Standardization of Leonurus sibiricus L. aerial part and capillary electrophoresis quantitative analysis of its leonurine content.

The quality parameters of Leonurus sibiricus L. aerial part crude drugs were evaluated. Fifteen crude drugs were collected from various locations throughout Thailand. The transverse section of the stem of L. sibiricus showed quadrangular character highlighted the ribs with angular collenchyma. The epidermis was uniseriate with abundant glandular trichomes distribution. Prismatic calcium oxalate prisms were found in the stem medullary parenchyma.The histological character of crude drug powder showed bordered pitted vessel, fragment of fiber, glandular trichome, prism crystal, spiral vessel, starch granule, and stomata. The loss on drying, total ash, acid-insoluble ash, and moisture contents should be not more than 8.18, 15.28, 4.04, and 8.91 g/100 g dry weight, whereas ethanol and water-soluble extractive values should be not less than 7.67, and 17.21 g/100 g of dry weight, respectively. Leonurine in the crude drugs were analyzed by capillary electrophoresis (CE) with photodiode array detector. The ethanolic extraction performed by Soxhlet apparatus yielded 18.86 ± 4.09 g/100 g dry weight. The electropherogram detected at 277 nm showed the migration time of leonurine at 6.2 min. The developed CE was found to be valid for leonurine quantification in L. sibiricus ethanolic extract. The contents of leonurine in 15 crude drugs ranged from 0.79 to 4.23 mg/g with the average of 2.38 ± 1.10 mg/g dry weight. This study established the pharmacognostic specification of L. sibiricus crude drug in Thailand with special reference to a bioactive compound, leonurine. CE was beneficial technique for the analysis of leonurine in L. sibiricus aerial parts.

In vitro cytotoxic, genotoxic, and antityrosinase activities of Clitoria macrophylla root.

Clitoria macrophylla Wall. (Leguminosae), locally known as Non-tai-yak or An-chan-pa, commonly distributed in tropical nations and Southeast Asia. Regarding traditional Thai medical system, C. macrophylla roots carry out a potential in dermatology. Its roots are also used as insecticide in agriculture and animal farming. Moreover, clitoriacetal is the major component that can be detected in C. macrophylla root. This research aimed to assess the efficacy of C. macrophylla root extract and clitoriacetal for its anticancer and antityrosinase activities as well as to assess in vitro safety potential for its cytotoxic and genotoxic effects. C. macrophylla root and clitoriacetal were tested by brine shrimp lethality, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, comet assay, and antityrosinase activity. C. macrophylla root, clitoriacetal, and rotenone demonstrated the toxicity against brine shrimp nauplii with LC50 of 332.15, 136.54, and 0.15 µg/mL, respectively. C. macrophylla root and clitoriacetal showed cytotoxic potential against breast ductal carcinoma (BT-474), liver hepatoblastoma (Hep-G2), and colon adenocarcinoma (SW-620). At 100 µg/mL, the percent DNA damage of C. macrophylla root and clitoriacetal was 37.84% and 36.01%, respectively. C. macrophylla root and clitoriacetal were able to inhibit the tyrosinase enzyme with IC50 of 12.27 and 7.30 mg/mL, respectively, which less effective than glutathione (positive control). The present study revealed the in vitro biological activities of C. macrophylla root and its clitoriacetal constituent which proposed the scientific evidences in efficacy and safety evaluation including in vitro cytotoxicity, DNA damage as well as antityrosinase activities.

Quantification of chlorogenic acid, rosmarinic acid, and caffeic acid contents in selected Thai medicinal plants using RP-HPLC-DAD

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 ºC 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.

Quality evaluation with reference to clitoriacetal and in vitro antioxidant activities of Clitoria macrophylla root.

Clitoria macrophylla Wall. (syn. Clitoria hanceana Hemsl.), is commonly known in Thai as Nontai-yak, An-chan-pa, or Ueang-chan-pa, which belongs to Leguminosae family. According to traditional Thai medicine, the root has been used for the treatment of skin diseased as well as for pest control in horticulture and animal husbandry. The aim of this study is to investigate for the pharmacognostic specification, the clitoriacetal content, and in vitro antioxidant capacities of C. macrophylla roots from 12 different sources throughout Thailand. Clitoriacetal content was quantitatively analyzed by thin-layer chromatography (TLC) - densitometry with winCATS software and TLC image analysis with ImageJ software. Antioxidant activities were evaluated by 2, 2-diphenyl-1-picrylhydrazyl radical scavenging, ferric reducing antioxidant power assay, ß-carotene bleaching assay, total phenolic, and total flavonoid contents. The pharmacognostic specification of C. macrophylla roots in Thailand was established. The loss on drying, total ash, acid-insoluble ash, and water contents should be not more than 6.40%, 12.29%, 8.89%, and 8.16% of dry weight, whereas ethanol and water-soluble extractive values should be not less than 4.95% and 14.72% of dry weight, respectively. Furthermore, the clitoriacetal content of C. macrophylla roots determined by TLC-densitometry and TLC image analysis was found to be 2.20 ± 1.31 and 2.22 ± 1.16 g/100 g of dry roots, respectively. The clitoriacetal contents of both methods were not significantly different using paired t-test. Moreover, the ethanolic extract of C. macrophylla roots showed its antioxidant potential compared to the standard butylated hydroxyl toluene and quercetin.

Quality evaluation of Kaempferia parviflora rhizome with reference to 5,7-dimethoxyflavone.

Kaempferia parviflora Wall. ex Baker is a medicinal plant found in the upper Northeastern regions of Thailand, which belongs to Zingiberaceae family. The present study aims to investigate the standardization parameters, to analyze chemical constituents of volatile oil by gas chromatography-mass spectrometry, and to determine the content of 5,7-dimethoxyflavone in K. parviflora rhizomes by thin-layer chromatography (TLC)-densitometry compared to TLC image analysis. K. parviflora rhizomes from 15 different sources throughout Thailand were investigated for morphological and pharmacognostic parameters. 5,7-Dimethoxyflavone contents were determined by TLC-densitometry with winCATS software and TLC image analysis with ImageJ software. The mobile phase for TLC development consisted of toluene: chloroform: Acetone: formic acid (5: 4: 1: 0.2). For the Results, the pharmacognostic parameters of K. parviflora rhizome were demonstrated. The loss on drying, total ash, acid-insoluble ash, water content, volatile oil content, ethanol, and water-soluble extractive values were found to be 8.979 ± 0.041, 5.127 ± 0.060, 2.174 ± 0.092, 9.291 ± 0.458, 0.028 ± 0.003, 5.138 ± 0.092, and 8.254 ± 0.191 g/100 g of dry weight, respectively. K. parviflora volatile oil showed the major components of α-copaene, dauca-5, 8-diene, camphene, ß-pinene, borneol, and linalool. The 5,7-dimethoxyflavone content of K. parviflora rhizomes determined by TLC-densitometry and TLC image analysis were found to be 2.15 ± 0.64 and 1.96 ± 0.51 g/100 g of dry rhizomes, respectively. The 5,7-dimethoxyflavone contents of both methods were not significantly different (P > 0.05) using paired t-test.

Leptocarpus disjunctus prolongs sleeping time and increases nonrapid eye movement sleep with additional anxiolytic capacity.

Leptocarpus disjunctus Mast. (Restionaceae) is an edible plant which has indigenous warnings regarding its side effects which can manifest as dizziness. This study investigated hypnotic and anxiolytic properties using several animal models. Anxiolytic activities were evaluated using locomotor determination by elevated plus-maze test, open-field test, and rotarod performance test. Hypnotic activities were performed using pentobarbital sodium-induced sleeping time test. Sleep architecture and quality were obtained from sleep-wake analysis and nonrapid eye movement (NREM) delta activity using electroencephalography. An ethanolic extract of L. disjunctus indicated effective potencies for hypnotic test, locomotor activities, and sleep-wake analysis. Ethanolic extract showed a dose relationship with sleeping time for pentobarbital-induced sleeping time test (P < 0.01) and also an antagonistic effect on shortening in sleep time induced by flumazenil. The consort significantly decreased locomotor activities among animals undergoing elevated plus-maze test, open-field test, and rotarod performance test, whereas sleep-wake analysis showed that sleeping time and NREM sleep increased. Ethanolic extract of L. disjunctus was shown to be anxiolytic, with the possibly of benzodiazepine-like hypnotic activity.

Pharmacognostic Specification, Chlorogenic Acid Content, and In vitro Antioxidant Activities of Lonicera japonica Flowering Bud.

BACKGROUND: Lonicera japonica Thunb. or Japanese Honeysuckle has been widely used in traditional medicine for antipyretic. OBJECTIVE: To establish the pharmacognostic specification of L. japonica flowering bud in Thailand and to determine its chlorogenic acid content and in vitro antioxidant activities. MATERIALS AND METHODS: Dried L. japonica flowering bud from 15 various herbal drugstores throughout Thailand were investigated for pharmacognostic specification. Their chlorogenic acid contents were quantitatively analyzed by thin layer chromatography (TLC) densitometry with winCATS software. The mobile phase for TLC development consisted of ethyl acetate: formic acid: acetic acid: water (10:1.1:1.1:2.6). Antioxidant activities were investigated by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, ferric ion reducing antioxidant power assay, nitric oxide scavenging assay, and ß-carotene bleaching assays. RESULTS: Qualified L. japonica flowering bud in Thailand was presented that the contents of loss on drying, total ash, acid-insoluble ash, and water should not be >10.11%, 6.59%, 1.14%, and 10.82% by weight, respectively. The ethanol and water soluble extractive values should not be < 16.46% and 28.88% by weight, respectively. Chlorogenic acid content in L. japonica flowering bud was found to be 2.24 ± 0.50 g/100 g of crude drug. L. japonica flowering bud showed DPPH and nitric oxide scavenging activities as well as reducing power property. CONCLUSION: This pharmacognostic specification with special reference to the chlorogenic acid content can be used for quality control of L. japonica flowering bud in Thailand. The potential antioxidant of this crude drug was demonstrated in vitro. SUMMARY: Pharmacognostic specification of Lonicera japonica flowering bud in Thailand has been establishedThe chlorogenic acid content has been quantified by thin layer chromatography-densitometryThe ethanolic extract of L. japonica flowering bud showed antioxidation potential, especially on reducing power property. Abbreviations Used: TLC: Thin layer chromatography, DPPH: 2,2-diphenyl-1-picrylhydrazyl, FRAP: Ferric ion Reducing Antioxidant Power, WHO: World Health Organization, ICH: International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use; LOD: Limit of detection; LOQ: Limit of quantitation; BHT: Butylated hydroxytoluene; FeSO4: Iron(II) sulfate; DMSO: Dimethyl sulfoxide; TPTZ: 2,4,6-tripyridyl-s-triazine.

Pharmacognostic Specifications and Lawsone Content of Lawsonia inermis Leaves.

BACKGROUND: Lawsonia inermis L. has been used as a traditional or folk medicine for the treatment of a wide range of skin infectious diseases. OBJECTIVE: The objective of this study was to determine the pharmacognostic specifications and lawsone contents of L. inermis leaves. MATERIALS AND METHODS: The pharmacognostic specifications of L. inermis leaves from 12 sources were evaluated according to the WHO guideline of quality control method for medicinal plant materials. The lawsone contents were analyzed by thin-layer chromatography (TLC) coupled with densitometry and image analysis. RESULTS: Microscopic evaluation of L. inermis powders showed the fragment of mesophyll, fragment of parenchyma, epidermis layer with stomata, and the rosette crystal of calcium oxalate. Physicochemical parameters revealed that total ash, acid-insoluble ash, loss on drying, and water content should be not <6.98, 1.12, 8.08, and 9.86% of dried weight, respectively, whereas ethanol and water extractive values should be not < 19.67 and 23.06% of dried weight, respectively. The content of lawsone in L. inermis leaves by TLC-densitometry was found to be 0.76 ± 0.05 g/100 g of dried crude drug, whereas the lawsone content evaluation by TLC image analysis was found to be 0.87 ± 0.11 g/100 g of dried crude drug. The validation of the methods revealed that both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation in L. inermis. CONCLUSION: The pharmacognostic specifications could be used as the standardization data of L. inermis leaves, and the development of TLC method could be applied to determine lawsone content in this plant material. SUMMARY: The pharmacognostic specification of Lawsonia inermis leaves could be used as the standardization data of L. inermis leaves in Thailand.Both TLC-densitometry and TLC image analysis showed a good sensitivity and accuracy for lawsone quantitation. Abbreviations Used: LOD: Limit of detection; LOQ: Limit of quantitation; RSD: Relative standard deviation; TLC: Thin layer chromatography; UV: Ultraviolet; Rf value: Relative to front value.

Antidiabetic and anticancer activities of Mangifera indica cv. Okrong leaves.

Diabetes and cancer are a major global public health problem. Plant-derived agents with undesirable side-effects were required. This study aimed to evaluate antidiabetic and anticancer activities of the ethanolic leaf extract of Mangifera indica cv. Okrong and its active phytochemical compound, mangiferin. Antidiabetic activities against yeast α-glucosidase and rat intestinal α-glucosidase were determined using 1 mM of p-nitro phenyl-α-D-glucopyranoside as substrate. Inhibitory activity against porcine pancreatic α-amylase was performed using 1 mM of 2-chloro-4 nitrophenol-α-D-maltotroside-3 as substrate. Nitrophenol product was spectrophotometrically measured at 405 nm. Anticancer activity was evaluated against five human cancer cell lines compared to two human normal cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Mango leaf extract and mangiferin exhibited dose-dependent inhibition against yeast α-glucosidase with the IC50 of 0.0503 and 0.5813 mg/ml, respectively, against rat α-glucosidase with the IC50 of 1.4528 and 0.4333 mg/ml, respectively, compared to acarbose with the IC50 of 11.9285 and 0.4493 mg/ml, respectively. For anticancer activity, mango leaf extract, at ≥200 µg/ml showed cytotoxic potential against all tested cancer cell lines. In conclusion, mango leaf possessed antidiabetic and anticancer potential in vitro.

Characterization of Mangifera indica cultivars in Thailand based on macroscopic, microscopic, and genetic characters.

Thai mango cultivars are classified into six groups plus one miscellaneous group according to germplasm database for mango. Characterization is important for conservation and the development of Thai mango cultivars. This study investigated macroscopic, microscopic leaf characteristics, and genetic relationship among 17 cultivars selected from six groups of mango in Thailand. Selected mango samples were obtained from three different locations in Thailand (n = 57). They were observed for their leaf and fruit macroscopic characteristics. Leaf measurement for the stomatal number, veinlet termination number, and palisade ratio was evaluated under a microscope attached with digital camera. DNA fingerprint was performed using CTAB extraction of DNA and inter-simple sequence repeat (ISSR) amplification. Forty-five primers were screened; then, seven primers that amplified the reproducible band patterns were selected to amplified and generate dendrogram by Unweighted Pair-Group Method with Arithmetic Average. These selected 17 Thai mango cultivars had individually macroscopic characteristics based on fruits and leaves. For microscopic characteristics, the stomatal number, veinlet termination number, and palisade ratio were slightly differentiable. For genetic identification, 78 bands of 190-2660 bps were amplified, of which 82.05% were polymorphic. The genetic relationship among these cultivars was demonstrated and categorized into two main clusters. It was shown that ISSR markers could be useful for Thai mango cultivar identification.

Chlorogenic acid content, essential oil compositions, and in vitro antioxidant activities of Chromolaena odorata leaves.

Chromolaena odorata (L.) R. M. King and H. Rob. is a Thai medicinal plant used for the treatment of wounds, rashes, diabetes, and insect repellent. The leaves of C. odorata were collected from 10 different sources throughout Thailand. The chemical constituents of essential oils were hydro-distilled from the leaves and were analyzed by gas chromatography-mass spectrometry. Chlorogenic acid contents were determined by thin-layer chromatography (TLC) - densitometry with winCATS software and TLC image analysis with ImageJ software. The TLC plate was developed in the mobile phase that consisted of ethyl acetate:water:formic acid (17:3:2). Antioxidant activities were examined by 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging and ß-carotene bleaching assays. C. odorata essential oil has shown the major components of pregeijerene, dauca-5, 8-diene, (E)-caryophyllene, ß-pinene, and α-pinene. The chlorogenic acid content of C. odorata leaves was determined by TLC-densitometry and TLC image analysis. Results have shown that TLC-densitometry and TLC image analysis method were not statistically significantly different. DPPH radical scavenging and ß-carotene bleaching assays of ethanolic extract of C. odorata leaves showed its antioxidant potential.

Pharmacognostic evaluation with reference to catechin content and antioxidant activities of pale catechu in Thailand.

Pale catechu, a well-known crude drug, has been widely used for anti-diarrhea. Due to its medicinal usage, this study was performed to evaluate the pharmacognostic and antioxidant properties as well as catechins contents of pale catechu in Thailand. Twenty samples of pale catechu collected from traditional drug stores throughout Thailand were investigated. Antioxidant activities, total phenolic, nontannin phenolic, and total tannin contents were evaluated. (+)-catechin and (-)-epicatechin were quantitatively analyzed by high performance liquid chromatography. The results revealed that most of pale catechu samples were adulterated according to high ash values. Qualified pale catechu in Thailand were demonstrated for their average contents of total ash, acid insoluble ash, loss on drying, and moisture as 5.20 ± 0.19, 1.61 ± 0.17, 13.14 ± 0.10, and 13.20 ± 1.07 g/100 g of dry weight, respectively. The ethanol and water soluble extractive matters were 91.66 ± 5.16 and 44.59 ± 3.18 g/100 g of dry weight respectively. (+)-catechin in theses samples was 478.87 ± 2.77 µg/mg of crude drug, whereas (-)-epicatechin was found to be trace (

In vitro anti-inflammatory, mutagenic and antimutagenic activities of ethanolic extract of Clerodendrum paniculatum root.

Clerodendrum paniculatum L. (Family Verbenaceae) has been used as an antipyretic and anti-inflammatory drug in traditional Thai medicine. This present study investigated the in vitro anti-inflammatory, mutagenic and antimutagenic activities of the ethanolic extract of C. paniculatum (CPE) dried root collected from Sa Kaeo Province of Thailand. Murine macrophage J774A.1 cells were stimulated by lipopolysaccharide (LPS) to evaluate nitric oxide (NO), tumor necrosis factor-α (TNF-α) and prostaglandin E2 (PGE2) production in the anti-inflammatory test while the mutagenic and antimutagenic potential was performed by the Ames test. The outcome of this study displayed that the CPE root significantly inhibited LPS-induced NO, TNF-α, and PGE2 production in macrophage cell line. In addition, the CPE root was not mutagenic toward Salmonella typhimurium strain TA98 and TA100 with and without nitrite treatment. Moreover, it inhibited the mutagenicity of nitrite treated 1-aminopyrene on both strains. The findings suggested the anti-inflammatory and antimutagenic potentials of CPE root.

Pharmacognostic specifications and quantification of (+)-catechin and (-)-epicatechin in Pentace burmanica stem bark.

BACKGROUND: According to Thai traditional medicine, Pentace burmanica Kurz. stem bark has been used as crude drug for treating diarrhea. However, the crude drug is also found susceptible to adulteration. OBJECTIVES: To develop specific standardization parameters of P. burmanica stem bark in Thailand and to determine the (+)-catechin and (-)-epicatechin contents of P. burmanica stem bark by HPLC analysis. MATERIALS AND METHODS: P. burmanica stem barks from various sources throughout Thailand were investigated according to WHO guideline of the pharmacognostic specification. High performance liquid chromatography (HPLC) was performed for (+)-catechin and (-)-epicatechin quantification. RESULTS: Macroscopic evaluation was demonstrated as whole plant drawing. Microscopic evaluation of stem bark powdered drug showed fragment of fibers, resin masses, tannin masses, starch grain, calcium oxalate, and fragment of parenchyma. Physico-chemical parameters revealed that total ash, acid insoluble ash, loss on drying, and water content should be not more than 3.58, 0.50, 8.40, and 9.70% of dry weight respectively; while ethanol and water soluble extractive values should not be less than 21.90 and 19.06% of dry weight respectively. Both (+)-catechin and (-)-epicatechin were existed in P. burmanica ethanolic extract. Owing to the small amount of (+)-catechin, quantitation of its content was omitted. However, (-)-epicatechin contents was found as 59.74 ± 1.69µg/mg of crude extract. CONCLUSION: The pharmacognostic investigations can be used to set the standard parameters of P. burmanica stem bark in Thailand. HPLC method can be applied to determine (+)-catechin and (-)-epicatechin content in plant materials.

Quantitative analysis of oxyresveratrol content in Artocarpus lakoocha and 'Puag-Haad'.

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 F(254) TLC plate (20 x 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.

Molecular analysis of the genus Mitragyna existing in Thailand based on rDNA ITS sequences and its application to identify a narcotic species: Mitragyna speciosa.

In Thailand, there are four Mitragyna species; M. speciosa, M. hirsuta, M. diversifolia, and M. rotundifolia. One, M. speciosa, is a narcotic plant and has medicinal importance for its opium-like effect. Since the use of M. speciosa has been forbidden in Thailand, the leaves of M. diversifolia or others are frequently used as substitutes but are not considered as effective. Therefore, accurate authentication of M. speciosa is essential for both medicinal and forensic purposes. The nucleotide sequences of internal transcribed spacers (ITS) and the 5.8S coding region of nuclear ribosomal DNA (rDNA) of the Mitragyna species were analyzed. The whole length of ITS1-5.8S-ITS2 region was 608 bp in M. speciosa, 607 bp in the other species. Nineteen sites of nucleotide substitutions and 3 sites of 1-bp indels were observed, and M. speciosa showed specific sequence differed from the others. Based on the ITS sequences, a distinctive site recognized by a restriction enzyme XmaI in M. speciosa was found and then PCR-restriction fragment length polymorphism (RFLP) analysis was established to differentiate M. speciosa from the others. By the method, a 409-bp PCR fragment of ITS1-5.8S (partial) rDNA region from M. speciosa was cleaved into two fragments of 119 bp and 290 bp while the other species remained undigested. This method provides an effective and accurate identification of M. speciosa.

Chiral separation in capillary electrophoresis using dual neutral cyclodextrins: theoretical models of electrophoretic mobility difference and separation selectivity.

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.