Mitragyna KORTH. is a small genus in family Rubiaceae consisting of ten species in the world. Six species, M. speciosa (KORTH.) HAVIL., M. tubulosa (ARN.) HAVIL., M. parvifolia (ROXB.) KORTH., M. hirsuta HAVIL., M. diversifolia (WALL. ex G. DON) HAVIL., M. rotundifolia (ROXB.) O. KUNTZE, widely grow in Thailand, Vietnam, Philippines, Malay Peninsula, Sumatra, Borneo, and New Guinea islands. 1,2) In Thailand, M. hirsuta, M. diversifolia and M. rotundifolia are seen commonly. M. speciosa distributed from central to southern part, is a narcotic plant, so called "Kratom," but with specific medicinal importance.1,3) The leaves of M. speciosa have been chewed habitually by Thai natives for the opium-like effect and coca-like stimulant to overcome the burdens of their hard work. In Thai traditional medicine it has also been used to treat diarrhea, stop coughing and relieve injury pain, and in modern it has been used as a substitute for opium and for treatment of addiction to morphine.4) Phytochemical studies revealed that mitragynine, an indole alkaloid, was the major constituent of this plant, accounting for about half of the total alkaloid contents. Its analogues, speciogynine, speciociliatine, and paynanthine etc. were also found. 5,6) Recent pharmacological studies reported the antinociceptive effect of this species due to containing of mitragynine and its analogues. Especially, 7-hydroxymitragynine, a minor constituent, revealed significantly potent antinociceptive activity through opioid receptors.5-7) Its antinociceptive activity was many times more potent than morphine when administrated orally and subcutaneously. 8,9) The use of M. speciosa has been forbidden in Thailand due to its narcotic effects. The Thai government passed the law to make planting of M. speciosa illegal. However, since the species is indigenous to Thailand and widely distributed, the law is hard to raise its efficiency. On the other hand, the leaves of M. diversifolia or others are frequently used as substitutes but are not considered as effective. Therefore, authentication of M. speciosa is essential for both forensic and medicinal purpose.In recent years, molecular identification of herbal drugs by nucleotide sequence of various DNA regions has been demonstrated to be a powerful way.10-16) Especially, DNA regions with high evolutionary rate have been widely used to discriminate species and investigate phylogenetic relationship among closely-related taxa, such as chloroplast trnK/matK, 11) rpl14-16, 12) trnT-F, 13) and nuclear internal transcribed spacers (ITS), [13][14][15] 5S spacer region, 16) etc. In a preliminary study, we found that there was no nucleotide difference in chloroplast matK gene and nuclear 18S rRNA gene regions between M. speciosa and M. hirsuta (unpublished data). In the present study, we investigated the sequence of ITS region for characterizing four Mitragyna species in Thailand and further to discriminate M. speciosa from the other species. MATERIALS AND METHODS Plant MaterialsTotal of seventeen specimens of four Mi...
As a continuous searching for anti-diabetic(type II) substances, seven mucilage polysaccharides from selected plants were studied as follow: aerial parts of Basella alba Linn., fruits of Hibiscus esculentus Linn., leaves of Litsea glutinosa (Lour.) C.B. Robinson, seeds of Ocimum canum Sims., seeds of Plantago ovata Forssk., fruits of Scaphium scaphigerum G. Don. and seeds of Trigonella foenum-graecum Linn. The bioactive properties for entrapping glucose, inhibiting enzyme alpha-glucosidase and free radical scavenger were in vitro studied compared to glucomannan. The physical characteristics for water holding capacity and viscosity were determined. The chemical characteristics were assayed for monosaccharide composition using methanolysis, TMSderivatization and gas chromatography. O. canum mucilage superiorly entrapped glucose compared to glucomannan. This activity was relevant to its highly viscous gelation. S. scaphigerum showed another property of alphaglucosidase inhibition. S. scaphigerum mucilage (0.5%) inhibited the enzyme activity by 82.6%, compared to 1-Deoxynorjirimycin (by 47.6%). Most mucilages, except O. canum and P. ovata, showed DPPH scavenging activity higher than glucomannan. Galacturonic acid was found in 3 from 7 mucilages namely B. alba, P. ovata and S. scaphigerum. Whereas rhamnose was common sugar found in all seven mucilages. Monosaccharide components of these mucilages were compared to the results from the previous reports. Sci
Objective: To develop a thin-layer chromatography (TLC) densitometric method for the determination of oxyresveratrol content in Artocarpuslakoocha heartwood and in the traditional drug ‘Puag-Haad’. Materials and Methods: Sample solution of A. lakoocha heartwood was prepared by Soxhlet extraction of the plant material in ethanol, whereas the Puag-Haad solution was obtained by dissolving the drug in methanol. Analysis of each sample solution was performed on a Silica gel 60 F254 TLC plate (20 × 10 cm) with methylene chloride/methanol (85:15) as the mobile phase. After development, the TLC plate was examined with a TLC scanner in the absorbance mode at 254 nm. The newly developed analytical method was validated using an authentic sample of oxyresveratrol previously isolated from A. lakoocha heartwood, and was used to analyze the oxyresveratrol content in samples of A. lakoocha heartwood and the traditional drug Puag-Haad. Results: A sensitive and reliable TLC densitometric method was successfully developed. The method was validated in terms of accuracy (99.11–102.60%) and precision (1.66–4.23% coefficient of variation). The limits of detection and quantitation were 15.6 and 52 ng/spot, respectively. The amounts of oxyresveratrol in 3 samples of A. lakoocha heartwood collected from its natural habitat were 49.0–182.3 mg/g, whereas those in 11 commercial samples were in the range of 23.4–69.6 mg/g. The oxyresveratrol contents in 2 samples of traditional drug Puag-Haad were 780.1 and 837.5 mg/g. Conclusion: The TLC densitometric method developed in this study is a simple, convenient, sensitive and reliable procedure. It was an effective analytical tool for the evaluation of oxyresveratrol content in both A. lakoocha heartwood and the traditional drug Puag-Haad.
The chlorogenic acid, rosmarinic acid, and caffeic acid contents in 100 selected plants were determined using reversed phase high performance liquid chromatography equipped with diode array detector. The optimum condition was 0.2% phosphoric acid in water (solvent A) and methanol (solvent B) as the mobile phase, which was set at 45% B for 20 minutes at a flow rate of 1.2 mL/min. The column temperature was maintained at 30 °C and the detection wavelength was 325 nm. Among 100 selected plants, 39.64% contained all 3 compounds, 40.54% contained 2 compounds, 14.41% contained only 1 compound, and 5.41% could not detect any of the 3 compounds. The highest contents of chlorogenic acid, rosmarinic acid, and caffeic acid were found in Lonicera japonica flowering buds, Melissa officinalis leaves, and Coffea canephora seeds at the concentration of 9.900 ± 0.004, 19.908 ± 0.171, and 1.233 ± 0.003 g/100 g of dried plant, respectively.
Simple equations and theoretical models, related to enantioselectivity (kappa) and C, have been developed for prediction of electrophoretic mobility difference (Deltamu) and separation selectivity (alpha) for enantiomers in CE using dual CDs, where alpha and kappa are defined as the ratio of mu and the ratio of binding constant (K) for enantiomers to each CD, respectively, C the CD concentration, and the average K for enantiomers and each CD. Experiments were carried out using dual CDs as beta-CD and dimethyl-beta-cyclodextrin (DM-beta-CD) and test analytes as five pairs of amphetamine drug enantiomers. A change in observed Deltamu and alpha of enantiomers in dual CDs was found to be in excellent agreement with the theoretical models. For example, in comparison with single CD1, dual CDs can enhance Deltamu and alpha up to the maximum value when enantiomers migrate with the same order in CD1 and CD2, and have the value of rho > 1.0, where rho is the enantioselectivity ratio for CD2 to CD1, while worse Deltamu and alpha are obtained for enantiomers with rho < 1.0.
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