Abstract
We have previously shown that ethanol or chloroform extracts of the leaves of Impatiens balsamina (LIB) have anti-tumor activity against the human hepatocellular carcinoma cell line HepG2. The ethanol extracts were separated into five fractions according to polarity. An MTT assay indicated that two of the fractions had anti-tumor activity and that the petroleum ether fraction (PEF) was the most active. But the available quantities of both the PEF and chloroform fractions (CHF) were limited, precluding further study. The chloroform extract (CHE) shared almost all the same spots with the PEF and CHF and was plentiful enough to carry out further separations. Thus, the CHE was further separated into six sub-fractions (CHE1~6) by column chromatography. A MTT assay showed that only the CHE2 fraction had a strong tumor inhibition ratio (IC50 = 6.47±0.05 mg/L), which was superior to that of curcumin (IC50 = 13.95±0.11 mg/L). However, TLC revealed that CHE2 was not pure and still contained two more components. After further separation and purification, followed by TLC and MTT assay confirmation, the final active component was isolated and identified as 2-methoxy-1,4-naphthoquinone by m.p., UV, MS and 13C- and 1H-NMR data. This is the first report demonstrating that 2-methoxy-1,4-naphthoquinone has intensive in vitro anti-tumor activity against HepG2 cells.
Highlights
The entirety of Impatiens balsamina plants have been widely used in Traditional Chinese Medicine to treat rheumatism, isthmus and crural aches, fractures, superficial infections, fingernail inflammation, etc. [1]
There were few common spots between those shared by the petroleum ether fraction (PEF), chloroform fractions (CHF) and chloroform extract (CHE) groups and those shared by the EAF, BUF, and WAF fractions
The results indicated that the PEF, CHF and CHE fractions all had some in vitro anti-tumor activity against HepG2 cells, with the PEF fraction possessing the strongest activity, having a cell inhibition ratio of approximately 80% at a concentration of 50 mg/L, but there were too many common spots among the three of them to determine which spot active was the source of the observed activity
Summary
The entirety of Impatiens balsamina plants (balsam, LIB) have been widely used in Traditional Chinese Medicine to treat rheumatism, isthmus and crural aches, fractures, superficial infections, fingernail inflammation, etc. [1]. Modern chemical and pharmacological studies have identified flavonol and naphthoquinone derivatives, some of which have strong antimicrobial [2], anti-anaphylaxis [3, 4], anti-inflammatory [5] as well as itch alleviating and anti-dermatitis activities, as the main chemical components of this plant [6]. A pilot study demonstrated that LIB has in vitro anti-tumor activity against the HepG2 human hepatocellular carcinoma cell line. We report the isolation of the active fractions using a MTT assay-guided fractionation and identified the positive spots using TLC. The in vitro anti-tumor activity of the active fractions was assessed. We purified and identified the active component of the fraction displaying the highest activity
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