Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P.

109 – 117

Nigerian Journal of Pharmaceutical Sciences

Vol. 8, No. 1, March, 2009, ISSN: 0189-823X
All Rights Reserved

MICROBIOLOGICAL QUALITY OF SOME COMMERCIALLY

AVAILABLE PAEDIATRIC ANTI-MALARIAL AND COUGH
PREPARATIONS IN ILORIN, NIGERIA

Adeshina, G. O., Ajayi, S., Onaolapo, J. A.

1
Department of Pharmaceutics and Pharmaceutical Microbiology, Ahmadu Bello University, Zaria,
Nigeria

*Author for Correspondence: dotunkele@yahoo.com, 08037880000

ABSTRACT
The bacteriological quality of twenty (20) different brands of paediatric anti-malaria and cough
preparations sold in Ilorin, Nigeria was investigated. The bacterial load was determined using the
viable cell count method. The bacteria isolated were characterized by standard methods. The level of
contamination in fourteen (14) preparations exceeded the official tolerance limit of permissible
microorganisms specified for syrups and suspensions. Some of the preparations were contaminated
with Bacillus subtilis, Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa.
Bacillus subtilis was found to be the most predominant contaminants. This study has shown that some
paediatric anti-malarial and cough preparations sold in retail outlets in Ilorin, Nigeria are heavily
contaminated with microbial agents.

Key words: Bacteriological, quality, Syrups, Suspensions

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Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117
INTRODUCTION
Malaria is one of the major killer
diseases of the world. Malaria causes up
to 2.7million deaths per year with the
vast majority of these among young
children in Africa especially in remote
rural areas with limited or no access to
medical care. In some parts of Africa,
malaria kills 3000 children less than 5
years of age each day (Rang and Dale,
2005). Cough is common among infants
at various stages of development. At the
teething stage, because of the itching
sensation produced by the gums they
pick any material ranging from toys to
other objects to scratch the gum and
relief such feelings; this can stimulate
cough as some of the objects are already
contaminated with microbes which can
cause inflammation and then infection of
the respiratory tract. Cough is also a
common ailment during some specific
times of the year especially among
children (Rang and Dale, 2005).
Syrups and suspensions which are
basically prepared for children are non-
sterile liquid pharmaceutical dosage
forms. They are prepared for oral
administration in children since tablets
and capsules cannot be easily or
conveniently administered to them.
Syrups are aqueous preparations
characterized by sweet taste and a
viscous consistency while suspensions
are disperse systems; they are both used
as vehicles to convey anti-malarial and
cough medications to ensure palatability
and easy administration.
The total viable count of permissible
bacteria (i.e. non-pathogenic bacteria)
must not exceed 103cfu/ml while yeast
and mould should not exceed 102cfu/ml.
Furthermore liquid pharmaceutical
preparations are to be devoid of E.coli
and other coliforms; Pseudomonas
aeruginosa, Staphylococcus aureus and
110
other pathogens in accordance with the
Reports of Mixed Working Committee of
Official Laboratory and Drug Control
Services and a section of the International
Pharmaceutical Federation (1975). Some
other organisms such as Bacillus subtilis
which were regarded as non pathogenic
have been reported to cause severe
infection in human (Cotton et al., 1987;
Sliman et al., 1987). Bacterial
contamination of syrups and suspensions
medications can cause spoilage of the
product and constitute direct health
hazard to the patients due to the
metabolic versatility of bacteria in which
almost any formulation ingredient, from
simple sugars to complex aromatic
molecules may undergo chemical
modification by a suitable organism
(Takon and Antai, 2006). There has been
increasing number of reports of infections
caused by contaminated non-sterile
preparations (Ibezim, 2002). The extent
of microbial contamination depends on a
number of factors such as availability of
nutrients, presence of microorganisms,
and oxygen e.t.c. and also the factors
determining the outcome of medicament-
borne infection include the type and
degree of microbial contamination, route
of administration and the state of the
patient’s immune system (Baird, 2004).
There is the need for Good
Manufacturing Practice (GMP) in the
production of non-sterile preparations
like syrups and suspensions, which are
specifically meant for children, to ensure
that the products are consistently
manufactured to a quality appropriate for
its intended use. The need for Good
Manufacturing Practice cannot be
undermined to generally decrease
microbial load of these products so as to
prevent the children being treated from
developing infections which can be
severe due to contamination of these
Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117

medicaments with pathogenic MATERIALS AND METHODS

microorganisms.
The present study was designed to
enumerate the microbial contaminants
present in some brands of paediatric
cough and anti-malarial preparations
marketed in Nigeria and to compare with
the permissible level of aerobic bacteria
contaminants officially allowed.
A total of twenty (20); ten (10) anti-
malaria and ten (10) cough paediatric
preparations which were purchased
from Pharmacy shops in Ilorin, Nigeria
were used in the study. Five samples
of each brand were purchased from
different Pharmacy shops in Ilorin
metropolis, Nigeria.

Table 1: Paediatric anti-malarial preparations investigated

Samples Constituents Dosage Manufacturing Expiring date

form date
A1 SFD + PRM 08/08 11/11
Suspension
A2 AMQ 05/07 04/10
Suspension
A3 ATM + LMF 07/07 06/09
Suspension
A4 ATS 07/07 07/10
Suspension
A5 SFD + PRM 05/08 05/10
Suspension
A6 SFD + PRM 01/08 12/09
Suspension
A7 CLQS Syrup 05/08 04/11
A8 CLQ Syrup 01/08 04/10
A9 CLQP Syrup 02/08 02/11
A10 CLQP+PMT+PCM Syrup 06/08 05/10
Key: SFD + PRM = Sulfadoxine + Pyrimethamine
AMQ = Amodiaquine Hydrochloride
ATM + LMF = Artemether + Lumefantrine
ATS = Artesunate
CLQS = Chloroquine Sulphate
CLQ = Chloroquine
CLQP = Chloquine Phosphate
PMT + PCM = Promethamine + Paracetamol

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Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117

Table 2: Paediatric cough syrups investigated.

Samples Constituents Manufacturing Expiring date

date
B1 DPH + MTL 05/08 05/11
B2 DPH B.P+SCT B.P 07/08 06/11
B3 DPH + SCT Ph Eur 06/08 06/11
B4 DPH PhEur + 06/08 11/10
AMC+TSC+CA+MTL+FSP
B5 PMZ + PCD + ETL 11/07 10/10

B6 DPH+SCT+AMC+MTL 04/08 03/11

B7 DPH Ph Eur 08/07 08/10
B8 DPH + SCT 09/07 09/09
B9 TPD B.P+PED B.P+CDP 12/08 12/09
B.P + PGS
B10 TPD B.P + PED B.P + CDP 06/08 06/10
B.P + PGS
Key: DPH = Diphenhydramine HCl, MTL = Menthol, SCT = Sodium citrate B.P,
AMC = Ammonium chloride, TSC = Trisodium citrate, CA = Citric acid,
FSP = Flavoured syrup, PMZ = Promethazine, PCD = Pholcodine
TPD = Tripolidine HCl, PED = Pseudoephedrine HCl B.P,
CDP = Codeine phosphate B.P, PGS = Potassium Gualacolsulphate

Enumeration and Isolation of units per ml of each syrup and

Microbial Contaminants suspension calculated (Conn, 1975).

Using the spread plate technique, each Identification of Isolated

sample of the syrup was vigorously
shaken; 0.2ml was withdrawn and
diluted serially (10 fold dilution) in
sterile normal saline. A quantity of 0.1ml
of the diluted sample was spread on the
surface of nutrient agar or Sabouraud’s
Dextrose agar (SDA) plates. The
suspensions were reconstituted with
sterile distilled water and were
thoroughly mixed and cultured as for the
syrups. The nutrient agar plates were
incubated at 37oC for 24hours while the
SDA plates were incubated at room
temperature for 3 days. All experiments
were done in duplicates and controls set
up in each round. Colonies were counted
and the mean number of colony forming
Microorganisms
The sample of the syrups and
suspensions were plated on various
selective media such as MacConkey agar
(for enteric bacteria), Cetrimide agar
(Pseudomonas), Mannitol Salt agar (
Staphylococcus) and Sabouraud’s
Dextrose Agar ( mould and yeasts).
Growth characteristics, microscopy
study and biochemical tests were used
for the identification of isolates (Cowan,
1974; Collee and Miles, 1989).

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Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117
RESULTS
All the preparations tested were
contaminated with bacteria but none was
contaminated with yeast or mould. The
results of the total viable count of
bacteria in the samples are as shown in
Tables 1 and 2. The bacterial count of
the tested anti-malarial preparations
ranged from 6.00 x 102cfu/ml to 2.70 x
106cfu/ml. The highest viable count was
recorded in Chloroquine phosphate +
Promethamine HCl + Paracetamol B.P
(A10) with a count of 2.70 x 106cfu/ml
while sample A7 (Chloroquine sulphate)
had the lowest bacterial count of 6.00 x
102cfu/ml (Table 1).
In the group of cough syrups, the
bacterial count ranged from 2.50 x
102cfu/ml – 4.20 x 105cfu/ml. Sample
B5 (Promethazine HCl B.P + Pholcodine
B.P) had the highest bacteria count of
4.20 x 105cfu/ml while B3 (
Diphenhydramine HCl Ph Eur + Sodium
citrate Ph Eur) had the lowest bacterial
count of 2.50 x 102cfu/ml (Table 2).
Table 3: Viable bacterial counts in the paediatric anti-malarial suspensions
and syrups
Sample
A1
A2
A3
A4
A5
A6
A7
A8
A9
A10
Six samples A7, B1, B2, B3, B4, B5,
were contaminated to a level permitted
officially i.e less than 103 cfu/ml aerobic
bacterial counts. All the suspensions
investigated were contaminated above
the permissible limit for non-sterile
pharmaceutical preparations (Tables 1
and 2).
The distribution pattern of isolated
contaminants in the anti-malarial and
cough suspensions and syrups are as
shown in Table 3 and 4. Bacillus subtilis
was isolated from all the samples of the
syrups and suspensions constituting the
greatest contaminants of the products.
E.coli was isolated from samples B3,
B4, and B9 which were all cough syrups.
Two of the anti-malarial suspensions
(A1 and A4) were contaminated with
Staphylococcus aureus. Only one of the
samples A2 (anti-malarial suspension)
was contaminated with Ps. aeruginosa.
No mould/yeast was isolated from any of
the syrups or suspensions.
Dosage form Bacterial count (cfu/ml)
Suspension 1.39 x 105
Suspension 1.10 x 103
Suspension 1.80 x 105
Suspension 5.10 x 103
Suspension 6.00 x 105
Suspension 2.43 x 106
Syrup 6.00 x 102
Syrup 5.00 x 105
Syrup 1.75 x 104
Syrup 2.70 x 106
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Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117

Table 4: Viable bacterial counts in the paediatric cough syrups

Sample Dosage form Bacterial count (cfu/ml)

B1 Syrup 9.90 x 102
B2 ” 9.00 x 102
B3 “ 2.50 x102
B4 “ 7.00 x 102
B5 “ 4.20 x 105
B6 “ 5.50 x 102
B7 “ 6.00 x 103
B8 “ 3.15 x 105
B9 “ 1.00 x 105
B10 “ 2.65 x 104

Table 5: Distribution of bacterial contaminants in the paediatric anti-malaria

Preparations
Sample Dosage form Bacillus E .coli Staph. Ps.
subtilis aureus aeruginosa
A1 Suspension + - + -
A2 Suspension + - - +
A3 Suspension + - - -
A4 Suspension + - + -
A5 Suspension + - - -
A6 Suspension + - - -
A7 Syrup + - - -
A8 Syrup + - - -
A9 Syrup + - - -
A10 Syrup + - - -
Key: + Present
- Absent
Table 6: Distribution of bacterial contaminants in the paediatric cough syrups

Sample Dosage form Bacillus E .coli Staph. Ps.

subtilis aureus aeruginosa
B1 Syrup + - - -
B2 Syrup + - - -
B3 Syrup + + - -
B4 Syrup + + - -
B5 Syrup + - - -
B6 Syrup + - - -
B7 Syrup + - - -
B8 Syrup + - - -
B9 Syrup + + - -
B10 Syrup + - - -
Key: + Present; - Absent

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Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117
DISCUSSION
Only six brands out of the twenty brands
of paediatric preparations tested A7, B1,
B2, B3, B4 and B6 had bacterial load of
less than 103cfu/ml which implies that
they complied with the official
requirement for the microbiological
quality of syrups and suspensions
according to the FIP Working
Committee (1975).
Four (4) groups of bacteria; Bacillus
subtilis, Escherichia coli, Pseudomonas
aeruginosa and Staphylococcus aureus
were isolated from the samples. The
types of bacterial contaminants isolated
suggest the route of contaminations
possibly water, personnel and
environment. These bacterial species
have previously been associated with
drug contaminations (Mendie and
Hugbo, 1993; Ibezim et al, 2002; Takon
and Antai, 2006).
Fourteen (14) brands; five (5) cough and
nine (9) anti-malarial preparations were
heavily contaminated and thus did not
meet the official limit. This
contamination can be attributed to poor
manufacturing practice; bacterial
contaminants could be from the raw
materials, the air in the manufacturing
environment, water used or the
personnel involved, packaging process
or containers and equipments (Baird,
2004). It has been reported that most
drugs were contaminated by
microorganisms during handling and
storage (Tankon and Antai, 2006).
Suspensions were slightly more
contaminated than the syrups. Syrups
contain very high content of sucrose
(65%) which inhibits the growth of
bacteria and moreover syrups are usually
filtered prior to bottling, while
suspensions contain natural products as
115
suspending agents which can encourage
heavy bacterial contamination.
Anti-malaria preparations such as A1,
A5 and A6 contained sulfadoxine, a
sulphonamide; A8, A9 and A10
contained Chloroquine which should
inhibit bacterial growth, regardless of
this fact; they were contaminated above
the permissible limit. Contamination of
products may affect their stability
causing product degradation prior to
expiration date (Adeshina, 2008) and
this can also lead to infections especially
in this case where children, whose
immunological system is weak, are
involved.
From the results, all the tested samples
were contaminated with B. subtilis.
Some recent studies have shown that
Bacillus was the most frequent
contaminant of non-sterile
pharmaceuticals (Maria et al., 1993;
Ibezim et al., 2002). Members of this
genus are widespread in the air, soil,
water and in animal products such as
hair, wool and carcasses (Willey et al.,
2008).
The presence of E.coli in some of the
samples indicated faecal contamination
which may be principally from
production personnel and possibly from
the water used as vehicle. Escherichia
coli are not always confined to the
intestine and their ability to survive for
brief periods outside of the body makes
them an ideal indicator organism to test
samples for faecal contamination
(Willey et al., 2008). There was a
reported incidence of infant diarrhea due
to E.coli in some parts of Nigeria which
resulted from the quality of water supply
(Antai and Anozie, 1987).
Pseudomonas aeruginosa, which is a
recalcitrant drug contaminant, and Staph.
aureus were isolated from samples A4,
A1 and A4 respectively. It has been
Adeshina et al., Nig. Journ. Pharm. Sci., March, 2009, Vol. 8 No. 1, P. 109 – 117
reported that most liquid drugs were
commonly contaminated by Ps.
aeruginosa (Mendie and Hugbo, 1993).
The Pseudomonas contamination of this
sample is of great public health
significance, as Ps. aeruginosa isolated
from environment often present
resistance to clinically available anti-
microbial agents. Also Pseudomonas
spp., isolated from some bottled water
and orange drinks were resistant to one
or more antibiotics (Kailis et al., 1991;
Olutimayin and Onaolapo, 1997). Some
Staphylococcus aureus strains isolated
from human wounds have been shown to
be resistant to some antibiotics
(Olayinka et al, 2003).
The major sources of contamination of
pharmaceuticals have always been
water, the production environment, the
personnel and packaging material
(Olutimayin and Onaolapo, 1997).
Therefore proper attention should be
given to the prior treatment of these
CONCLUSION
The results of the microbiological study
of twenty brands of paediatric anti-
malaria and cough preparations
marketed in Ilorin revealed that fourteen
of the twenty preparations tested were
highly contaminated above the official
permissible limits of microbial load of
non-sterile pharmaceutical preparations
(syrups and suspensions). It is therefore
suggested that Good Manufacturing and
packaging Practice, proper treatment of
water and air; personal hygiene
improvement of the production
personnel and pretreatment of natural
raw materials be enforced and
maintained. Also proper handling and
storage of these products be carried out
to eliminate or reduce microbial
116
factors to ensure reduction in the level of
microbial contaminants. The
incorporation of sufficient concentration
of appropriate preservatives can also be
employed to reduce the microbial load of
these preparations. Good Manufacturing
Practice can therefore not be overlooked
in pharmaceutical industries whether
sterile or non-sterile preparations as the
health of the patients, in the case of the
little child are of paramount importance.
Drugs used in the treatment of prevalent
diseases among children like malaria and
cough cannot afford to be heavily
contaminated, since those formulated as
powders (suspensions) are reconstituted
with water that the quality cannot be
guaranteed especially in areas where
access to clean portable water is scarce
and children who take this medications
have weakened immunological system
already which makes them vulnerable to
infections.
contamination to meet the official
requirement.
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