Isolation of Prenylated Flavonoid from Ethyl Acetate Fraction of ...

Asian Transactions on Basic and Applied Sciences (ATBAS ISSN: 2221-4291) Volume 02 Issue 06From HPLC chromatogram, it shows that the compoundalmost pure (compound 1) and identified using NMR and MS.From MS data, compound 1 has molecular weight 408. Theanalysis of its NMR data and comparison with referenceshowed in Table 1. From the 13 C-NMR spectrum, compound 1indicated 25 carbons, including 8 sp 2 methine carbons, 4methylene carbons, 3 methyl groups, and 10 quartenerycarbons.Table 1. Comparison of 13 C and 1 H NMR assignment between compound 1and referenceFrom NMR spectrum, MS data, and comparison withprevious spectral data [16];[17];[18], compound 1 wasidentified as prenylated flavonoid, named 1-(2,4-dihydroxyphenyl)-3-[8-hydroxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1-benzopyran-5-yl]-1-propanone (Figure 3). In[18], this compound was isolated by general gravitationcolumn chromatography that needs various solid and mobilephase system for separations and purifications beforeobtaining the pure compound 1. Therefore this suggests theadvantage of using CCC system for isolation of compound.C No.13 C and 1 H NMR assignment fromcompound 1δ C (ppm)δ H (ppm)(H, multiplicity, J inHz)13 C and 1 H NMRassignment from reference[16]δ C (ppm)δ H (ppm)(mult., J inHz)1 165.20 165.152 103.62 6.38 (1H, d, 2.45) 103.51 6.37 (2.0)3 163.48 163.1446.37 (1H, dd, 2.45;6.34 (8.8;108.27 8.5) 107.97 2.0)5 132.45 7.54 (1H, d, 8.5) 132.27 7.55 (8.8)6 113.66 113.597 204.14 203.898 39.77 3.1 (2H, q) 39.65 3.08 (m)9 26.66 2.9 (2H, m) 26.49 2.97 (m)10 128.08 127.9111 121.29 6.60 (1H, d, 8.5) 121.14 6.61 (8.2)12 114.71 6.72 (1H, d, 8.5 ) 114.53 6.73 (8.2)13 143.11 143.0214 139.72 139.5615 119.22 119.0316 119.49 6.54 (1H, d, 9.8) 119.36 6.54 (10.2)17 130.30 5.63 (1H, d, 9.8) 130.12 5.63 (10.2)18 78.9 78.761940.77 1.70 (m)40.89 1.7 (2H, m)20 22.94 2.09 (2H, m) 22.80 2.08 (m)21 123.99 5.08 (1H, t) 123.84 5.08 (7.0)22 132.10 131.9523 25.80 1.66 (3H, s) 25.66 1.65 (bs)24 17.78 1.57 (3H, s) 17.64 1.56 (bs)25 26.20 1.38 (3H, s) 26.07 1.38 (s)1-OH 12.85 (1H, s) 12.81 (s)3-OH7.20 (bs)13-OH5.58 (bs)* Spectra compound 1 recorded at 500 MHz for 1 H spectrum and 125 MHzfor 13 C spectrum in CDCl 3 and reference’s spectra recorded at 500 MHz. Thevalues are in ppm and J values (Hz) in parentheses. Abbreviations for NMRsignal are as follows: s = singlet, d = doublet, and t = triplet.The 1 H-NMR spectra suggested the existence of two aromaticrings, ring A and B. At ring A have three proton aromatics,revealed a clear AMX system as indicated by the protonresonances at 6.38 (H-3’, d, J 2.45 Hz) and 6.37 ppm (H-5’,dd, J 2.45 Hz and 8.5 Hz) and at 7.54 ppm (H-6’, d, J 8.5 Hz)and 6.37 ppm (1 H, dd, J 2.45 Hz and 8.5 Hz). The down fieldchemical shift at 12.85 ppm (s) indicated the presence ofhydroxyl group formed a hydrogen bond with a carbonylgroup (C-1) at 204.14 ppm. The multiplicities of carbons wereassigned by the DEPT-135 experiment.HO32423OHA2216O716Fig. 3. Molecular Structure Compound Isolate A [1-(2,4-dihydroxyphenyl)-3-[8-hydroxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1- benzopyran-5-yl]-1-propanone].1910C181514IV. CONCLUSIONSCounter-current chromatography technique is moreefficient for compound isolation from ethyl acetate fraction ofA. altilis leaves compare to column chromatography because itdoesn’t need a lot of purification step.Based on NMR spectra, MS data, and compared withprevious published spectral data, compound 1 from ethylacetate extract of A. altilis leaves was identified as aprenylated flavonoid, [1-(2,4-dihydroxyphenyl)-3-[8-hydroxy-2-methyl-2-(4-methyl-3-pentenyl)-2H-1- benzopyran-5-yl]-1-propanone].ACKNOWLEDGMENTThis work is collaboration research between ZhejiangUniversity and Indonesian Institute of Sciences (LIPI). Wealso thanks to Prof. Yuanjiang Pan and his group member forfacilitating this research.BO2513OHJanuary 2013 ATBAS-30216065©Asian Transactions 8

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