The Research of Lygodium

Lygodium is the dry root and rhizome of Lygodium japonicum (Thunb.)Sw. which belongs to the family Lygodiaceae. Lygodium is the only genus of Lygodiaceae comprises 45 species throughout the world. In China, there are 10 species of Lygodium distributed in the southwest and south China, and five species of them had been used for Chinese herbs medicine to treat hepatitis and dysentery[1]. They are named Lygodium, Lygodium of hainan, Crankshaft Lygodium, Angustifolia Lygodium, Pinnately lobed Lygodium, Willowlike leaves Lygodium, Reticulata Lygodium, yunnan Lygodium, Lobular Lygodium, Palm leaf Lygodium.


Introduction
Lygodium is the dry root and rhizome of Lygodium japonicum (Thunb.)Sw.which belongs to the family Lygodiaceae.Lygodium is the only genus of Lygodiaceae comprises 45 species throughout the world.In China, there are 10 species of Lygodium distributed in the southwest and south China, and five species of them had been used for Chinese herbs medicine to treat hepatitis and dysentery [1] .They are named Lygodium, Lygodium of hainan, Crankshaft Lygodium, Angustifolia Lygodium, Pinnately lobed Lygodium, Willowlike leaves Lygodium, Reticulata Lygodium, yunnan Lygodium, Lobular Lygodium, Palm leaf Lygodium.

Chemical composition
Currently, the research on the active components in the Lygodium were done on the underground parts.We summarize the structure and classification of these compounds from lygodium.

Apparatus
Melting points were determined on an X4-A micro-melting point apparatus and were uncorrected.ESI-MS spectra were measured on an Agilent 1100 LC-MSD-Trap-SL, and HR-ESI-MS spectra were measured on an Bruker Dal-tonics MicroTOFQ.NMR spectra were measured on a Bruker ARX-600 and 300 NMR spectrometer with tetra-methylsilane (TMS) as the internal reference and chemical shifts are expressed with δ (ppm).UV spectra were recorded on a Shimadzu UV-2201 spectrometer.IR spectra were recorded on a Bruker IFS-55 spectrophotometer.TLC was performed on silica gel GF254 (10-40 lm; Qingdao,China).Separations were performed by Semiprep-HPLC named Shimadzu SPD-10A apparatus equipped with UV detector under ODS column (i.d. 10 mm 9 200 mm).

Physical data of the new compound 1
The new compound,yellow powder, melting point:193-194℃.The molecular formula was determined as C 14 H 14 O 3 by HR-ESI-TOF-MS (m/z 231.1004[M + H] + , calcd.231.1016), along with 1 H-NMR and 13 C-NMR data.The UV spectrum displayed absorption bands at 207, 267 and 347 nm, closely resembling that of 1,4-naphthoquinones.The 13 C-NMR spectrum revealed 14 carbon resonances; in the low field area of it, two were assigned as carbonyl carbons, eight were assigned as aromatic carbons.However, in the high field area of 13 C-NMR spectrum, there were four carbon resonances all that assigned as sp3 carbons.By observing these data of assigned by analysis of the HSQC spectrum (Table 1).

Extraction and isolation
The air-dried roots of L. japonicum (Thunb.)Sw were crushed and extracted twice using reflux with 70% ethanol; the solution was concentrated under reduced pressure to obtain the residue, and then the residue was extracted with MeOH.The MeOH-soluble fraction (100 g) was isolated by column chromatography on silica gel and gradient elution with CHCl 3 :MeOH (50 : 1 to 1 : 1) gave 14 fractions.Fraction 8 was isolated by semipreparative ODS column using MeOH:H 2 O (65 : 35) as eluent to afford the new compound 2 (13mg).

Extraction and isolation
The air-dried roots of L. japonicum (Thunb.)Sw. (4 kg) were crushed and extracted twice under reflux with 70% EtOH.The solution was concentrated under reduced pressure to obtain the residue, and then the residue was extracted with MeOH.The MeOH-soluble fraction (100 g) was isolated by column chromatography on silica gel using gradient elution with CHCl 3 -MeOH (50:1 to 1:1), which gave 14 fractions.Fraction 9 (10 g) was subjected to silica gel column chromatography using CHCL 3 -MeOH(40:1 to1:1) in gradient to give fractions 1-4.Fraction 4 (3.7 g) was chromatographed on an ODS column eluting with MeOH-H 2 O system, giving two fractions.Fraction 2 (1.3 g) was isolated by a semipreparative ODS column using MeOH-H 2 O (65:35) as the eluent to afford lygodium A (9 mg) and a kown compound ponastteroside A(30 mg), respectively.

Apparatus
Melting points were determined on an X4-A micro-melting point apparatus and were uncorrected.ESI-MS spectra were measured on an Agilent1100 LC-MSD-Trap-SL, and HR-ESI-MS spectra were measured on a Bruker Daltonics MicroTOFQ.NMR spectra were measured on a Bruker ARX-600 NMR spectrometer with tetramethylsilane (TMS) as the internal reference and chemical shifts are expressed with δ (ppm).UV spectra were recorded on a Shimadzu UV-2201 spectrometer.IR spectra were recorded on a Bruker IFS-55 spectrophotometer.TLC was performed on silica gel GF254 (10-40 l; Qingdao, China).Separation was performed by semiprep HPLC using Shimadzu SPD-10A apparatus equipped with a UV detector under an ODS column (i.d. 10 mm * 200 mm).

Analysis and conclusions of the new compound 3:
The new compound 3,white powder, mp.245-246℃, gave positive response to Liebermann-Burchard reaction and Molish reaction, suggesting a steroid glycoside structure.The sugar was identified as glucose by co-TLC with authentic sample after acid hydrolysis.

Fig. 16 .
Fig. 16.The key correlations of the new compound 3.

New compounds from Lygodium japonicum 3.1 New naphthalene ketone from the root of Lygodium japonicum [5] 3.1.1 Extraction and isolation
Air-dried roots of L. japonicum (Thunb.)Sw.(4 kg) were crushed and extracted