National Dope Testing Laboratory

Delhi, India

National Dope Testing Laboratory

Delhi, India
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Priya N.G.,University of Delhi | Ojha A.,International Center for Genetic Engineering and Biotechnology | Kajla M.K.,University of Wisconsin - Madison | Raj A.,National Dope Testing Laboratory | Rajagopal R.,University of Delhi
PLoS ONE | Year: 2012

Helicoverpa are important polyphagous agricultural insect pests and they have a worldwide distribution. In this study, we report the bacterial community structure in the midgut of fifth instar larvae of Helicoverpa armigera, a species prevalent in the India, China, South Asia, South East Asia, Southern & Eastern Africa and Australia. Using culturable techniques, we isolated and identified members of Bacillus firmus, Bacillus niabense, Paenibacillus jamilae, Cellulomonas variformis, Acinetobacter schindleri, Micrococcus yunnanesis, Enterobacter sp., and Enterococcus cassiliflavus in insect samples collected from host plants grown in different parts of India. Besides these the presence of Sphingomonas, Ralstonia, Delftia, Paracoccus and Bacteriodetes was determined by culture independent molecular analysis. We found that Enterobacter and Enterococcus were universally present in all our Helicoverpa samples collected from different crops and in different parts of India. The bacterial diversity varied greatly among insects that were from different host plants than those from the same host plant of different locations. This result suggested that the type of host plant greatly influences the midgut bacterial diversity of H. armigera, more than the location of the host plant. On further analyzing the leaf from which the larva was collected, it was found that the H. armigera midgut bacterial community was similar to that of the leaf phyllosphere. This finding indicates that the bacterial flora of the larval midgut is influenced by the leaf surface bacterial community of the crop on which it feeds. Additionally, we found that laboratory made media or the artificial diet is a poor bacterial source for these insects compared to a natural diet of crop plant. © 2012 Gayatri Priya et al.


Rahman N.,Aligarh Muslim University | Anwar N.,Anand Engineering College | Kashif M.,National Dope Testing Laboratory | Hoda M.N.,Georgia Regents University | Rahman H.,Aligarh Muslim University
Journal of the Mexican Chemical Society | Year: 2011

A simple and sensitive kinetic spectrophotometric method was developed for the determination of labetalol (LBT) hydrochloride. The method was based on the kinetic investigation of the oxidation of the drug with alkaline potassium permanganate at room temperature (25 ± 1°C). The increase in absorbance of coloured manganate ions was measured at 605 nm. All experimental variables affecting the development of the colour were investigated and optimized. The initial rate and fixed time (at 6 minute) methods were adopted for determining the drug concentration. The calibration graphs were linear in the concentration ranges of 2-14 μg mL-1 and 1-10 μg mL-1, using the initial rate and fixed time methods, respectively. The method was successfully applied to the determination of labetalol in laboratory made tablets and commercial tablets. The results were validated statistically and through recovery studies. © 2011, Sociedad Química de México.


Ahi S.,National Dope Testing Laboratory | Beotra A.,National Dope Testing Laboratory | Dubey S.,National Dope Testing Laboratory | Upadhyay A.,National Dope Testing Laboratory | Jain S.,National Dope Testing Laboratory
Drug Testing and Analysis | Year: 2012

The use of prednisolone and prednisone is prohibited by the World Anti-Doping Agency (WADA) due to their performance-enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11-oxo metabolite) [M-1], 6-β-OH-prednisolone [M-2], 20-β-OH-prednisolone [M-3], 20-α-OH-prednisolone [M-4], 20-α-OH-prednisone [M-5], 20-β-OH-prednisone [M-6], 2 tetrahydro epimers of 20-β-OH-prednisolone [M-7], 2 tetrahydro epimers of 20-α-OH-prednisolone [M-8], 2 tetrahydro epimers of 20-β-OH-prednisone [M-9], and 2 tetrahydro epimers of 20-α-OH-prednisone [M-10]. Prednisolone was administered in 10-, 20-, and 40-mg dosage to healthy volunteers to study detection of various metabolites. The parent, M-1, M-2, and M-3 could be detected up to 72h while rest of the metabolites were detectable up to 24h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes. © 2012 John Wiley & Sons, Ltd.


PubMed | National Dope Testing Laboratory
Type: Journal Article | Journal: Drug testing and analysis | Year: 2012

The use of prednisolone and prednisone is prohibited by the World Anti-Doping Agency (WADA) due to their performance-enhancing effect. The purpose of the present work was to explore the possibility of identification and detection of various metabolites of prednisolone by liquid chromatography-tandem mass spectrometry (LC-MS/MS) in excretion study samples. Ten metabolites of prednisolone could be identified namely prednisone (11-oxo metabolite) [M-1], 6--OH-prednisolone [M-2], 20--OH-prednisolone [M-3], 20--OH-prednisolone [M-4], 20--OH-prednisone [M-5], 20--OH-prednisone [M-6], 2 tetrahydro epimers of 20--OH-prednisolone [M-7], 2 tetrahydro epimers of 20--OH-prednisolone [M-8], 2 tetrahydro epimers of 20--OH-prednisone [M-9], and 2 tetrahydro epimers of 20--OH-prednisone [M-10]. Prednisolone was administered in 10-, 20-, and 40-mg dosage to healthy volunteers to study detection of various metabolites. The parent, M-1, M-2, and M-3 could be detected up to 72h while rest of the metabolites were detectable up to 24h after drug administration. The detection of newer metabolites of the drug can further be used for confirmatory purposes.


PubMed | National Dope Testing Laboratory
Type: Journal Article | Journal: Drug testing and analysis | Year: 2010

The abuse of stanozolol is quite widespread in Indian sport. Its analysis is challenging and this has led to the development of new methods to improve its detection. A method was developed and validated for the detection of the three main monohydroxylated metabolites of stanozolol. The excretion profile of these metabolites was studied in four healthy male volunteers. The excretion study samples, after a single oral dose of drug, showed that 3-OH-stanozolol was excreted at the highest concentration, followed by 16beta-OH stanozolol, with 4beta-OH stanozolol as the least excreted. Ninety-eight old doping samples with adverse analytical findings for 3-OH-stanozolol were reanalysed using this method. This showed 3-OH-stanozolol and 16beta-OH stanozolol in all the 98 samples whereas 4beta-OH-stanozolol was identified in 90 samples. The percentage of positive identifications of stanozolol in Indian sportspeople has increased markedly in the last five years, from 31.9% in 2004 to 81.8% in 2009; however, this may be due to the more effective detection of stanozolol metabolites. It can thus be concluded that the marked increase in percent positive of stanozolol in Indian sportspersons in 2009 may be due to the improved detection by a more effective LCMS/MS method.


PubMed | National Dope Testing Laboratory
Type: Journal Article | Journal: Indian journal of pharmacology | Year: 2010

The use of performance enhancing substances is banned in sports by the World Anti-Doping Agency (WADA). Though most prohibited substances can be detected by GC/MS, inclusion of corticosteroids and designer drugs has made it essential to detect these critical doping agents on LC/MS/MS due to their better separation and detection.A common extraction procedure for the isolation of acidic, basic and neutral drugs from urine samples was developed. A total of 28 doping drugs were analyzed on API 3200 Triple quadrupole mass spectrometer using C18 column in atmospheric pressure electrospray ionization. The mobile phase composition was a mixture of 1% formic acid and acetonitrile with gradient time period.The method developed was very sensitive for detection of 28 doping agents. The linearity was performed for each drug and the total recovery percentage ranged from 57 to 114. Limit of detection is found to be 0.5 ng/ml for carboxy finasteride and 1-5 ng/ml for other drugs. The method was successfully used to detect positive urine samples of 3-OH-stanozolol, methyl phenidate, mesocarb, clomiphene metabolite and carboxy finasteride.The method developed based on controlled pH extraction method and HPLC-mass spectrometry analysis allowed better identification and confirmation of glucocorticosteroids and a few other drugs in different categories. The validated method has been used successfully for testing of 1000 In-competition samples. The method helped in detection of chemically and pharmacologically different banned drugs in urine in a single short run at a minimum required performance limit set by WADA.


PubMed | National Dope Testing Laboratory
Type: Journal Article | Journal: Indian journal of pharmacology | Year: 2010

Adrafinil and modafinil have received wide publicity and have become controversial in the sporting world when several athletes were discovered allegedly using these drugs as doping agents. By acknowledging the facts, the World Anti-Doping Agency (WADA) banned these drugs in sports since 2004. The present study explores the possibility of differentiating adrafinil and modafinil and their major metabolites under electron impact ionization in gas chromatograph-mass spectrometer (GC-MSD) and electrospray ionization in liquid chromatograph-mass spectrometer (LC-MS/MS) by studying the fragmentation pattern of these drugs.Adrafinil, modafinil and their major metabolite, modafinilic acid were analyzed on EI-GC-MSD and ESI-LC-MS/MS using various individual parameters on both the instruments. The analytical technique and equipment used in the analysis were an Agilent 6890N GC with 5973 mass selective detector for the GC-MSD analysis and an Agilent 1100 HPLC with API-3200 Triple quadrupole mass spectrometer for the LC-MS/MS analysis. Validation of both methods was performed using six replicates at different concentrations.The results show that adrafinil, modafinil and their major metabolite modafinilic acid could be detected as a single artifact without differentiation under EI-GC-MSD analysis. However, all drugs could be detected and differentiated under ESI-LCMS/MS analysis without any artifaction. The GC-MSD analysis gives a single artifact for both the drugs without differentiation and thus can be used as a marker for screening purposes. Further, the Multiple Reaction Monitoring (MRM) method developed under LC-MS/MS is fit for the purpose for confirmation of suspicious samples in routine sports testing and in forensic and clinical analysis.

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