Evaluation of Acute and Chronic Toxicity of Tartrazine (E102) on Steriod Reproductive Hormones of Albino Rats

Abstract

Aim: To determine the acute and chronic effect of tartrazine on reproductive steroid hormones of albino rats. Study Design: The design involved acute and chronic study. The acute study investigated intraperitoneal and oral route of administration while the chronic study used oral route only. The rats used for the study weighed 150 gm approximately. In the acute study, 48 rats (24 female and 24 male) were used for intraperitoneal treatment and were randomly selected into six groups treated with 0.0 g/kg, 1.67 g/kg, 3.33 g/kg, 5.0 g/kg, 6.67 g/kg and 8.33 g/kg of tartrazine. In orally treated rats, 48 rats (24 female and 24 male) were also used and were treated with 0.0 g/kg, 2.5 g/kg, 5.0 g/kg, 10.0 g/kg, 15.0 g/kg and 20.0 g/kg of tartrazine. In the chronic study, the experiment was divided into phase 1, 2 and 3 which lasted for 30, 60 and 90 days respectively. In each phase, 80 rats were used and were divided into treatment and control groups. The treatment groups were given 7.5 mg/kg of tartrazine orally on daily basis over a period of 30, 60 and 90 days while the control groups were not treated with tartrazine. Place and Duration of Study: The study was carried out in the Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria over a period of 12 months (December, 2017 – December, 2018). Methodology: At the end of the acute and chronic study, 5 mls of whole blood specimens was collected by means of cardiac puncture into plain bottles. The specimens were spun at 4500 rpm for 10 minutes to obtain serum. The laboratory analysis of the hormonal parameters was based on Enzyme Linked Immunosorbent Assay (ELISA) Technique. Statistical analysis was performed using GraphPad Prism version 5.03. More so, ovarian and testicular tissues were also collected for histological examinations. These tissues were fixed in 10% formol-saline prior to tissue processing. Staining was done using Haematoxylin and Eosin stain. Results: In acute study, female treated rats (intraperitoneally and orally) showed significantly higher values in Progesterone (PROG) and Estradiol (E2) concentrations while male treated rats (intraperitoneally and orally) indicated significantly lower values in testosterone (TESTO) concentration compared with control rats. Histopathologic examination showed flagella distortion in the seminiferous lumen, vacuolation, pycnosis, distortion of basement membrane and loss of leydig cells of the testis. More so, mild vacuolation of follicular ovarian cells were also seen. In chronic treatments, hormonal parameters after 30 days, 60 days and 90 days showed no significant differences in testosterone (TESTO), Progesterone (PROG) and Estradiol (E2) concentrations in tartrazine treated rats compared with their respective control rats. When the comparative analyses of treated groups after 30, 60 and 90 days using One-Way ANOVA were considered, testosterone (TESTO) concentration indicated significantly lower levels in treated male rats while Progesterone (PROG) showed significantly higher values over 30, 60 and 90 days in treated female rats. Histopathologic examination indicated mild changes such as flagella distortion, pycnosis and vacuolation in testicular tissues especially after 90 days of chronic treatment likewise mild vacuolation of ovarian cells. Conclusion: In the acute study, reduction in testosterone (TESTO) concentration while increase in PROG and E2 concentrations were seen. However, in the chronic study, significant differences were not seen in testosterone (TESTO), Progesterone (PROG) and Estradiol (E2) concentrations. Finally, when the influence on duration of exposure at ADI doses (7.5 mg/kg) were considered after 30, 60 and 90 days, reduction in testosterone (TESTO) and increase in Progesterone (PROG) concentrations were seen.