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Introduction: Cyanthillium cinereum has long been used in Thai traditional medicine. However, the standard information of C. cinereum raw material has not been established. Objective: This study aimed to present the standard information of C. cinereum raw material. Material and methods: The microscopic and macroscopic characteristics, the physicochemical analysis, and assays for bioactive contents of 24 samples of C. cinereum from different locations were evaluated to determine the standard information of the plant material. Results: The result showed the microscopic characteristic of pollens in all powder samples. No nicotine content in C. cinereum extract was observed with LC-MS; apigenin and luteolin were instead selected as markers. The observed average contents of apigenin and luteolin were 0.238 and 1.059 mg/g, respectively, using HPLC-UV. Nitrite and nitrate components in C. cinereum, which induced flavor aversion leading to smoking cessation, were determined. The average nitrite and nitrate content were 0.004 and 0.526 mg/kg in ethanol extract and 0.014 and 1.645 mg/kg in the water extract, respectively. The results did not substantiate the concern for high potassium content in C. cinereum materials for cardiovascular and renal patients where only traces were observed in all samples (3.113 and 20.898 mg/kg in ethanol and water extracts, respectively). Conclusion: The results can be used to establish the specification of C. cinereum, which will be useful for quality control of the raw material. This new information is crucial and can be utilized extensively prior to herbal product preparation.

Asian rice, or Oryza sativa L., is a material for many diets. Rice hull is mostly indigestible to humans because of its enriched fiber components. The rice is commonly sold as white rice, which the brown paddy rice is polished. The pericarp, seed coat, nucellus, and aleurone layer that polished off is called rice bran, wherein the oil obtained from bran is called rice bran oil (RBO). RBO has gained popularity as a heart-healthy oil since it is a good source of valuable nutrients including gamma oryzanol, vitamin E, and phytosterols that exhibit antioxidant and hypocholesterolemic effects. Cold pressed extraction is a green alternative method for producing RBO, which is suited to a small-sized enterprise. Several studies are being performed around the world for the use of RBO and its by-products in health supplements, cosmetics, and the food industry. This chapter reviews the production, bioactive compounds, functionality, and applications of RBO.

Recently, a traditional Thai antihypertensive herbal recipe has reportedly been used in Thailand. Its ingredients have long featured in traditional Thai medicine preparations; however, research indicates that the presence of one of them - Tinospora crispa - may have negative effects on the liver and kidneys. Thus, the safety data of this recipe must be proved in animal models prior to conducting any studies in humans. The present case study aims to evaluate the safety of this recipe in Swiss albino mice and Wistar rats through acute and sub-chronic toxicity studies, respectively. The quality control of this recipe was also achieved to guarantee the chemical consistency throughout the entire experiment. Results showed that this recipe did not cause death or any toxic signs in mice or rats. The oral LD50 value in mice was more than 5.0 g/kg. Some hematological and serum biochemical values of treated rats, such as hematocrit, hemoglobin, platelet, monocytes, aspartate aminotransferase, bilirubin, and creatinine, were found to be statistically different from the control group; however, all values were within the ranges of normal rats. Considering the histological study, no damage on liver and kidney tissues was observed in the treatment.

Despite the fact that many researchers have developed different methods for the determination and detection of bile acid sequestrant, the analytical instruments are still quite costly, and expertise is usually required. In order to avoid the above-mentioned problems, simple, rapid, effective, and inexpensive methods to evaluate bile acid-binding are essential. The purpose of this study was to develop a simple and rapid thin-layer chromatography (TLC)—image analysis method to quantitatively determine the bile acid-binding activity of ten edible ethanolic plant extracts. The method was carried out by using TLC silica gel 60 F254 plate with the mobile phase composed of chloroform, methanol, and acetic acid (7:2:1 v/v). The chromatograms were analyzed with image analysis software (ImageJ). The method provided good linearity, accuracy, reproducibility, and selectivity for the determination of bile acid-binding activity. Litsea petiolata extract was found to show the most potent bile acid-binding activity compa...

The health benefits of herbs and herbal products are gaining more attention in southeast Asia. The World Health Organization (WHO) has been supporting countries to promote application of traditional medicines so that this valuable resource is utilized safely and effectively. In Thailand, many traditional herbal recipes have been established since ancient times. Since then, they have been carefully modified, based on the wisdom of traditional Thai medicine practitioners. For this study, a traditional Thai antihypertensive herbal recipe (TTAH) was selected and studied in detail. According to WHO guidelines, both analysis of a sizeable chemical constituent, and formulation data of a product, are a requirement to support a clinical trial for an herbal recipe. Therefore, high-performance liquid chromatography–mass spectrometry (LC–MS) was used to investigate the chemical fingerprints, chemical constituents, and putative active ingredients of the TTAH. Eight chemical fingerprints were established. Metabolic profiling of 10 possible compounds was also identified and all were shown to be active pharmaceutical compounds. An attempt was also made to prepare a suitable formulation of the TTAH, to standardize the amount of active ingredients per dose, and to improve patient compliance. All evaluated parameters guided us to prepare the TTAH as a capsule. This informative data can be included in part of the chemistry–manufacturing–control guidance prior to phase 1/2 clinical trials.

Bioactive peptides from natural sources are utilized as food supplements for disease prevention and are increasingly becoming targets for drug discovery due to their specificity, efficacy and the absence of undesirable side effects, among others. Hence, the ‘SpirPep’ platform was developed to facilitate the in silico-based bioactive peptide discovery of these highly sought-after biomolecules from Spirulina(Arthrospira platensis) and to select the protease (thermolysin) used for in vitro digestion. Analysis of the predicted and experimentally-derived peptides suggested that they were mainly involved in ACE inhibition; thus, an ACEi assay was used to study the ACE inhibitory activity of five candidate peptides (SpirPep1-5), chosen from common peptides with multifunctional bioactivity and 100% bioactive peptide coverage, originating from phycobiliproteins. Results showed that SpirPep1 inhibited the activity of ACE with IC50 of 1.748 mM and was non-toxic to fibroblasts of African green monkey kidney and human dermal skin. The molecular docking and MD simulation analysis revealed SpirPep1 had significantly lower binding scores than others and showed greater specificity to ACE. The non-bonded interaction energy of SpirPep1 and ACE was −883 kJ/mol. The SpirPep1 indirectly bound to ACE via the ACE substrate binding sites residues (D121, E123, S516, and S517) found in natural ACE inhibitory peptides (angiotensin II and bradykinin potentiating peptides). In addition, two unreported substrate binding sites including R124 and S219 were found. These results indicate that ‘SpirPep’ platform could increase the success rate for natural bioactive peptide discovery.

Tryptophan decarboxylase (TDC) catalyzes the decarboxylation of tryptophan to tryptamine in mitragynine biosynthesis via the shikimate pathway. Using the rapid amplification of cDNA ends (RACE) technique, the gene encoding TDC from Mitragyna speciosa was cloned (designated as MsTDC). The MsTDC cDNA contained an open reading frame (ORF) of 1,521 base pairs (bp) encoding 506 amino acid residues. It had a pyridoxal-phosphate (PLP)-binding site at the amino acid position 313–334 residues. The MsTDC showed homology of 68–76 % to the TDC of other plants. Heterologous expression in Escherichia coli afforded the soluble proteins as an apparent band of 57 kDa as judged by SDS-PAGE. Expression of the MsTDC in M. speciosa hairy roots under the 35S promoter was performed by insertion of MsTDC into pCAMBIA1300-gfp. The transgenic hairy root lines were detected by fluorescence microscopy and showed an increased accumulation of tryptamine.

Mitragynine is a pharmacologically-active terpenoid indole alkaloid found in Mitragyna speciosa leaves. Treatment with methyl jasmonate (10 μM) for 24 h and yeast extract (0.1 mg/ml) for 12 h were the optimum conditions of elicitation of mitragynine accumulation in a M. speciosa shoot culture. The former elicitor gave 0.11 mg mitragynine/g dry wt. Tryptophan decarboxylase and strictosidine synthase mRNA levels were enhanced in accordance with mitragynine accumulation.

1-Deoxy- d -xylulose 5-phosphate synthase (DXS, EC: 4.1.3.37), the first enzyme in the 2 C -methyl- d -erythritol 4-phosphate (MEP) pathway, is known to be responsible for the rate-limiting step of isoprenoid biosynthesis in Escherichia coli and Arabidopsis thaliana . In this study, the dxs gene from Croton stellatopilosus , designated csdxs , was cloned from leaf tissue using the rapid amplification of cDNA ends (RACE) technique. Leaves of C. stellatopilosus contain plaunotol, an acyclic diterpene alcohol. The csdxs cDNA containing the open reading frame of 2163 base pairs appeared to encode a polypeptide of 720 amino acids. Analysis of the deduced amino acid sequence revealed that the NH 2 -terminus of CSDXS carried a chloroplast transit peptide, a thiamine diphosphate binding site, and a transketolase motif, which are the important characteristics of DXS enzymes in higher plants. Multiple alignments of CSDXS with other plant DXSs have indicated that CSDXS has identity ranging between 68% and 89%. Expression levels of csdxs and genes encoding key enzymes in the plaunotol biosynthetic pathway, namely 2 C -methyl- d -erythritol 4-phosphate synthase ( meps ) and geranylgeranyl diphosphate synthase ( ggpps ), were analysed by measuring transcript levels in leaves of different developmental stages. The results showed that dxs , meps, and ggpps are all active in young leaves prior to full expansion when plaunotol is synthesised from the DXP precursor in chloroplasts. The dense presence of chloroplasts and oil globules in the palisade cells of these leaves support the view that these genes are involved in plaunotol biosynthesis in chloroplast-containing tissues.

Background: Fifteen medicinal plants used in traditional Thai herbal medicine (HM) for hypertension treatment were previously tested for their inhibitory activity against an angiotensin-converting enzyme (ACE). The ethyl acetate extract from one of these medicinal plants, Senna garrettiana, possessed satisfactory ACE inhibitory (ACEi) activity. Objective: For this study, S. garrettiana extract was further subjected to an isolation process to uncover the active compounds with ACEi activity. Materials and Methods: The ethyl acetate fraction was subjected to isolate and purify by column chromatography accompany with the ACEi assay. The structures of all compounds were elucidated using nuclear magnetic resonance spectroscopy. Results: It was found that S. garrettiana extract possessed two active compounds, piceatannol and betulinic acid, which both had good ACEi activity displaying IC50values of 8.44 μM and 26.77 μM, respectively. Conclusions: These findings show that both compounds can be used as markers for quality control of any standardized HM for hypertension treatment and not just for traditional Thai herbal remedies. Moreover, to the best of our knowledge, this is the first report investigating both compounds for their ACEi activity. Abbreviations used: ACE: Angiotensin-converting enzyme; ACEi: Angiotensin-converting enzyme inhibitory; CH2Cl2: Dichloromethane; DMSO: Dimethyl sulfoxide; HA: Hippuric acid; HCl: Hydrochloric acid; HHL: Hippuryl-L-histidyl-L-leucine; HM: Herbal medicine; HMs: Herbal medicines; HPLC: High-performance liquid chromatography; IC50: The half maximal inhibitory concentration; L-NAME: N (ω)-nitro-L-arginine methyl ester; MeOH: Methanol; NaCl: Sodium chloride; NADPH: Nicotinamide adenine dinucleotide phosphate; NMR: Nuclear magnetic resonance spectroscopy; NO: Nitric oxide; TDR: The Special Programme for Research and Training in Tropical Disease; μL: Microliter; μM: Micromolar.

Rhinacanthins-rich Rhinacanthus nasutus (RRn) extract was prepared and standardized to contain total rhinacantins not less than 70% w/w. In this study, antimicrobial activities of the RRn extract as well as rhinacanthin-C against Streptococcus mutans, Propionibacterium acnes, Helicobacter pylori, Staphylococcus aureus, S. epidermidis and Candida albicans were evaluated by microdilution assay. It was found that the RRn extract exhibited potent bactericidal activity against S. mutans with MIC and MBC of 4 microg/ml, and potent bacteriostatic activity against S. epidermidis, P. acnes and S. aureus with the MICs of 8-16 microg/ml. However, the RRn extract was not active against C. albicans at concentration up to 2000 microg/ml. The antimicrobial activities of the RRn extract was almost equal to those of rhinacanthin-C. Stability evaluations of the RRn extract through a period of 4 months found that the RRn extracts were stable when kept in a well-closed container protected from light and stored either under 4+/-2 degrees C or 30+/-2 degrees C. The RRn extracts that were exposed to light were not stable after 1 week of storage. Under accelerated conditions at 45 degrees C with 75% relative humidity, the RRn extracts were not stable after 8 weeks of storage. Study on the effect of pH on the stability of the aqueous methanolic solution of the RRn extract found that the solutions were not stable at pH 5.5, pH 7.0, and pH 8.0. However, the rhinacanthin solution at pH 5.5 was more stable than those at pH 7.0 and pH 8.0.

A newly developed liquid chromatography-mass spectrometry (LC-MS) method for the analysis of cold pressed rice bran oil (RBO) was established and used to discriminate between RBOs produced from two different cultivars of major Thai fragrant rice species. The cold pressed RBO was prepared using the screw compression method. The LC-MS data were preprocessed with MZmine 2.10 program before evaluating with principal component analysis using SIMCA 13 software. The LC-MS method was able to detect and quantify several kinds of valuable constituents such as fatty acids, vitamin E, and γ-oryzanol. The chromatographic condition was feasible; short time for analysis and simple method were achieved. From score plot and loading plot of principle component analysis (PCA) , two rice cultivar samples were clearly separated, and it was revealed that Khao-Hom-Pathum was more suitable than Khao-Hom-Mali for cold pressed RBO production since it contained high total γ-oryzanol and less saturated free fatty acids. As with the fixed price of all the rice brans, this information can be used in order to, if possible, preserve the price of rice brans from different cultivars.

A reversed-phase high-performance liquid chromatographic method is described for the simultaneous determination of rhinacanthin-C, rhinacanthin-D, and rhinacanthin-N in Rhinacanthus nasutus leaves. The method involved the use of a TSK-gel ODS-80Ts column (5 microm, 4.6 x 150 mm i.d.) with the mixture of methanol and 5% aqueous acetic acid (80:20, v/v) as the mobile phase. The parameters of linearity, repeatability, accuracy, and specificity of the method were evaluated. The recovery of the method was 94.3-100.9%, and good linearity (r(2) > or = 0.9999) was obtained for all rhinacanthins. A high degree of specificity as well as repeatability and reproducibility (relative standard deviation values less than 5%) were also achieved. The limit of detection and quantification of all rhinacanthins were 0.75 and 3.0 microg/mL, respectively. The solvents for extraction of rhinacanthins from R. nasutus leaves were examined in order to obtain the leaf extract with high rhinacanthin content. It was found that ethyl acetate was an appropriate solvent for rhinacanthin extraction. Fractionation of the ethyl acetate extract using a basic anion exchange resin (Amberlite IRA-67) eluted with 10% acetic acid in methanol afforded a rhinacanthin-rich extract (HRn). The total content of rhinacanthins was increased from 37.4% w/w to 77.5% w/w. The antifungal activities of HRn against Trichophyton rubrum, T. mentagrophytes, and Microsporum gypseum were also improved.