Kidney Of Wistar Rats Research Articles

Effect of A. precatorius Aqueous Seed Extract on the Histology of Kidney, Lungs and Intestines of Wistar Rats

Effect of A. precatorius Aqueous Seed Extract on the Histology of Kidney, Lungs and Intestines of Wistar Rats

Ameliorative effects of Embilica officinalis and Rosmarinus officinalis on cadmium-induced oxidative stress in Wistar rats

The study was designed to assess the antioxidative potential of three dietary antioxidants, vitamin C, Rosemary (Rosmarinus officinalis) leaf extract (10% carnosic acid) and Amla ( Embilica officinalis ) fruit extract (aqueous) on cadmium-induced oxidative stress and the related histopathology of the liver and kidney of Wistar rats. Cadmium was administered as cadmium chloride (15 mg/kg body weight/day) for 4 weeks with five groups co-treated with the antioxidants, vitamin C (100 mg/kg/day) Rosemary leaf extract (15 and 30 mg/kg/day) and amla fruit extract (100 and 200 mg/kg/day). The results showed that cadmium induced a significant increase in both kidney and liver malondialdeyhde (MDA) levels coupled with an inhibition of the superoxide dismutase (SOD) and catalase (CAT) activity. Co-treatment of cadmium and the antioxidants did alleviate the oxidative stress which was observed as a significant reduction in the MDA levels and an increase in the SOD activity both in the kidney and liver. CAT activity was only increased by vitamin C and Rosemary (15 mg/kg) co-treatment in the kidney. Furthermore, cadmium induced cellular disorganization of the kidney and liver was restored with the co-treatment of the antioxidants being more pronounced with the plant extracts. Rosemary leaf extract was efficacious at both the doses while the amla fruit extract had the most ameliorative effect at a higher dose (200 mg/kg).

Toxicological evaluations of methanolic extract of Moringa oleifera leaves in liver and kidney of male Wistar rats

This study was conducted to investigate toxicological effects associated with prolonged consumption of Moringa oleifera leaves as a beverage. Thirty rats were used in this study. They were grouped into five groups of six rats. Rats in group I received 2 mL/kg body weight (b.w.) of corn oil (vehicle). Animals in groups II, III, IV and V received 50, 100, 200 and 400 mg/kg b.w. of methanolic extract of M. oleifera (MEMO) for 8 weeks. Serum collected was analyzed for alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein, albumin, globulin, blood urea nitrogen (BUN) and creatinine. There was a significant (p<0.05) increase in serum total protein and globulin in a dose-dependent manner. Rats that received MEMO at 200 and 400 mg/kg b.w. showed a significant (p<0.05) increase in serum ALT, AST, BUN and creatinine which pointed to hepatic and kidney damage. All experimental animals that received MEMO had a significant (p<0.05) increase in body weight in a dose-dependent manner. This study therefore confirms for the first time that chronic administration of M. oleifera leaves might predispose to hepatic and kidney damage.

Three‐dimensional aspects of the structural characteristics and kidney angioarchitecture of adult and aged wistar rats: A scanning electron microscopy study

The aim of this study was to describe the structural morphological characteristics and organization of the kidney angioarchitecture in adult and aged Wistar rats using different scanning electron microscopy (SEM) methods. Twenty male Wistar rats (Rattus norvegicus) were divided into two groups: the Adult Group (n = 10), which included animals at 3 months of age, and the Aged Group (n = 10), which included animals at 24 months of age. Kidney samples from both groups were analyzed using three distinct SEM methods: coronal sections, cryofracturing and vascular corrosion casts/angioarchitecture. This study demonstrates the specificity of each technique for three-dimensional morphological analyses of the kidney using SEM. In the coronal sections, it was possible to view the structures on a given plane; the stratigraphy was best observed in the cryofracture method, and the vascular corrosion casts showed the organization and arrangement of the angioarchitecture. Thus, we concluded that the SEM methods together provide the best description of the morphological characteristics of normal and pathological kidneys of adult and aged Wistar rats.

Alterações histológicas dos rins e expressão das metalotioneínas e das proteínas de choque térmico em ratos Wistar após exposição ao fungicida tirame

Avaliaram-se as alterações histológicas e a expressão das metalotioneínas (MTs) e das proteínas de choque térmico (Hsp70) nos rins de ratos Wistar após a exposição ao fungicida tirame. Os animais foram distribuídos em três grupos: grupo dieta-padrão; grupo dieta-padrão+óleo de milho; e grupo tirame. Foram encontradas diferenças significativas (P&lt;0,05) na evolução do peso corporal entre os ratos do grupo tirame e os dos grupos controle e óleo de milho, e não foram verificadas lesões histológicas evidentes nos rins dos animais. Foram encontradas diferenças entre os animais do grupo exposto ao tirame e os dos grupos controle e óleo quanto às características histomorfométricas relativas ao corpúsculo renal - exceto para a proporção área da cápsula de Bowman:área do glomérulo - e relativas aos túbulos contorcidos proximal e distal - exceto para altura do epitélio dos túbulos distais. Nos ratos expostos ao tirame, foi observada imunomarcação positiva para as MTs, de moderada a forte, nos túbulos contorcidos da região cortical, diminuindo do córtex em direção à medula, e forte imunomarcação para as Hsp70 nas áreas do córtex e da medula, no glomérulo e nos túbulos contorcidos. Os resultados sugerem que o tirame pode ter toxicidade crônica nos mamíferos por afetar o seu crescimento e que a expressão das MTs e das Hsp70, provável resposta celular adaptativa ao estresse oxidativo causado pelo tirame, pode ser utilizada como biomarcador de exposição a este químico.

PROTECTIVE EFFECTS OF PROPOLIS ON GAMMA- IRRADIATED &lt;i&gt;NIGELLA SATIVA&lt;/i&gt; EXTRACT INDUCED BLOOD AND IMMUNE CHANGES IN WISTAR RATS

The present study conducted to test the effect of Nigella Sativa (NS, 5 mg kg -1 of body weight), or γirradiated Nigella Sativa (GRNS) on the changes of blood component profiles, liver, kidney functions and immune cytokines secretion in male Wistar rats. Moreover, the possible protection by propolis (200 mg kg -1 B. W.) on the changes induced by NS and GRNS was examined. Results revealed that both NS and GRNS administration for two weeks induced changes in blo od, GPT, GOT and urea levels and co-administration with propolis significantly ameliorated such change s. Also, liver histology showed numerous vacuolar degeneration and fatty changes in γ-irradiated groups which disappeared in presence of propolis. Kidney histology of NS administered rats showed less lymphocytic infiltration, while GRNS groups showed desquamation in the cytoplasm of the renal tubules, hemorrhage in the renal corpuscle and lymphocytic infiltration which disappeared when propolis given together with GRNS. Finally propolis induced protec tive effect on the changes induced in TNF-α and IL-10 secretion by either NS or GRNS in Wistar rats. In conclusion, the findings of present study clarified the protective effect of propolis on changes induc ed by γirradiated NS on blood, liver, kidney and cytokines changes in Wistar rats.

Effects of Aluminium Chloride Exposure on the Histology of lungs of Wistar Rats

Aluminium is ubiquitous element and the third most prevalent (abundant) element in the earth's crust. It is presents in many manufactured foods, medicines and is also added to drinking water for purification purposes. The purpose of this study was to evaluate the effects of aluminium chloride exposure on the histology of kidney of wistar rats. Twenty (20) adult wistar rats were used for this study. The animals were divided into five groups as follows: group I was the control that received distil water only, group II received 475mg kg -1 , group III received 950mgkg -1 , group IV received 1,425mg kg -1 and group V received 1,900mg kg -1 via oral intubation for duration of eight weeks. The wistar rats were humanely sacrificed and kidneys were removed and fixed in formalin. The kidneys were processed and stained using the routine haematoxylin and eosin staining method (H&E). The stained sections were examined under the light microscope fitted to a digital camera. Our results showed cavitations, glomerular atrophy and vacuolation of cytoplasm of the lining cells of both the proximal convoluted tubules (PCT) and distal convoluted tubules of the kidney in the treated groups when compared to the control group. Based on our observations, we therefore conclude that aluminium chloride exposure had detrimental effects on the histology of kidney of wistar rats.

Effects of trona on the redox status of cellular system of male rats

The effects of trona (Kaun), a food additive, on the redox status of the liver and kidney of male Wistar rats were investigated. A total of 60 male rats (145±2.52g) were grouped into four: A, B, C and D, where group A (the control) received 1mL of distilled water orally while those in groups B, C and D (test groups) received orally, same volume of trona preparation corresponding to 100, 200 and 400mg/kg body weight, respectively, for 28days. Administration of trona significantly reduced (p<0.05) alkaline phosphatase activity in the liver and kidney with corresponding increases in the serum enzyme. Acid phosphatase activity increased significantly (p<0.05) in the liver and kidney with no significant change (p>0.05) in the activity of the serum enzyme. The activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and the levels of reduced glutathione, vitamins C and E in the liver and kidney of the animals decreased significantly (p<0.05). In contrast, malondialdehyde and lipid hydroperoxide of trona-treated animals increased significantly (p<0.05) in liver and kidney. Overall, data from this study revealed that trona exhibited its toxic effect by suppressing or depleting the antioxidant systems and increasing the risk of attack by oxidants generated either from its metabolites or from other in vivo means on the rat cellular system.

Toxic effects of lead exposure in Wistar rats: Involvement of oxidative stress and the beneficial role of edible jute (Corchorus olitorius) leaves

Lead (Pb) is considered to be a multi-target toxicant. The present study was undertaken to evaluate the protective effect of aqueous extract of Corchorus olitorius leaves against Pb-acetate induced toxic manifestation in blood, liver, kidney, brain and heart of Wistar rats. The Pb-acetate (5mg/kg body weight) treated rats exhibited a significant inhibition of co-enzymes Q, antioxidant enzymes and reduced glutathione levels in the tissues. In addition, the extent of lipid peroxidation, DNA fragmentation and haematological parameters were significantly altered in the Pb-acetate treated rats as compared to control. Simultaneous administration of test extract (25, 50 and 100mg/kg body weight), could significantly restore the biochemical and haematological parameters near to the normal status through antioxidant activity and/or by preventing bioaccumulation of Pb within the tissues of experimental rats. Presence of substantial quantity of phenolics and flavonoids in the extract may be responsible for the observed protective role against Pb-intoxication.

Effects of vitamin E administration on phostoxin-induced changes in the kidney of adult Wistar rats

Aim: The effect of vitamin E on phostoxin-induced changes in biochemical parameters and the kidney of adult Wistar rats were studied. Materials and Methods: Thirty adult Wistar rats of both sexes were randomly separated into six groups of five rats each. Group 1 was the Control and was given normal saline. Group 2 was exposed to phostoxin for 3 hours per day with vitamin E and Group 3 was exposed to phostoxin for 1΍ hours with vitamin E. Groups 4 and 5 were treated with phostoxin only on exposure time of 1΍ and 3 hours respectively while Group 6 was treated with vitamin E only. The rats were exposed to phostoxin through inhalational method for 7 days and at the end of the exposure period, the rats were sacrificed. The blood and tissues were collected for analysis and were processed for histological studies. Results: The results showed significant changes in body weight of the rats (P < 0.05) while there was a significant increase in weight of the kidneys in Groups 3 and 6 when compared to the Control (P < 0.05). The results of the biochemical parameters Na + , K + , Cl−, urea, creatinine and HCO3 showed a significant difference (P < 0.05) between the control and the experimental groups. The results of histological examination of the kidney showed changes in animals exposed to phostoxin when compared to the Control animals. Conclusion: The changes in the kidney depend on the duration of the exposure time while vitamin E administration has been shown to have some level of protection on phostoxin-induced toxicity on the kidney tissues and biochemical parameters of the adult Wistar rats.

Toxicological effects of phosphide powder residue in female rats

Background: Fumigation of cowpea (Vigna unguiculata L. Walp.) using aluminium phosphide is a common practice and contact with this chemical has been demonstrated to result in organ damage, but there is dearth of data on the impact of its powder residue on a mammalian species. Aims & Objective: The aim of this study is to determine the effect of post-fumigation residue on both liver and kidney of female Wistar rats. Material and Methods: Eighteen rats were randomly divided into each of the three experimental groups. A group was fed with untreated cowpea and served as control; either of the two other groups was fed phosphide-powder residue contaminated or uncontaminated cowpea. Serum obtained from blood that was collected through retro-orbital bleeding was utilized for biochemical analysis. Results: While contact with treated but uncontaminated cowpea did not result in either hepatic or renal damage, rats fed phosphide-residue contaminated cowpea exhibited both hepatic and renal damage as the indices (alanine & aspartate aminotransferases, γ- glutamyl transferase, alkaline phosphatase, total protein, albumin, urea, uric acid, creatinine) of study were significantly different (p

Effect of aqueous leaves extract of Costus afer Ker Gawl (Zingiberaceae) on the liver and kidney of male albino Wistar rat.

Background:The use of medicinal plants in Nigeria has significantly increased over recent years as it is easily accessible, cheap and the strong belief that herbal remedies are natural and therefore non toxic.Aims:This study aims to investigate the sub-chronic toxicity (28-day) of the aqueous extract of Costus afer Ker Gawl leaves on the liver and kidney of male albino Wistar rats.Materials and Methods:A total of 20 male albino Wistar rats (113-205 g) divided into four groups of five weight-matched animals each, were used for the study. Group 1 received standard feed and water ad libitium and served as the control. Group 2, 3 and 4 received 375, 750 and 1125 mg/kg of aqueous extract of C. afer leaves respectively. The animals were sacrificed under ether anesthesia and the organs were harvested, weighed and histopathological studies carried out. The effect of C. afer on the hepatic biomarkers aspartate aminotransferase (AST), alanine aminotransferase (ALT); alkaline phosphatase (ALP); triglyceride (TG); total bilirubin (TB); conjugated bilirubin (CB); albumin (ALB) and kidney biomarkers urea, creatinine, sodium, potassium and bicarbonate were investigated.Statistical Analysis:Data were evaluated using Mann Whitney. If P ≤ 0.05 groups were considered to be significantly different.Results:C. afer contained alkaloids, saponins, flavonoids, anthraquinones, cardiac glycosides, terpenoids, phenolic compounds and tannins. The average body, organ, relative weights, feed and fluid intake showed no significant changes (P > 0.05) when compared to the control. The liver function tests (ALT, ALP, AST, CB, TB and ALB) showed significant differences (P < 0.05) in the test groups when compared with the control while TG showed no statistical difference (P > 0.05). The kidney function tests (urea, creatinine, sodium, potassium and bicarbonate) showed no significant differences (P > 0.05) in the test groups when compared to the control.Conclusion:Costus afer may be hepatotoxic but non-toxic to the kidney.

Mônica Piacentini Luizon1, Caroline Santos Oliveira1, Danielly Kathe Zaqueo1, Jaqueline Aparecida Oliveira1, Gustavo Ligieri Pereira do Prado2, Renata Barbieri1, Fernanda Oliveira Gaspari de Gaspi1, João Ernesto de Carvalho3, Elaine Cristina Mathias da Silva-Zacarin4, Camila Andréa Oliveira5, Acácio Antônio Pigoso1 and Grasiela Dias de Campos Severi-Aguiar5*

Oncidium flexuosum Sims. is an orchidwhose leaves are frequently used for therapeutic applications although its action mechanism is not exactly understood. So the aim of this study was investigate its effects in Wistar rat liver, kidney and bone marrow. For this purpose, twenty-five adult male Wistar rats were divided into five groups of 5 animals each: a control group receiving only water, a positive control receiving a mutagen compound and three groups treated for 15 days with 100, 250 or 500 mg/kg/day of a crude hydroalcoholic leaf extract of O. flexusosum Sims. No significant alterations were observed in –SH groups, MDA or catalase, showing that the extract does not induce hepatic oxidative damage. There was no evidence of kidney injury and hepatic function alteration (ALT). However, significant mitochondrial swelling was observed in the treated groups and is probably related to cell death induction observed in cells that displayed acidophilia, pyknosis and positive labeling with the TUNEL technique at higher concentrations of the extract. We demonstrated that the hepatotoxicity of the extract resulted from its ability to induce DNA and mitochondrial damage that arrests cells, leading to cell death, probably caused by the flavonoidsand tannins present in this extract.

Protective effects of vitamin C and selenium supplementation on methomyl-induced tissue oxidative stress in adult rats

Methomyl (MET) is used worldwide in agriculture and health programs. Besides its advantages in the agriculture, it causes several toxic effects. The objective of this study was to examine the potential ability of vitamin C and/or selenium (Se), to alleviate the oxidative damage parameters, against MET-induced changes in blood biochemical markers and oxidative damage in liver and kidney of male Wistar rats. The animals were randomized into five groups of eight each: group I served as control rats; group II received MET (8mg/kgbody weight (BW)) in drinking water; group III received both MET and vitamin C (200mg/kg BW; by intraperitoneal injection); group IV received both MET and Se (0.6mg/100gBW). Animals of group V were treated with MET, vitamin C and Se. A significant increase in the levels of hepatic markers enzymes (alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase) was determined. Furthermore, renal markers such as urea and creatinine were increased in MET-treated rats. Additionally, serum cholesterol and triglycerides were significantly enhanced. Exposure of rats to MET caused significant increase in malondialdehyde levels, thus causing a drastic alteration in antioxidant defense system, particularly in the activities of catalase and glutathione-S-transferase and glutathione peroxidase. However, simultaneous supplementation with vitamin C and Se restored these parameters partially. In conclusion, the results of the current study revealed that MET-induced toxicity caused perturbations of some biochemical parameters, lipid peroxidation and alterations in the antioxidant enzymes in liver and kidney homogenates. Administration of vitamin C and Se exhibited protective effect by inhibiting MET-induced toxicity in liver and kidney.

Effect of ischemia preconditioning on renal ischemia/reperfusion injury in rats

To study the effect of ischemia preconditioning (IP) on renal ischemia/reperfusion (I/R)-associated functional injury and expression of renal adhesion molecules in rats. The ischemia preconditioning plan adopted in this experiment involved renal warm ischemia for 6 min. and blood flow for 4 min., repeated four times. The Wistar rat kidneys used for warm ischemia preconditioning were subjected to 60 min of renal warm ischemia followed by reperfusion. The rat kidneys with ischemia/reperfusion were compared with the ischemia preconditioning group to observe rat renal function and changes in the expression of renal adhesion molecules ICAM-1, P-Selectin, and E-Selectin. The expression of rat renal adhesion molecules (ICAM-1, P-Selectin, and E-Selectin) with ischemia preconditioning was significantly lower than that of the ischemia/reperfusion group. Serum creatinine was significantly lower than that in the ischemia/reperfusion group after 48 hours. Ischemia preconditioning has a protective effect on renal function. Reduced expression of renal adhesion molecules is likely a mechanism involved in the observed protection.

Tissue damage in kidney, adrenal glands and diaphragm following extracorporeal shock wave lithotripsy

This study was designed to investigate whether exposure to short-term extracorporeal shock wave lithotripsy (ESWL) produces histologic changes or induces apoptosis in the kidney, adrenal glands or diaphragm muscle in rats. The effect of shock waves on the kidney of male Wistar rats (n = 12) was investigated in an experimental setting using a special ESWL device. Animals were killed at 72 h after the last ESWL, and the tissues were stained with an in situ Cell Death Detection Kit, Fluorescein. Microscopic examination was performed by fluorescent microscopy. Apoptotic cell deaths in the renal tissue were not observed in the control group under fluorescent microscopy. In the ESWL group, local apoptotic changes were observed in the kidney in the area where the shock wave was focused. The apoptotic cell deaths observed in the adrenal gland of the control group were similar to those observed in the ESWL groups, and apoptosis was occasionally observed around the capsular structure. Apoptotic cell deaths in the diaphragm muscle were infrequently observed in the control group. Apoptosis in the ESWL group was limited to the mesothelial cells. This study demonstrated that serious kidney, adrenal gland and diaphragm muscles damage occurred following ESWL, which necessitated the removal of the organ in the rat model. It is recognized that the ESWL complications related to the kidney, adrenal gland and diaphragm muscles are rare and may be managed conservatively.

Activating Nrf-2 Signaling Depresses Unilateral Ureteral Obstruction-Evoked Mitochondrial Stress-Related Autophagy, Apoptosis and Pyroptosis in Kidney

Exacerbated oxidative stress and inflammation may induce three types of programmed cell death, autophagy, apoptosis and pyroptosis in unilateral ureteral obstruction (UUO) kidney. Sulforaphane activating NF-E2-related nuclear factor erythroid-2 (Nrf-2) signaling may ameliorate UUO-induced renal damage. UUO was induced in the left kidney of female Wistar rats. The level of renal blood flow, cortical and medullary oxygen tension and reactive oxygen species (ROS) was evaluated. Fibrosis, ED-1 (macrophage/monocyte) infiltration, oxidative stress, autophagy, apoptosis and pyroptosis were evaluated by immunohistochemistry and Western blot in UUO kidneys. Effects of sulforaphane, an Nrf-2 activator, on Nrf-2- and mitochondrial stress-related proteins and renal injury were examined. UUO decreased renal blood flow and oxygen tension and increased renal ROS, 3-nitrotyrosine stain, ED-1 infiltration and fibrosis. Enhanced renal tubular Beclin-1 expression started at 4 h UUO and further enhanced at 3d UUO, whereas increased Atg-5-Atg12 and LC3-II expression were found at 3d UUO. Increased renal Bax/Bcl-2 ratio, caspase 3 and PARP fragments, apoptosis formation associated with increased caspase 1 and IL-1β expression for pyroptosis formation were started from 3d UUO. UUO reduced nuclear Nrf-2 translocation, increased cytosolic and inhibitory Nrf-2 expression, increased cytosolic Bax translocation to mitochondrial and enhanced mitochondrial Cytochrome c release into cytosol of the UUO kidneys. Sulforaphane significantly increased nuclear Nrf-2 translocation and decreased mitochondrial Bax translocation and Cytochrome c release into cytosol resulting in decreased renal injury. In conclusion, sulforaphane via activating Nrf-2 signaling preserved mitochondrial function and suppressed UUO-induced renal oxidative stress, inflammation, fibrosis, autophagy, apoptosis and pyroptosis.

Sulfur mustard causes oxidative stress and depletion of antioxidants in muscles, livers, and kidneys of Wistar rats

Sulfur mustard (SM) is a chemical warfare agent with cytotoxic effect and a tight link to oxidative stress (OS). Depletion of antioxidants is considered as a cause of detrimental consequence and belongs to the important steps leading to cell death. The oxidative injury appearing after SM exposure is not well understood. Nevertheless, identification of the pathological processes would be a good opportunity to establish an efficient therapy. Here, we focused our effort on an estimation of reactive oxygen species homeostasis and apoptotic processes in Wistar rats exposed to 0–160 mg/kg of SM. We assayed antioxidant activity, thiobarbituric acid reactive substances, reduced glutathione/oxidized glutathione, metallothionein, glutathione reductase, glutathione peroxidase, glutathione S-transferase, caspase 3, and glucose in the livers, kidneys, and muscles of the animals. Significant OS, depletion of low-molecular-mass antioxidants, increase in caspase activity, and some other processes related to SM action were determined. Moreover, we infer a principal role of OS in the tested organs.

Comparison of Fluconazole Renal Penetration Levels in Healthy and Candida albicans-Infected Wistar Rats

The aims of this study were to evaluate free levels of fluconazole (FCZ) in the kidneys of healthy and Candida albicans-infected Wistar rats using microdialysis and to establish the relationship between free renal and total plasma levels under both conditions. Microdialysis recovery rates were determined in vitro by dialysis, and retrodialysis recovery rates were determined in vivo by retrodialysis. The recovery rate was around 50%, independent of the method, drug concentration, or condition (in vitro or in vivo) used. FCZ kidney penetration in healthy and infected rats was investigated after the administration of 10 mg/kg of body weight intravenously (i.v.) or 50 mg/kg orally (n = 6/group) and blood and microdialysate sample harvesting at predetermined time points up to 24 and 18 h, respectively. There were no statistical differences between the area under the free concentration-time curve (AUC(0-∞)) values in plasma and in tissue for either healthy or infected groups for the same dose regimen investigated. The antifungal tissue penetrations were similar for both doses and under all conditions investigated (ranging from 0.77 to 0.84). The unbound fraction of FCZ was concentration independent (86.0% ± 2.0%), allowing the prediction of free renal levels using pharmacokinetic parameters obtained from total plasma fitting. The results showed that free renal and free plasma levels are similar in healthy and systemically C. albicans-infected rats. Therefore, free plasma levels are a good surrogate to estimate free FCZ renal concentrations in systemic candidiasis and can be used to optimize dosing regimens for this drug.

Development and Validation of a New GC-MS Method for the Detection of Tramadol, O-Desmethyltramadol, 6-Acetylmorphine and Morphine in Blood, Brain, Liver and Kidney of Wistar Rats Treated with the Combination of Heroin and Tramadol

Heroin is one of the most dangerous abused drugs in the world. Tramadol is an additive recently found at high concentration levels in street heroin seizures in Egypt. This substance could affect the usual analytical method for the detection of heroin and metabolites, as well as the pharmacokinetic and disposition of single analytes. One shortfall regarding this issue is present in the literature. This study describes a validated, simple, sensitive and selective method to determine tramadol, O-desmethyltramadol, 6-acetylmorphine and free morphine in the blood, brain, liver and kidney of Wistar rats, intraperitoneally treated with a combination of heroin and tramadol (10 and 70 mg/kg, respectively) using liquid-liquid extraction and gas chromatography-mass spectrometry detection. The calibration curves of tramadol, O-desmethyltramadol and 6-acetylmorphine in blood were linear in the concentration range from 25-5,000 ng/mL and morphine was found in the concentration range 50-5,000 ng/mL. The analytes were detected in all tested matrices, except 6-acetylmorphine, which was not detected in liver. The highest concentrations of tramadol and O-desmethyltramadol were observed in kidney (22,9381 and 28,498 ng/g), while 6-acetylmorphine and morphine were found at the highest levels in brain (3,280 and 3,899 ng/g, respectively). The present method is simple, rapid and sensitive and can be used to study the pharmacokinetics, disposition and interaction of these drugs in several animal models.