Subhash C. Singh

A promising anticancer and antimalarial component from the leaves of Bidens pilosa.

Bidens pilosa is used in folk medicine for various applications due to the presence of polyacetylenes, flavonoids, terpenoids, phenylpropanoids and others. Bioactivity-guided fractionation of different extracts of B. pilosa leaf showed potential in vitro anticancer and antimalarial activity and led to the identification of a potential marker compound, phenyl-1,3,5-heptatriyne. Erythrocyte osmotic fragility experiments revealed the various extracts as well as the marker components toxicity profiles on normal blood cells.

HPLC DETERMINATION OF VASICINE AND VASICINONE IN ADHATODA VASICA WITH PHOTO DIODE ARRAY DETECTION

A high performance liquid chromatographic method for the determination of the quinazoline alkaloids vasicine (1) and vasicinone (2) in Adhatoda vasica plant extract is reported. Peak purity and similarity of 1 and 2 have been studied using photodiode array detector (PDA). Effects of different solvents have been studied for the extraction of 1 and 2 in A. vasica plant, and methanol was found to give the maximum extraction of compounds 1 and 2. The method is simple, sensitive, rapid, and reproducible for the quantitation of pharmacologically important alkaloids vasicine and vasicinone. The separation of 1 and 2 was performed with acetonitrile–phosphate buffer (pH maintained to 3.9 using glacial acetic acid) (15:85) using a Hibar Merck make C18 column.

RP-LC determination of oleane derivatives in Terminalia arjuna

A rapid sensitive and reproductive reversed phase high performance liquid chromatographic method with photo diode arrray detection is described for the simultaneous quantification of major oleane derivatives: arjunic acid (4), arjunolic acid (3), arjungenin (2) and arjunetin (1) in Terminalia arjuna extract. The method involves the use of a Waters Spherisorb S10 ODS2 column (250 x 4.6 mm, I.D., 10 microm) and binary gradient mobile phase profile. The various other aspects of analysis viz. Extraction efficiency, peak purity and similarity were validated using a photo diode array detector.

4α-Methyl-24β-ethyl-5α-cholesta-14,25-dien-3β-ol and 24β-ethylcholesta-5, 9(11), 22E-trien-3β-ol, sterols from Clerodendrum inerme

From the aerial parts of Clerodendrum inerme, two new sterols (4alpha-methyl-24beta-ethyl-5alpha-cholesta-14, 25-dien-3beta-ol and 24beta-ethylcholesta-5, 9(11), 22E-trien-3beta-ol) and a new aliphatic ketone (11-pentacosanone) were isolated together with another known aliphatic ketone (6-nonacosanone) and a diterpene (clerodermic acid). The structure elucidations were based on analyses of physical and spectroscopic data.

Composition of linalool rich essential oil from Lippia alba grown in Indian plains

A field crop of Lippia alba (Mill.) N.E. Br. LAC-2 genotype was raised through stem cuttings at Lucknow. The essential oil yields obtained upon hydrodistillation of leaves harvested between May 1998 and May 1999 varied from 0.6% to 0.8% on fresh weight basis. The GC and GC/MS analyses of the essential oils led to the identification of 15 compounds totalling 85% of the oil: the identified compounds included 4 monoterpene hydrocarbons (2%), 8 oxygenated monoterpenes (82%) and 3 sesquiterpene hydrocarbons (1%). The major constituent of the oil was linalool (65%). Copyright

Antifilarial diarylheptanoids from Alnus nepalensis leaves growing in high altitude areas of Uttarakhand, India

Lymphatic filariasis continues to be a major health problem in tropical and subtropical countries. A macrofilaricidal agent capable of eliminating adult filarial parasites is urgently needed. Platyphyllenone (A), alusenone (B), hirustenone (C) and hirsutanonol (D) are important biologically active diarylheptanoids present in Alnus nepalensis. In the present study, we report the antifilarial activity in diarylheptanoids isolated from the leaves of A. nepalensis. Out of four compounds (A-D) tested in vitro one has shown promising anti-filarial activity both in vitro and in vivo studies. This is the first ever report on antifilarial efficacy of a compound of the plant and warrants further studies around this scaffold. In addition, a sensitive, selective and robust densitometric high-performance thin-layer chromatographic method was developed and validated for the above four biomarker compounds. The separation was performed on silica gel 60F(254) high-performance thin layer chromatography plates using chloroform:methanol (9:1, v/v) as mobile phase. The quantitation of marker compounds was carried out using densitometric reflection/absorption mode at 600 nm after post-chromatographic derivatization using vanillin-sulfuric acid reagent. The method was validated for peak purity, precision, robustness, limit of detection (LOD) and quantitation (LOQ) etc., as per the International Conference on Harmonization (ICH) guidelines.

Wide spectrum antibacterial and antifungal activities in the seeds of some coprophilous plants of north Indian plains

In a survey at Lucknow, India, the seedlings of plant species which are prescribed in the Indian traditional system of medicine for a variety of infectious diseases were found to predominate on fresh or decomposing cattle dung, a harsh medium for plant growth due to high microbial load and other abiotic factors. Plants of most of the common species did not occur on the cattle dung heaps. It was hypothesized that plant species which are able to grow on cattle dung may have antimicrobial compounds in their seeds to protect them from microbial attack. In confirmation, the seeds of 15 of the coprophilous (kopros--dung, philein--to love) plant species, identified as occurring most frequently on fresh/decomposing cattle dung were directly tested against eight bacterial and three fungal strains. Interestingly, seeds of all the examined species exhibited antimicrobial activity. The seeds of the species found more frequently on the cattle dung heaps possessed higher levels of antimicrobial activities.

Genetic and Chemical Diversity of High Mucilaginous Plants of Sida Complex by ISSR Markers and Chemical Fingerprinting

A method was developed based on multiple approaches wherein DNA and chemical analysis was carried out toward differentiation of important species of Sida complex that is being used for commercial preparation. Isolated DNA samples were successfully performed through PCR amplification using ISSR markers and degree of genetic diversity among the different species of Sida is compared with that of chemical diversity. For genetic fingerprint investigation, selected 10 ISSR primers generating reproducible banding patterns were used. Among the total of 63 amplicons, 62 were recorded as polymorphic, genetic similarity index deduced from ISSR profiles ranged from 12 to 51%. Based on similarity index, S. acuta and S. rhombifolia found to be most similar (51%). High number of species-specific bands played pivotal role to delineate species at genetic level. Investigation based on HPTLC fingerprints analysis revealed 23 bands representing to characteristic chemicals and similarity index ranged from 73 to 91%. Prominent distinguishable bands were observed only in S. acuta, while S. cordifolia and S. rhombifolia shared most bands making them difficult to identify on chemical fingerprint basis. This report summarizes the genotypic and chemotypic diversity and the use of profiles for authentication of species of Sida complex.

Simple and reliable methods for the determination of three steroidal glycosides in the eight species of Solanum by reversed-phase HPLC coupled with diode array detection.

INTRODUCTION Solanum species are important ingredients of many traditional Indian medicines and thus the quality control of their herbal formulations is of paramount concern. OBJECTIVE To establish a simple and effective high-performance liquid chromatographic (HPLC) method to evaluate the quality of Solanum species and their herbal formulations. METHODOLOGY A rapid, simple, sensitive, robust and reproducible HPLC method was developed for the determination of three steroidal glycosides (SG); indioside D, solamargine and α-solanine in eight species of the genus Solanum. The analytes were separated on a monolithic performance RP-18e column (100 mm × 4.6 mm i.d.) using a gradient elution of acetonitile-water containing 0.1% trifluoroacetic acid (TFA) as the mobile phase with a flow rate 0.4 mL/min and UV detection at λ 210 nm. RESULTS The method was linear over the range 3-15 µg/mL (r > 9994). Accuracy, precision and repeatability were all within the required limits. The mean recoveries measured at the three concentrations were higher than 98.8% with RSD < 2% for the targets. CONCLUSION The established method is simple and can be used as a tool for quality control of plant material or herbal formulation containing SG.

A transcriptomic approach for exploring the molecular basis for dosha-balancing property-based classification of plants in Ayurveda

In Ayurveda, a healthy body is defined by a balance among the three doshas (Vata, Pitta, Kapha) and ailments result due to imbalances among them. It prescribes specific plant parts/tissues collected in a season-specific manner for curing dosha-related imbalances but the plants prescribed for treating a particular dosha imbalance belong to taxonomically diverse families and often contain similar classes of phytomolecules, making it difficult to provide a phytochemical validation for any similarity that might exist among them. This exploratory study hypothesised that plants of the same dosha-curing group may have similarity at the transcript level. For proving/disproving the hypothesis, cDNA–AFLP and specific expression subset analysis (SESA) were carried out on the Ayurveda-defined active tissues of four representative plants each of the three dosha-balancing groups. cDNA–AFLP analyses indicated that even though the plants belonging to a particular dosha-group may widely differ at the transcript level, there is a small fraction of transcripts that is monomorphic among their active tissues. SESA (Tester—active tissue cDNA; Driver—cDNA from other major tissue[s]) generated 803 subtractive ESTs from the twelve plants that yielded 150 unigenes upon assembly (of ESTs from each plant separately). Cross-plant EST assembly for plants in the same dosha group also corroborated the results. Although a distinct pattern of transcripts was not observed across all the plants in a particular dosha group, some commonalities were obtained that need further characterization towards searching for the hitherto elusive similarity among plants of the same group.