R.C. Murthy

Male reproductive toxicity of sodium arsenite in mice

The effect of chronic oral exposure to arsenic on male mouse testicular and accessory sex organ weights, sperm parameters and testicular marker enzymes was studied. In addition, the distribution of arsenic in reproductive organs was measured using atomic absorption spectrophotometry. Sodium arsenite administered to mice (Mus musculus) via drinking water at a dose of 53.39 βmol/L (4 ppm As) for 365 days caused a decrease in the absolute and relative testicular weight. However, epididymal and accessory sex organ weight was similar to control. The activities of marker testicular enzymes such as sorbitol dehydrogenase, acid phosphatase and 17β-hydroxysteroid dehydrogenase (17β-HSD) were significantly decreased, but those of lactate dehydrogenase and γ-glutamyl transpeptidase (γ-GT) were significantly increased. A decrease in sperm count and sperm motility, along with an increase in abnormal sperm, was observed in arsenite-exposed mice. A significant accumulation of arsenic in testes, epididymis, seminal vesicle and prostate gland was observed in treated animals. Thus long term exposure (365 days) at the dose level of 53.39 μmol/L sodium arsenite (4 ppm As), to which human beings are likely to be exposed via drinking water, may cause testicular and spermatotoxic effect.

Embryo and fetotoxicity of hexavalent chromium: a long-term study

Ingestion of chromium(VI) (250, 500 or 750 ppm as potassium dichromate, K2Cr2O7) through drinking water by female rats for 3 months prior to gestation was toxic to embryo and fetus. There was a significant reduction in number of implantations and number of fetuses and an increase in number of resorptions and pre-implantation and post-implantation losses. No significant visceral abnormality was found. The increase in the number of subdermal hemorrhagic patches on the thorax and abdomen was significant. Skeletal abnormality in the form of reduced ossification in parietal, interparietal and caudal bones was observed in fetuses. Chromium levels in the blood of mothers, placenta and fetuses showed a significant increase. Duration of the estrous cycle was also increased significantly. The study revealed that long-term chromium exposure in rats did not cause embryo and fetotoxicity in a duration-dependent manner compared to short-term treatment as observed earlier. A possible explanation could be that, in the 90-day study, the female rats did not mate for three estrous cycles, thus giving time for clearance of a sizable amount of chromium from their bodies.

Neuromelanin in manganese-exposed primates

Monkeys developed muscular weakness and rigidity of the lower limbs after 18 months exposure to manganese. These are neurological signs typical of chronic manganese intoxication. Marked neuronal degeneration with depigmentation was noticed in the region of substantia nigra. Significance of depigmentation in relation to the depletion in the contents of brain dopamine in chronic manganese intoxication has been discussed.

Chromium induced teratogenicity in female rat

Exposure to chromium (VI) (250, 500 and 750 ppm as potassium dichromate) via drinking water pregestationally in rats revealed embryo- and fetotoxic effects in the form of a significant reduction in the number of implantations and number of fetuses. An increase in the number of resorptions, pre-implantation and post-implantation loss in chromium (VI)-treated mothers was also observed. No significant visceral abnormality was found. A significant increase in sub-dermal hemorrhagic patches on thoracic and abdominal areas was found. Skeletal abnormality in the form of reduced ossification in parietal, interparietal and caudal bones was found in the fetuses of chromium (VI)-treated mothers. Chromium levels in blood, placenta and fetuses were found to be significantly increased in the 500 ppm and 750 ppm dosed groups. The duration of estrus cycle was significantly altered after chromium (VI) exposure. This study suggests that chromium exposure in rat causes a lower degree of toxicity than in mice as observed in our earlier studies.

Embryotoxicity of orally administered chromium in mice: Exposure during the period of organogenesis

Administration of chromium (VI)(250, 500 and 750 ppm as potassium dichromate) via drinking water during organogenesis (days 6-14 of gestation) in mice revealed embryo- and fetotoxic effects. Reduced fetal weight, retarded fetal development, number of fetuses (live and dead) per mother and high incidences of dead fetuses and resorptions in treated mothers in the highest dosed group were evident. No significant gross structural abnormalities were observed in any of the fetuses of chromium (VI)-treated mothers. Significant incidences of reduced ossification were found in the highest dosed group. Chromium levels were increased in a dose-dependent manner in maternal blood, placenta and fetuses. The present study suggests a risk to the developing embryo if the mother is exposed to a sufficiently high concentration of chromium (VI) through drinking water during the period of organogenesis.

Behavioral and neurochemical changes in rats simultaneously exposed to manganese and lead

Groups of rats were exposed simultaneously to manganese chloride (3 mg Mn2+/ml water) through drinking water and lead acetate intraperitoneally at dosages of 5.0, 8.0 and 12.0 mg Pb2+/kg daily for a period of 14 days. The magnitude of changes in the behavioral pattern, contents of biogenic amines and accumulation of lead in the brain of rats simultaneously exposed to the two metals was significantly greater than observed in rats after exposure to either of the metals alone. A definite dose-response relationship was, however, noticed only with the changes in the motoractivity, norepinephrine, 5-hydroxytryptamine levels and in the accumulation of lead in rats simultaneously exposed to manganese and lead. The lowering in the contents of norepinephrine after combined treatment was found to be related with the decrease in the motoractivity in the rats. The exact role of depression in the levels of dopamine and 5-hydroxytryptamine in inducing marked impairment in learning ability and increased aggressive behavior in rats after the combined exposure to manganese and lead could not be ascertained. The overall analysis of the data indicated that the simultaneous exposure to manganese and lead, particularly with highest dose of the latter, may produce serious derangements in the behavioral pattern and levels of biogenic amines in the brain of rats.

Embryotoxicity and fetotoxicity of orally administered hexavalent chromium in mice

The embryotoxic and fetotoxic potential of hexavalent chromium (Cr+6) in mice was investigated by administering 250, 500, and 1000 ppm of potassium dichromate daily through drinking water during the entire gestation period. An increase in embryonic deaths was observed; however, in the mothers treated with the highest dose, there was complete absence of implantation sites. No major abnormality was observed in the fetuses except that Cr+6 exposure increased the incidences and types of external and skeletal malformations. It is concluded that oral exposure to Cr+6 causes dose-dependent embryolethal effects in mice.

Ovarian dysfunction in mice following chromium (VI) exposure

Chromium (VI) was given through drinking water in two sets of adult Swiss albino female mice in three doses; 250 ppm, 500 ppm and 750 ppm for 20 days in set 1 and 0.05 ppm, 0.5 ppm and 5.0 ppm in set II for 90 days. At the termination of the treatment, the animals of both the sets were euthanized for histopathology, follicle counting, counting of the superovulated ova, duration of estrus cycle and for ultrastructural studies. Ovaries of the highest dose group (750 ppm) showed large numbers of atretic follicles and congestion in stromal tissue compared to the rest of the treated groups. Also, there was a dose-dependent reduction in the number of follicles at different stages of their maturation. The number of ova recovered from superovulated chromium (VI)-treated animals showed significant decreases in the 500 and 750 ppm dosed groups compared to lower dosed (250 ppm) and control groups. The duration of estrus cycle increased in highest dosed (750 ppm) group. A dose-dependent increase in blood chromium level was also seen in treated mice. Ultrastructural observations revealed disintegrated cell membranes of two layered follicular cells and altered villiform mitochondria in thecal cells of 5 ppm dosed group. From the study it was concluded that ovarian physiology and rate of ovulation might be altered if females are exposed to sufficiently high chromium through oral route.

Simultaneous derivatisation and preconcentration of parabens in food and other matrices by isobutyl chloroformate and dispersive liquid-liquid microextraction followed by gas chromatographic analysis.

A simple, rapid and economical method has been proposed for the quantitative determination of parabens (methyl, ethyl, propyl and butyl paraben) in different samples (food, cosmetics and water) based on isobutyl chloroformate (IBCF) derivatisation and preconcentration using dispersive liquid-liquid microextraction in single step. Under optimum conditions, solid samples were extracted with ethanol (disperser solvent) and 200 μL of this extract along with 50 μL of chloroform (extraction solvent) and 10 μL of IBCF was rapidly injected into 2 mL of ultra-pure water containing 150 μL of pyridine to induce formation of a cloudy state. After centrifugation, 1 μL of the sedimented phase was analysed using gas chromatograph-flame ionisation detector (GC-FID) and the peaks were confirmed using gas chromatograph-positive chemical ionisation-mass spectrometer (GC-PCI-MS). Method was found to be linear over the range of 0.1-10 μg mL(-1) with square of correlation coefficient (R(2)) in the range of 0.9913-0.9992. Limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.029-0.102 μg mL(-1) and 0.095-0.336 μg mL(-1) with a signal to noise ratio of 3:1 and 10:1, respectively.

Development, validation and comparison of two microextraction techniques for the rapid and sensitive determination of pregabalin in urine and pharmaceutical formulations after ethyl chloroformate derivatization followed by gas chromatography–mass spectrometric analysis

The present article reports first time the use of solid-phase microextraction (SPME) and dispersive liquid-liquid microextraction (DLLME) to extract pregabalin (PRG) from urine and pharmaceutical formulations followed by GC-MS analysis after ethyl chloroformate (ECF) derivatization. PRG is an antiepileptic and analgesic drug, which is a structural analogue of γ-amino-butyric acid (GABA). It is approved by Food and Drug Administration (FDA) for the treatment of central nervous system (CNS) disorders and neuropathic pain. Initially PRG was derivatized with ECF in the presence of pyridine at room temperature for 30s. Experimental parameters were investigated for derivatization, SPME and DLLME conditions. The limit of detection (LOD) and limit of quantitation (LOQ) were found to be 0.019 μg/ml and 0.063 μg/ml for SPME and 0.022 μg/ml and 0.075 μg/ml for DLLME respectively. The percentage recovery, in case of SPME was in the range of 83-98% while for DLLME it is in the range of 84-98%. The intra and inter-day precisions were found to be less than 6%. The developed methods after ECF derivatization were found to be simple, fast, efficient and inexpensive. DLLME has several advantages like lesser extraction time and cost effectiveness as compared to SPME. The developed methods may find wide application for the routine determination of PRG in biological as well as in quality control samples of pharmaceutical formulations.