WBC (10^3)	LYMP(*10)	NEUT(*10)	MN	EO
	NORMAL	3.9±0.1	13.3±0.4*	6.2±0.2*	4.3±0.4*	11.4±0.5*	8.9±3.0*	2.7±0.2	1±0	2.5±0.7
	P-ALAXIN	3.6±0	11.6±0.4	6.1±0.1	4.9±0.3	11.2±0.2	8.9±3.0	2.8±0.4	1±0	1±0
Full blood count (NK65-infected)	NEGATIVE	2.3±0.4	8.3±0.4	4.1±0.2*	4.4±0.5*	5.3±0.4*	5.4±0.6*1	3.7±0.3	1.5±0.7	1±0
	100mg/kg	2.4±0.1	7.1±0.1	3.7±0.1	6.8±0.4	7.1±0.1	6.3±1.0	3.9±0.4	1±0	1.5±0.7
	200mg/kg	4.1±0.1	13.5±0.1*	7.1±0.1*	7.2±0.3*	8.1±0.1*	7.6±0.6*	2.3±0.4	1±0	1.5±0.7
	400 mg/kg	3.7±0.2	11.15±0.2	5.4±0.2	5.8±0.4	9.5±0.2	7.8±2.2	3.4±0.3	1.5±0.7	0.5±0.7
	NORMAL	3.9±0.1	13.3±0.4*	6.3±0.2*	4.4±0.4	1.1±0.5*	3.9±2.7*	2.7±0.2*	1±0	2.5±0.7
	ARTEQUIN	2.9±0	9.3±0.4	4.4±0.1	5.7±0.3	0.8±0.2	3.8±0.6	2.8±0.4	1.5±0	1±0
Full blood count (ANKA-infected)	NEGATIVE	1.7±0.4	5.4±0.4	2.2±0.2	9±0.5	0.9±0.4	3.2±1.0	4.2±0.4	1±0.7	0.5±0
	100mg/kg	2.1±0.1	6.9±0.1*	3.1±0.1*	4.1±0.4	2.2±0.1*	4±0.6*	3.7±0.4*	1.5±0	1±0.7
	200mg/kg	2.25±0.1	5.6±0.1*	2.6±0.1*	6.3±0.3	1.7±0.1*	3.7±2.2*	4.0±0.4*	1.5±0	0.5±0.7
	400 mg/kg	1.9±0.2	6.5±0.2*	3.1±0.1*	4.8±0.4	1.8±0.2*	3.95±0.0*	3.5±0.3*	1±0.7	1±0.7

Note: *P<0.05.

Abbreviations: Neut, Neutrophils; Lym, Lymphocytes; Mono, Monocytes; Eosin, Eosinophils.

Table 3 Heme Contents of Mice Infected with Susceptible (NK 65) and Resistant (ANKA) Strains of P. berghei, Treated with n-Hexane Fraction of G. braunii

Treatment	HEME NK 65 (µmole/mL of Blood)	HEME ANKA (µmole/mL of Blood)
NORMAL	8.0±0.4	16.9±0.2
ARTEQUIN	10.3±0.6	16.8±0.1
NEGATIVE	8.7±0.3	14.8±0.6
100 mg/kg	9.8±1.0	16.6±0.3
200mg/kg	11.5±0.0	16.9±0.6
400mg/kg	6.9±0.5	17.6±0.4

Figure 2 Effects of n-hexane fraction of G. braunii on percentage parasitemia (a), parasite clearance (b) of mice infected with resistant P. berghei (ANKA), and hemozoin contents of mice infected with susceptible (NK 65) (c) and resistant (ANKA) (d) strains of P. berghei. *P<0.05, **P<0.01; ****P<0.0001.

Table 4 The Most Abundant Phytochemical Constituents of n-Hexane Fraction of G. braunii

S/N	Peak No	Compound Name	Abundance (Percentage)	Structure	R. Time	I.Time	F. Time
1	76	Friedelan-3-one Formula=C30H50O	27.68		32.267	32.175	32.425
2	68	Octamethyl, olean-12-en-3-one Formula=C30H48O	7.41		30.571	30.465	30.805
3	75	Epoxynaphthalene Formula= C23H36O2	5.32		32.137	32.075	32.175
4	65	Alpha-Amyrin Urs-12-en-3-ol Formula=C30H50O	3.59		30.043	29.905	30.175

Figure 3 The GC-MS analysis of n-hexane fraction showing the presence of friedelan-3-one (A) and the effects of n-hexane fraction of G. braunii on erythrocyte membrane stability using heat-induced (B) and hypotonic solution-induced (C) erythrocyte membrane distabilisation.