Wandee Gritsanapan

Comparison of bioactive compounds content, free radical scavenging and anti-acne inducing bacteria activities of extracts from the mangosteen fruit rind at two stages of maturity.

Contents of bioactive components, free radical scavenging and anti-acne producing bacteria activities of young and mature fruit rind extracts of mangosteen were compared. The young fruit rind extract contained significantly higher contents of phenolics and tannins and promoted higher free radical scavenging activity than the mature fruit rind extract, while the later extract contained higher contents of flavonoids and alpha-mangostin xanthone and gave higher anti-acne producing bacteria activity than the young fruit rind extract. Thus, the young and mature stages of mangosteen fruit rind should be beneficial for further development of antioxidant and anti-acne pharmaceutical preparations, respectively.

Determination of Berberine Content in the Stem Extracts of Coscinium fenestratum by TLC Densitometry

Objective: To develop the optimal extraction procedure (i.e. maceration, percolation or Soxhlet extraction) and thin- layer chromatographic (TLC)-densitometric method for the determination of berberine content of Coscinium fenestratum. Materials and Methods: Maceration, percolation and Soxhlet extraction techniques were used to extract alkaloids from dried stems of C. fenestratum. The solvents used were 50 and 80% ethanol. Crude extracts and berberine content recovered from the TLC fingerprint were evaluated for chemical components of each extraction method. Precoated silica gel GF254 plates were used as stationary phase while butanol:glacial acetic acid:water (14:3:4) was used as a mobile phase. Detection and quantitation of berberine were performed by densitometry at the wavelength of 415 nm over the linearity range of 240–840 ng (r2 = 0.9982). The relative standard deviations from intraday and interday precisions were less than 4.13%. Results: The recovery of standard berberine was 97.58–98.71% (%RSD = 3.85), and the limit of detection and quantitation were 25 and 50 ng/spot, respectively. Eighty percent ethanol gave a higher content of berberine than 50% ethanol. Berberine contents from maceration, percolation and Soxhlet extraction with 80% ethanol were 3.37± 0.30, 3.08± 0.38 and 2.67± 0.27% w/w, respectively. Conclusion: The TLC-densitometric method was simple, accurate and precise for quantitating berberine in the stem extract of C. fenestratum. Maceration with 80% ethanol gave the highest content of berberine in the extract. TLC of the extracts from different methods showed a similar pattern.

Phytochemical and antioxidant studies of Laurera benguelensis growing in Thailand

The aim of this study was to investigate metabolites of the lichen Laurera benguelensis. A high-performance liquid chromatographic (HPLC) method has been developed for the characterization of xanthones and anthraquinones in extracts of this lichen. Lichexanthone, secalonic acid D, norlichexanthon, parietin, emodin, teloschistin and citreorosein were detected in the lichen samples, which were collected from two places in Thailand. Components of the lichen were identified by relative retention time and spectral data. This is the first time that a detailed phytochemical analysis of the lichen L. benguelensis was reported and this paper has chemotaxonomic significance because very little has been published on the secondary metabolites present in Laurera species. Some of the metabolites were detected for the first time in the family Trypetheliaceae. The results of preliminary testing of benzene extract and its chloroform and methanol fractions showed that all samples showed a weak radical scavenging activity. The chloroform extract showed the highest antioxidant activity.

Damnacanthal, a noni component, exhibits antitumorigenic activity in human colorectal cancer cells

Damnacanthal, an anthraquinone compound, is isolated from the roots of Morinda citrifolia L. (noni), which has been used for traditional therapy in several chronic diseases including cancer. Although noni has been consumed for a long time in Asian and Polynesian countries, the molecular mechanisms by which it exerts several benefits are starting to emerge. In this report, we examined systematic approaches on the cancer-suppressing capability of damnacanthal in colorectal tumorigenesis. Damnacanthal exhibits cell growth arrest as well as caspase activity induction in colorectal cancer cells. We also examined several potential target proteins and found that the proapoptotic protein nonsteroidal anti-inflammatory activated gene-1 (NAG-1) is highly induced. Subsequently, we have found that damnacanthal also enhances transcription factor CCAAT/enhancer binding protein β (C/EBPβ), which controls NAG-1 transcriptional activity. Blocking of C/EBPβ by shRNA results in the reduction of NAG-1 expression as well as caspase activity in the presence of damnacanthal. Taken together, these results indicate that damnacanthal increases antitumorigenic activity in human colorectal cancer cells and that C/EBPβ plays a role in damnacanthal-induced NAG-1 expression.

Analysis of naphthoquinone derivatives in the Asian medicinal plant Eleutherine americana by RP-HPLC and LC–MS

The first analytical procedure for the determination of a new naphthopyrone, eleutherinoside A, together with the known bioactive compounds eleuthoside B, isoeleutherin, eleutherin and eleutherol in Eleutherine americana was established. Optimum HPLC separation of these naphthoquinone derivatives was possible on RP-12 column material, using water and acetonitrile as mobile phase. Flow-rate, detection wavelength and temperature were adjusted to 1.0 mL/min, 254 nm and 40 degrees C, respectively. Validation results indicated that the HPLC method is well suited for the determination of naphthoquinone derivatives in the bulbs of E. americana with a good linearity (r2>0.9996), precision (intra-day R.S.D. <4.70%, inter-day R.S.D. <5.68%) and recovery rates from 96.26 to 103.48%. Limit of detection (LOD) was found to be below 0.84 microg/mL for all five compounds. LC-MS analyses performed in positive and negative electrospray ionization mode assured peak purity and identity. The analysis of different E. americana samples from Thailand revealed that eleutherol (0.10-0.20%) was dominant in all specimens, followed by isoeleutherin and eleutherin. The new natural product 2,5-dimethyl-10-hydroxynaphthopyrone 8-O-beta-d-glucopyranoside occurred in percentages of less than 0.05%.

Free radical scavenging and anti-acne activities of mangosteen fruit rind extracts prepared by different extraction methods.

The ethanol extracts of mangosteen fruit rinds prepared by several extraction methods were examined for their contents of bioactive compounds, DPPH-scavenging activity, and anti-acne producing bacteria against Propionibacterium acnes and Staphylococcus epidermidis. The dried powder of the fruit rind was extracted with 95% ethanol by maceration, percolation, Soxhlet extraction, ultrasonic extraction, and extraction using a magnetic stirrer. Soxhlet extraction promoted the maximum contents of crude extract (26.60% dry weight) and α-mangostin (13.51%, w/w of crude extract), and also gave the highest anti-acne activity with MIC 7.81 and 15.63 μg/mL and MBC 15.53 and 31.25 μg/mL against P. acnes and S. epidermidis, respectively. Ethanol 70% and 50% (v/v) were also compared in Soxhlet extraction. Ethanol 50% promoted the extract with maximum amounts of total phenolic compounds (26.96 g gallic acid equivalents/100 g extract) and total tannins (46.83 g tannic acid equivalents/100 g extract), and also exhibited the most effective DPPH-scavenging activity (EC50 12.84 μg/mL). Considering various factors involved in the process, Soxhlet extraction carried a low cost in terms of reagents and extraction time. It appears to be the recommended extraction method for mangosteen fruit rind. Ethanol 50% should be the appropriate solvent for extracting free radical-scavenging components, phenolic compounds, and tannins, while 95% ethanol is recommended for extraction of α-mangostin, a major anti-acne component from this plant.

Structural relationships of stemona alkaloids: assessment of species-specific accumulation trends for exploiting their biological activities.

On the basis of a comparison of 42 Stemona samples, representing eight different species collected and cultivated in Thailand, species-specific accumulation trends of Stemona alkaloids were analyzed. An overview was achieved by comparative HPLC analyses of methanolic crude extracts of underground parts coupled with diode array or evaporative light scattering detectors. All major compounds were isolated and their structures elucidated by NMR and MS analyses. Protostemonine- and stichoneurine-type derivatives dominated, from which the latter characterize S. tuberosa and S. phyllantha accumulating species-specific isomers of tuberostemonine (3). The widespread S. curtisii and S. collinsiae clearly deviate by protostemonine-type derivatives dominated by stemofoline (10) and/or didehydrostemofoline (11). Further diversification within this structural type results from a mutual accumulation of derivatives with a pyrrolo- or pyridoazepine nucleus, leading to chemical variability in S. curtisii and S. aphylla.

Anti-Acne-Inducing Bacterial Activity of Mangosteen Fruit Rind Extracts

Objective: The aims of this study were to determine the most effective solvent extract of mangosteen, anti-acne- inducing bacterial activity and the amount of α-mangostin, a major active component in each mangosteen fruit rind extract, using high-performance liquid chromatography (HPLC). Materials and Methods: The fruit rinds of mangosteen were extracted with hexane, dichloromethane, ethanol and water. The extracts were tested for antibacterial activity against bacteria that induce acne, including Propionibacterium acnes and Staphylococcus epidermidis. Thin-layer chromatographic autobiography against these bacteria was also performed for each extract, while the α-mangostin content was analyzed using a validated HPLC method. Results: The dichloromethane extract exhibited the strongest antibacterial effect with minimum inhibitory concentration values for both bacterial species at 3.91 µg/ml, while the minimum bactericidal concentration values against P. acnes and S. epidermidis were 3.91 and 15.63 µg/ml, respectively. Thin-layer chromatographic autobiography indicated that α-mangostin was present in all extracts, except the water extract, and is a major active component against both P. acnes and S. epidermidis. Using HPLC, the dichloromethane extract yielded the highest content (46.21% w/w) of α-mangostin followed by the ethanol extract (18.03% w/w), the hexane extract (17.21% w/w) and the water extract (0.54% w/w). Conclusions: Dichloromethane extract exhibited the strongest anti-acne-inducing bacterial effect and this extract yielded the highest amount of α-mangostin.

Effect of Siam weed extract and its bioactive component scutellarein tetramethyl ether on anti-inflammatory activity through NF-κB pathway.

ETHNOPHARMACOLOGICAL RELEVANCE Siam weed (Chromolaena odorata (L.) King and Robinson) is a medicinal herb used for wound healing and inflammation-related diseases. AIM OF THE STUDY In this study, we evaluated the molecular mechanism by which Siam weed extract (SWE) and its bioactive components, scutellarein tetramethyl ether (scu), stigmasterol, and isosakuranetin affect anti-inflammatory activity. MATERIALS AND METHODS The expression of several inflammatory proteins in RAW 264.7 (murine) macrophages was assessed by Western blot and reverse transcription-polymerase chain reaction (RT-PCR). Biochemical assays including prostaglandin E2 (PGE2) and nitric-oxide (NO) quantification were performed. Luciferase promoter activity and immunocytochemistry of Nuclear factor-κB (NF-κB) were investigated. RESULTS Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are critical pro-inflammatory proteins. The level of protein and mRNA expression of these enzymes induced by lipopolysaccharide (LPS) was dramatically suppressed by treatment with SWE, scu, or stigmasterol compounds in a dose-dependent manner. They also reduced PGE2 and NO release. We further analyzed the NF-κB pathway and found that the scu compound suppressed IκB kinase complex alpha/beta (IKKα/β) and Inhibitory-kappa-B-alpha (IκBα), thereby suppressing COX-2 and iNOS expression. CONCLUSION This is the first report of the anti-inflammatory molecular mechanism in SWE and/or its bioactive component scu, indicating alteration NF-κB pathway and further providing potential uses in the treatment of inflammatory-related diseases.

In vitro antigonococcal activity of Coscinium fenestratum stem extract

ETHNOPHARMACOLOGICAL RELEVANCE Gonorrhea is a sexually transmitted disease (STD), which originates from bacteria, Neisseria gonorrhoeae. It is still one of the major health problems worldwide in both men and women. Many medicinal plants have been recorded in Thai folk medicine for relieving STD but there is no scientific report of these plants for antigonococcal activity. AIM OF THE STUDY This study was conducted to evaluate antigonococcal activity of 22 Thai medicinal plants selected from the plants popularly used in Thai folk medicine for treatment of gonorrhea. MATERIALS AND METHODS This study was performed by disc diffusion and agar dilution methods against Neisseria gonorrhoeae. Active compound was investigated by bioautographic assay. RESULTS Among the selected plants, Coscinium fenestratum (Gaertn.) Colebr. extract showed the most effective activity against Neisseria gonorrhoeae ATCC 49226 with MIC value of 47.39 microg/ml. Bioautographic assay revealed that berberine was the active compound of Coscinium fenestratum against Neisseria gonorrhoeae. The average MIC values of purified berberine against Neisseria gonorrhoeae ATCC 49226 and 11 clinical isolates were 13.51 and 17.66 microg/ml, respectively while average MIC value of the crude extract of Coscinium fenestratum against all clinical isolates was about 56.39 microg/ml. There was no acute toxicity detected at the dose of 5g of Coscinium fenestratum crude extract per kilogram. CONCLUSIONS These results provide theoretical support for ethnopharmacological relevance of antigonococcal activity of Coscinium fenestratum and its active compound.