The Effect of Sub-acute Administration of Cough Syrup with Codeine on Biomarkers of Oxidative Stress in Adult Female Wistar Rats


Akor-Dewu M. B., Wada, H.U.,and Abdulkareem,J.
Department of Human Physiology, Faculty of Basic Medical Sciences,  College of Health Sciences, Ahmadu Bello University, Zaria, Kaduna State, Nigeria.

Department of Human Physiology, Faculty of Basic Medical Sciences,College of Medicine, Kaduna State University, Kaduna State, Nigeria.

All correspondence to: E-mail:;


The aim of this study is to investigate the effects of cough syrup containing codeine on lipid peroxidation, some oxidative stress biomarkers (catalase (CAT) activity, superoxide dismutase (SOD) activity, glutathione (GSH) concentration) and haematological indices (red blood cell count(RBC), white blood cell count (WBC), packed cell volume (PCV) and haemoglobin concentration (Hb)) in female Wistar rats. Twenty female Wistar rats were grouped into four groups, with five animals in each group. Group 1 (control) received 10ml of normal saline/kg body weight, whilegroups 2, 3 and 4 received 10.9 5 mg/kg, 21.90 mg/kg and 43.80mg/kg Benylin® with codeine respectively for 14 days, all administrations were given via oral gavage.After 14 days of administration the animals were sacrificed, 5mls of blood was collected by cardiac puncture and serum collected by centrifugation wasanalysed for antioxidant enzymes by colorimetric enzymatic methods. Whole-blood collected was used to manually analyse for some haematological parameters. The result showed no significant difference (p > 0.05) in MDA, SOD, GSH, RBC, WBC, PCV and Hb concentration between the groups, but catalase activity was significantly (p < 0.05) increased in group 3 (8.028±0.213u/mg) and group 4 (7.668±0.390u/mg) when compared to control (3.834±0.750u/mg). In conclusion, increase in catalase activity was observed at higher concentrations (21.9mg/kg and 43.8mg/kg) of Benylin® with codeine.

Keywords: oxidative stress, haematology, cough syrup, codeine.


Drug misuse refers to the use of drugsor prescription medicationfor alternative purposes from its original prescription, stimulating effects such as mood alteration or intoxication (Powell, 2011). Drug abuse may be defined as the “arbitrary” overdependence or misuse of drugs with or without a prior medical diagnosis from qualified health practitioners (Lakhanpal and Agnihotri, 2007).  Substances of abuse are categorized as: stimulants, hallucinogens, narcotics, depressants (sedatives and tranquilizers) cannabis, inhalants and anabolic steroids (Bassiet al., 2017).

The most commonly abused cough syrups are those containing Codeine. Codeine is an opiate used for its analgesic, antitussive and antidiarrheal properties, acting centrally to elicit its pharmacological effects (Uthmanet al., 2016). Despite its clinical benefits, dependence on codeine when it is abused or misused is a potential problem,according to Lawan and Adie (2012),“the abuse of cough syrups in Nigeria,especially among women of youthful and adult age is quietlyspreading like wild fire.” Addiction to codeine syrup is turning the otherwise conservative girls and women of the north into social miscreants and rebellious housewives. It is increasingly becoming common to see girls and women at ceremonies and social activities where they drink codeine allowing them to forget about all inhibitions.Women mix these drugs in drinks during ceremonies and share it among themselves, the goal is to get tipsy and have courage to dance and talk vulgarly. The major drug abused by girls and women is cough syrup containing codeine, which they take in large quantities to make them tipsy and almost impervious to pain, just the way narcotics do (Lawan and Adie, 2012; Uthmanet al., 2017).The potential for overuse and misuse of codeine containing medications is not only detrimental to a person’s health but has economic and social implications (Feinberg, 2006).

The misuse, abuse and dependence on codeine products present in a range of over the counter medicines that are dispensed to the public without prescription in now public health challenge throughout the world (Cooper, 2013).Therefore the aim of this study is to investigate the effects of cough syrup containing codeine on lipid peroxidation, some oxidative stress biomarkers and haematological indices. 


Twenty (20) female Wistar rats weighing 150-200g were purchased from the National Veterinary Research Institute, Vom, Jos, PlateauState. The animals were housed in the Animal House of the Departmentof Human Physiology,

Faculty of Medicine, Ahmadu Bello University, Zaria and The Effect of Sub-acute Administration of Cough…

were acclimatized for 2 weeks before the commencement of the experiment. The animals weremaintained on standard diet (vital feeds) and water. Experiments were carried out between 9:00am and 12:00 pm daily in accordance with the Guide for the Care and Use of Laboratory Animals as adopted by the National Institutes of Health.Weighing of the animals were done continuously each day before administration of the drug.Commercial Benylin® with Codeine was donatedfor this research from a licenced Pharmacy in Zaria, Kaduna State Nigeria.

Experimental Design

The animals were divided into four groups each weighing 150 ± 50 gram. Their weights were taken every day for the period of two (2) weeks before administration. The drug were administered orally to the rats by carefully inserting the cannula into the oral cavity of the rats. Animals of group 1 were treated with 1ml/kg normal saline which served as the control, while animals in group 2,3 and 4 received single daily dose of 10.95mg/kg, 21.90mg/kg and 43.8mg/kg of Benylin® with Codeine Syrup adopted from Tijjaniet al.,(2012). At the last day of administration, the animals were sacrificed using ketamine hydrochloride and blood samples (5mls) were collected from each subjectby cardiac puncture. An average of2.5ml of each blood sample wasstored in plain tubes for 20 minutes, then centrifuged to collect the serum which was usedfor evaluation of biomarkers oxidative stress.The remaining and 2.5mls of each blood sample was stored in EDTA bottles for RBC, WBC, PCV and Hb estimation.


Biomarkers of oxidative stress were evaluated by colorimetric enzymatic methods as follows:

Malondialdehyde by Okhawaet al.(1979); catalase activity by Aebiet al. (1974); superoxide dismutase activity by Maetinet al. (1987); glutathione concentration by Rajagopalan et al. (2004). Haematological parameters were measured by methods described by Lewis et al.(2006).

Statistical Analysis

Data obtained were processed and One Way analysis of variance (ANOVA) was used to check for significant differences. Tukey’s post-hoc test was implored to determine the level of significance. Results are expressed as mean ± SEM. Values of p<0.05 were considered significant.Graphpad prism 6.was used for the analysis


There was a significant increase for catalase concentration in group 3 and 4 (table 1) when compared to the control, while SOD, MDA, GSH and TP showed no significant difference in their concentration between the groups. There was no significant difference (table 2) in blood parameters investigated.

Table 1: Levelsof Superoxide dismutase (SOD), Malonyldehyde (MDA), Catalase(CAT), Glutathione(GSH) and total protein content (TP) in the female Wistar rats administered with Benylin® containing codeine.

Experimental      group SOD

(u/mg protein)


(u/mg protein)


(nmol/mg protein)


(nmol/mg protein)




(group 1)

Group 2

(10.95 mg/kg) Group 3

(21.90 mg/kg) Group 4

(43.80 mg/kg)





















* is significant at p < 0.05, CAT = catalase, = superoxide dismutase, GSH = glutathione,    MDA= malondialdehyde, TP = total protein

Table 2. Haematological parameters of the blood of female Wistar rats administered with Benylin® containing codeine.

  RBC (x1012 L) WBC (x109L) PCV(%) Hb(g/dL)



(10.95 mg/kg)


(21.90 mg/kg)


(43.80 mg/kg)

3.97 ± 0.17

3.96 ± 0.14

3.99 ±0.12

4.01± 0.20

1.31 ± 0.95

1.10 ± 0.61

1.17± 0.93

1.25± 0.64

43.0 ± 0.95

43.8 ± 1.02

44.3± 0.48

42.4 ±1.03

14.2 ± 0.42

14.6 ± 0.34

14.9 ±0.30

14.1± 0.34

RBC = red blood cell count, WBC = white blood cell count, PCV = packed cell volume, 

Hb = haemoglobin concentration



Results obtained from the present study showed no significant difference in the level of activities of MDA,SOD and GSH when the experimental groups were compared with control. This may indicate that the antioxidant enzymes MDA,SOD and GSH may not be affected by codeine administration in rats, in this study. However, a significant increase was observed in catalase activity, and this may suggest that catalase may be a more sensitive antioxidant enzyme when codeine is being administered. Again, the finding may alsopropose that catalase is a better tool to monitor in patients administered with codeine.Chattopadhyay et al.,(2007) reported that an increase in some antioxidant enzymes activities such as SOD and CAT, may be indicative of the failure of compensation in oxidative stress. These enzymes scavenge excess O2, H2O2, and superoxide anion radical to H2O2, the resultinghydrogen peroxide in turn is decomposed by the enzymes Gpx and CAT leading to functional changes induced by radical over- production and an increase in the biosynthesis of antioxidant enzymes (Vendittiet al., 2003). There was no significance in haematological parameters when compared to control but a sedating and calming effect was noted in the rats during the period of administration. This behaviour can be compared to that of Tijjaniet al., (2012) where they reported decreased activity

(hypoactivity) in mice treated with single doses of

21.90mg/kg and 43.80mg/kg of Benylin® with Codeine.Sacerdoteet al. (1997) reported that codeine possesses a weak antinociceptive effect and limited immunosuppressive activity, this could reflect the result of no significance in haematological parameters

The insignificant change in haematological parameters caused by Benylin with Codeine suggests that the immune system might not have been compromised, eventhough other immune responses were investigated.


Higher concentrations of Benylin with Codeine administration elicited a rise in the activity of catalase, but not with malondialdehyde, superoxide dismutase, or glutathione in rats. In addition, values of RBC, WBC, PCV and Hb were not significantlyaffected by administration of Benylin with Codeine in this study.


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