Isolation and identification of antioxidant and hyaluronidase inhibitory compounds from Ficus microcarpa L. fil. bark

Figures & data

Figure 1.  Separation scheme of bioactive compounds from F. microcarpa.

Figure 2.  HPLC chromatograms as monitored by UV absorption at 280 nm for fractions F1–F5 in 50% methanol, as obtained by Sephadex LH-20 column chromatography of the ethyl acetate extract of F. microcarpa bark. The peaks numbered with Arabic numerals in the order of increasing retention time correspond to the compounds labeled with the numerals in Figure 3.

Figure 3.  Chemical structures of isolated compounds from F. microcarpa.

Figure 4.  HPLC chromatograms as monitored by UV absorption at 280 nm (ODSpak column (150 mm × 4.60 mm; Shodex, Co.. Japan) for catechin, epicatechin, compounds 6, 7, and their hydrolyzates. (A) Standard of catechin and epicatechin (200 µg/mL); (B) compound 6; (C) hydrolyzate of 6; (D) compound 7; (E) hydrolyzate of 7. 1, catechin; 2, epicatechin.

Figure 5.  Hyaluronidase inhibitory activity by methanol extract and fractions of F. microcarpa. Each value is expressed as mean ± SD (n = 3), concentration of sample in assays is expressed as final concentration.

Figure 6.  Hyaluronidase inhibitory activity by purified compounds of F. microcarpa. Each value is expressed as mean ± SD (n = 3), concentration of sample in assays is expressed as final concentration.

Table 1.  Antioxidant activities of isolated compounds from F. microcarpa.

Compound	Antioxidant activity (EC50, µM)
	DPPH^a	PMS-NADH^b	ABTS^+c
Protocatechuic acid	40.3 ± 1.4	279.2 ± 2.8	19.6 ± 1.3
Chlorogenic acid	61.6 ± 0.9	65.8 ± 2.0	28.2 ± 1.0
Methyl chlorogenate	31.9 ± 1.1	54.3 ± 1.4	16.0 ± 0.4
Catechin	11.4 ± 0.5	67.4 ± 1.8	4.1 ± 0.5
Epicatechin	11.3 ± 1.0	71.0 ± 1.4	2.9 ± 0.4
Procyanidin B1	12.4 ± 0.7	79.4 ± 0.9	—
Procyanidin B3	14.0 ± 0.4	80.5 ± 1.7	—
Trolox	17.6 ± 0.7	722.0 ± 3.4	2.0 ± 0.3

Note. —, not determined; each value is expressed as mean ± SD (n = 3). Concentration of sample in assays is expressed as final concentration.

^aDPPH radical scavenging activity.

^bSuperoxide free radical scavenging activity.

^cABTS⁺ free radical scavenging activity.

Table 2.  Compound contents in F. microcarpa bark and ethyl acetate fraction.

Compound	Content (mg/g DW)
	Bark^a	Ethyl acetate fraction^b
Catechin	6.7 ± 0.6	220 ± 1.9
Epicatechin	5.0 ± 0.9	60 ± 1.5
Chlorogenic acid	6.7 ± 0.4	101 ± 1.2
Methyl chlorogenate	—	—
Protocatechuic acid	—	—
Procyanidin B1	—	—
Procyanidin B3	—	—

Note. —, not measured. Each value is expressed as mean ± SD (n = 3).

^aCompound content in dried powder of F. microcarpa bark.

^bCompound content in dried ethyl acetate fraction.