Antioxidant and Phytochemical Potential of and Phytochemicals in Gymnema inodorum (Lour.) Decne in Northern Thailand.

Gymnema inodorum (Lour.) Decne is a vegetable local to Chiang Mai Province of Northern Thailand. This study aimed to analyze the antioxidant and phytochemical potential of G. inodorum found in Chiang Mai Province; antioxidant compounds of G. inodorum were tested via DPPH, ABTS and FRAP assays, and total phenolic compound and total flavonoid contents were analyzed. Anti-inflammatory effects were focused on regarding pharmacological potential. The gymnemic acid level was analyzed by HPLC-UV, and other potential chemicals were analyzed by LC-QTOF/MS. The quantifications of gymnemic acid contents analyzed using HPLC-UV showed that the highest gymnemic acid concentrations were found in the air-dried and roasted 1-day-fermented leaf extracts (0.1258 ± 0.0157 µg/mg). The highest free radical scavenging activity via DPPH assay was found in baked leaf extract, with an IC50 of 8.99 mg/mL, and via ABTS assay in baked and roasted leaf extracts, with an IC50 of 1.05 mg/mL. FRAP assays showed the highest free radical scavenging activity for the baked leaf extract, with 0.0085 ± 0.008 mM Fe2+/g sample. The total phenolic contents of fresh G. inodorum leaf extracts obtained with ethanol, methanol and water were 0.19 ± 0.0004, 0.21 ± 0.0010 and 0.10 ± 0.0008 µg GAE/g, respectively. The total flavonoid contents of fresh G. inodorum leaf extracts obtained with ethanol, methanol and water were 74.56 ± 28.00, 71.88 ± 16.11 and 10.74 ± 3.63 µg QE/g, respectively. The LC-QTOF/MS analysis of the fresh G. inodorum leaf extract showed that 6-hydroxykaempferol 7-rutinoside was the most abundant compound. In the study, G. inodorum, a plant local to Northern Thailand, is shown to be a useful plant with high antioxidant and phytochemical potential properties.

Tentative Identification of Phytochemicals from Smilax glabra and Smilax corbularia Extracts by LC-QTOF/MS and Their Bioactive Potential.

The Smilacaceae family has been used as a food source and herbal medicine for a long time. This study aims to identify the phytochemicals extracted from Smilax glabra and Smilax corbularia by using LC-QTOF/MS analysis and determine their bioactive potential. Compounds were identified from S. glabra and S. corbularia extracts by LC-QTOF-MS and it was found that longistylin A and CAY10435 have higher degrees of matching compounds (99.66% and 99.87%). Smilax glabra showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 71.94 ± 1.46% and 59.84 ± 4.80%, respectively, and FRAP at 730.69 ± 33.62 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.017 ± 0.001, 0.015 ± 0.001, and 0.016 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents were 0.043 ± 0.002, 0.033 ± 0.002, and 0.006 ± 0.003 mg QE/g, respectively. The anti-inflammatory capacity showed 97.26% protection and 2.74% hemolysis. The antimicrobial activity can inhibit Gram-positive bacteria with a minimum inhibitory concentration (MIC) of 62.5 mg/mL and a minimum bactericidal concentration (MBC) of 500 mg/mL. Smilax corbularia showed antioxidant capacity, i.e., DPPH and ABTS at percentage inhibitions of 72.24 ± 0.64% and 39.87 ± 2.37%, respectively, and FRAP at 208.33 ± 50.80 mg AAE/100 g sample. The total phenolic compound contents of the ethanol, methanol, and water extracts were 0.006 ± 0.000, 0.007 ± 0.002, and 0.002 ± 0.001 mg GAE/g, respectively, while the total flavonoid contents of the ethanol and methanol extracts were 0.012 ± 0.001 and 0.008 ± 0.000 mg QE/g, respectively. The anti-inflammatory capacity showed 96.64% protection and 3.36% hemolysis. The antimicrobial activity of the extracts can inhibit Gram-positive bacteria with a MIC of 31.25 mg/mL and MBC of 125 mg/mL for the ethanol extract and a MIC of 125 mg/mL and MBC of 62.5 mg/mL for the methanol extract. In conclusion, Smilax glabra and Smilax corbularia were found to contain several phytochemicals that can be used for further study. Both Smilax species can also be used as sources of antioxidants and herbal medicines for killing Gram-positive bacteria.

Recovery of Polyphenolic Fraction from Arabica Coffee Pulp and Its Antifungal Applications.

Coffee pulp is one of the most underutilised by-products from coffee processing. For coffee growers, disposing of this agro-industrial biomass has become one of the most difficult challenges. This study utilised this potential biomass as raw material for polyphenolic antifungal agents. First, the proportion of biomass was obtained from the Arabica green bean processing. The yield of by-products was recorded, and the high-potency biomass was serially extracted with organic solvents for the polyphenol fraction. Quantification of the polyphenols was performed by High Performance Liquid Chromatography (HPLC), then further confirmed by mass spectrometry modes of the liquid chromatography-quadrupole time-of-flight (QTOF). Then, the fraction was used to test antifungal activities against Alternaria brassicicola, Pestalotiopsis sp. and Paramyrothecium breviseta. The results illustrated that caffeic acid and epigallocatechin gallate represented in the polyphenol fraction actively inhibited these fungi with an inhibitory concentration (IC50) of 0.09, 0.31 and 0.14, respectively. This study is also the first report on the alternative use of natural biocontrol agent of P. breviseta, the pathogen causing leaf spot in the Arabica coffee.

Effect of Ethanolic Caesalpinia sappan Fraction on In Vitro Antiviral Activity against Porcine Reproductive and Respiratory Syndrome Virus.

Porcine reproductive and respiratory syndrome virus (PRRSV) is a major epidemic in pig production, leading to economic losses in the pig industry worldwide. The use of medicinal plants with antiviral properties might be useful help to prevent and control PRRSV outbreaks. Caesalpinia sappan (CS) heartwood is an important herbal ingredient used in Thai folk medicine, possessing various biological activities, including antiviral activity. The present study focuses on the in vitro antiviral activity against PRRSV of a semi-purified fraction of ethanolic CS crude extract using preparative high-performance liquid chromatography. Qualification of the fractions illustrating positive antiviral activity was carried out with liquid chromatography-quadrupole time-of-flight mass spectrometry. The preparative chromatography separated the crude extract into six consecutive fractions, among which the first fraction showed potential antiviral activity by inhibiting PRRSV replication in a MARC-145 monolayer (virus titer 2.75 median tissue culture infective dose (TCID50)/mL (log10) vs. 9.50 median log10 TCID50/mL of the control) at 72 h post-infection, and this fraction included byakangelicin, brazilin, naringenin, and brazilein. These results provide useful information for further study to effectively develop the CS bioactive antiviral compounds against PRRSV as a feed additive or veterinary drug in the pig industry.

The Pesticide Exposure of People Living in Agricultural Community, Northern Thailand.

BACKGROUND AND AIM: Biomarkers of pesticide exposure are generally lacking in communities where agricultural crops are grown. The purpose of this study was to focus on evaluating biomarkers of pesticide exposure in people living in an agricultural area of San Pa Tong District in Chiang Mai Province, northern Thailand. MATERIALS AND METHODS: One hundred and twenty-four participants (38 nonfarm workers, 38 rice growers, 31 longan growers, and 17 vegetable growers) from San Pa Tong District gave consent to participate in the study. Pesticide exposure was assessed by determining acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) levels in blood samples using Ellman's method and measuring 6-dialkylphosphate metabolites (DAPs), 3-phenoxybenzoic acid (3-PBA), and glyphosate in urine samples using chromatographic methods. RESULTS: AChE and BChE activities in the nonfarm worker group had higher level than those in the grower groups. DAPs were detected in almost all urine samples and 3-PBA was detected in 12-45% of each group, while glyphosate was found in 11 - 30% among the three groups of growers but not in nonfarm workers. CONCLUSION: In this study, participants living in an agricultural area of San Pa Tong District were exposed to organophosphate, synthetic pyrethroid, and glyphosate through multiple pathways.

Cross validation of gas chromatography-flame photometric detection and gas chromatography-mass spectrometry methods for measuring dialkylphosphate metabolites of organophosphate pesticides in human urine.

We report two analytical methods for the measurement of dialkylphosphate (DAP) metabolites of organophosphate pesticides in human urine. These methods were independently developed/modified and implemented in two separate laboratories and cross validated. The aim was to develop simple, cost effective, and reliable methods that could use available resources and sample matrices in Thailand and the United States. While several methods already exist, we found that direct application of these methods required modification of sample preparation and chromatographic conditions to render accurate, reliable data. The problems encountered with existing methods were attributable to urinary matrix interferences, and differences in the pH of urine samples and reagents used during the extraction and derivatization processes. Thus, we provide information on key parameters that require attention during method modification and execution that affect the ruggedness of the methods. The methods presented here employ gas chromatography (GC) coupled with either flame photometric detection (FPD) or electron impact ionization-mass spectrometry (EI-MS) with isotopic dilution quantification. The limits of detection were reported from 0.10ng/mL urine to 2.5ng/mL urine (for GC-FPD), while the limits of quantification were reported from 0.25ng/mL urine to 2.5ng/mL urine (for GC-MS), for all six common DAP metabolites (i.e., dimethylphosphate, dimethylthiophosphate, dimethyldithiophosphate, diethylphosphate, diethylthiophosphate, and diethyldithiophosphate). Each method showed a relative recovery range of 94-119% (for GC-FPD) and 92-103% (for GC-MS), and relative standard deviations (RSD) of less than 20%. Cross-validation was performed on the same set of urine samples (n=46) collected from pregnant women residing in the agricultural areas of northern Thailand. The results from split sample analysis from both laboratories agreed well for each metabolite, suggesting that each method can produce comparable data. In addition, results from analyses of specimens from the German External Quality Assessment Scheme (G-EQUAS) suggested that the GC-FPD method produced accurate results that can be reasonably compared to other studies.