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GLUTEN SCREENING FROM
PHARMACEUTICAL PRODUCTS, AN
IMPERATIVE FOR CELIAC DISEASE
MANAGEMENT
SIMONA OANCEA1*, MIHAELA STOIA2
University “Lucian Blaga” of Sibiu, Department of Biochemistry and
Toxicology, Str. I. Raţiu 7-9, 550012 Sibiu, Romania
2
Authority of Public Health, Sibiu, Str. Gh. Bariţiu 3, Sibiu, Romania
*corresponding author: e-mail: simona.oancea@ulbsibiu.ro
1
Abstract
Celiac disease is a chronic intestinal disorder caused by intolerance to gluten. It
is recognized that pharmaceutical products may contain traces of gluten which will cause
acute illness in celiac patients. In the present paper we investigated the content of
potentially hidden gluten ingredient in basic children drugs using the
immunochromatographic visual test. The negative gluten results are relevant to complete
the existing lists of gluten-free drugs. The visual assay proved to be an efficient rapid tool
for gluten screening as an alternative to the enzyme-linked immunosorbent assay (ELISA)
techniques.
Rezumat
Boala celiacă reprezintă o disfuncţie intestinală cronică, rezultat al intoleranţei la
gluten. Astăzi este recunoscut faptul că numeroase produse farmaceutice pot conţine urme
de gluten ce determină simptome acute la pacienţii cu boala celiacă. În articolul prezent, am
investigat conţinutul de gluten ca potenţial ingredient în produsele farmaceutice de bază
destinate copiilor, utilizând metoda vizuală imunocromatografică. Rezultatele negative
obţinute sunt importante şi relevante pentru completarea listelor existente, cu privire la
medicamentele lipsite de gluten. Analiza imunocromatografică s-a dovedit o metodă
eficientă şi rapidă de screening al glutenului ca alternativă la metodele ELISA.
Keywords: celiac disease; gluten; immunochromatographic test
Introduction
Data from the National Institute of Health (NIH) reported that 1%
of USA population is suffering from celiac disease (CD). In Europe, 1 in
150-300 individuals may suffer from CD, which still remains
underdiagnosed in many European countries [1]. CD is a food-induced
disorder characterized by a permanent intolerance to wheat gluten and
related proteins from rye and barley. The ethiology of CD is caused by
digestive disorders of the intestinal mucosa. The pathogenesis of the disease
develops under an immunological mechanism driven by human leukocyte
antigens-(HLA-DQ2 or DQ8)-restricted T cells [2]. Clinically, the picture of
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CD goes from typical symptoms of diarrhea, abdominal pain, weight loss
and anaemia to atypical symptoms of lethargy, chronic fatigue, bruising,
osteoporosis, depression, hair loss, dental defects, short stature, infertility,
late puberty, early menopause, unsteady gait, muscle weakness [3-5].
At present, the only efficient treatment of CD in both symptomatic
and asymptomatic patients is a strict gluten-free diet throughout their
lifetime (plain meats, vegetables, fruits). Gluten is present not only in
cereals (wheat, rye and barley) and the baking products, but also in some
food ingredients including thickeners, stabilizers, flavourings, preservatives,
condiments, yeast extracts, etc. Unfortunately, gluten is also found in
inactive ingredients of pharmaceutical products [6]. Complete avoidance of
foods and medicines potentially contaminated with gluten is not an easy task
and needs a multidisciplinary approach from patients, doctors, dieticians and
pharmacists.
Despite that few pharmaceuticals are labelled as gluten-free, the
great majority of drug manufacturers do not specify on the product label
information regarding the gluten content [7]. A great concern comes from
the generic drugs, which may not contain the same excipients as the brand
product.
Detection of gluten in products is based on modern bioanalytical
methods e.g. enzyme-linked immunosorbent assay (ELISA) and
immunochromatographic techniques [8-9]. In the present paper, we
investigated the content of potentially hidden gluten ingredient in basic
pediatric drugs using the immunochromatographic visual test. This assay is
a qualitative and semiquantitative sensitive method for the detection of
gluten acting in good correlation with the ELISA assay [10].
Materials and methods
Sample extraction and preparation
Ten pharmaceutical products (pediatric syrups) purchased from the
national pharmacy market were used in the present investigation, as follows:
analgesics and anti-inflammatories (Panadol®, Paracetamol®), digestive
system drugs (Debridat®, Metoclopramid®, Duphalac®), antihistaminic
(Peritol®, Aerius®) and respiratory system drugs (Eurespal®, Rofedex®,
Fluidol®).
The samples were extracted in ethanol 60%. After 30 minutes
incubation, the samples were filtered and diluted 1:10 with phosphate-buffer
saline (PBS) containing 0.09% sodium azide, pH 7.4.
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Gluten detection
The immunochromatographic visual assay was used for qualitative
determination of gluten in pharmaceuticals. Immunochromatographic sticks
(purchased from OPERON S.A., Spain) which contain monoclonal
antibodies specific for gluten of wheat, barley and rye, were dipped into
solution samples diluted 1:10. Results were read after 5 min. of
chromatographic running. Development of a red band in addition to the
control blue band on the reaction strips is considered a positive result.
Results and discussion
Celiac patients can manage their condition only by a strict
gluten-free diet. Because of less obvious sources of gluten and lack of
specification of wheat in the ingredient listing, celiac patients must check
carefully the ingredients of food products and pharmaceuticals. It is known
that small amounts of gluten cause intestinal damages in these individuals.
Despite few common pediatric gluten-free drugs that are known, certain
generic drugs may not be gluten-free.
In the study, we have screened the presence of gluten in common
pediatric medications (analgesics and anti-inflammatories, digestive system
medications, antihistaminic, respiratory system medications) with unknown
gluten status. These samples are not recognized as devoid of gluten by the
celiac disease patients.
Samples were extracted with ethanol 60% and diluted 1:10 in PBS.
Qualitative determination of gluten was performed using the sensitive
immunochromatographic test. The gluten reaction strips consist of four
zones: the inferior adsorbent zone that is submerged in the sample, the red
microsphere zone, the reaction zone and the superior adsorbent zone. The
gluten positive samples react first with the red particles from the second
zone of the strip and then migrate in the reaction zone of the strip in order to
react with the anti-gliadin specific antibodies, developing a red band. A
control band (blue) appears in both positive and negative samples. The
sensitivity of the test is 10 ng/ml of gliadins equivalent to 2 ppm of gluten in
the sample.
The obtained results are given in table I.
As it isshown in table I, we obtained a negative gluten test for all
the samples. Consequently, the tested products do not contain gluten in
concentration above 2 ppm, the sensitive limit of the test. These
informations are important for completion of the existing lists of gluten-free
medications
(Wheaton
Gluten
Free
Support
Group).
The
immunochromatographic test used in the present gluten detection proved to
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FARMACIA, 2009, Vol.LVII, 1
be a sensitive, simple and reliable test for rapid determination of gluten in
pharmaceutical products.
Table I
Gluten content of selected pediatric pharmaceutical products
determined by immunocromatographic assay
Product name
Active ingredient
Form
Manufacturer
Gluten
Panadol baby®
Acetaminophen
susp.
Glaxo Smith
Klein
Paracetamol®
Acetaminophen
oral sol.
Terapia
Debridat®
Trimebutine
granules for
Pfizer
oral susp.
Metoclopramid
Metoclopramide
oral sol.
Pharmaplant
®
Biogalenica
Duphalac®
Lactulose
oral liq.
Solvay Pharma
Peritol®
Ciproheptadine
syrup
Egis
Pharmaceuticals
LTD.
Aerius®
Desloratidine
syrup
Schering Plough
Eurespal®
Fenspiride
syrup
Servier
hydrochloride
Rofedex®
Dextromethorphan
syrup
Biofarm
Fluidol®
Carbocysteine
oral sol.
TIS Farmaceutic
Note: "-" means negative gluten test
Conclusions
Celiac disease (CD) is a chronic intestinal disorder of public health
concern as untreated celiac patients develop disease complications and
suffer from the negative impact on their quality of life. The disease results
from ingestion of gluten from wheat, barley and rye. Gluten can be found
not only in food products, but also as normal component of drugs.
Detection of gluten in pharmaceuticals is a very important task
required for displaying the gluten status of products, as the absence of
gluten remains the only treatment for celiac patients.
In this paper, we investigated the presence of gluten in ten common
pediatric
medications
using
a
sensitive
procedure,
the
immunochromatographic assay. The results provide information about the
absence of gluten in the selected products contributing to the completion of
the existing lists of gluten-free pharmaceuticals. It is known that, for celiac
patients, obtaining accurate information about the gluten content of a
particular pharmaceutical is a difficult and time-consuming process.
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1.
Manuscript received: 18.06.2008