Acta Poloniae Pharmaceutica Drug Research, Vol. 64 No. 6 pp.

565567, 2007

ISSN 0001-6837
Polish Pharmaceutical Society

URIC ACID PLASMA LEVEL AND URINE pH IN RATS TREATED

WITH AMBROXOL
TOMASZ DREWA1,2, ZBIGNIEW WOLSKI2, MARZENA GRUSZKA3, BARTOSZ MISTEREK2
and JOANNA LYSIK4
Department of Tissue Engineering, Chair of Medical Biology,
Department of Urology, 3 Hospital Laboratory, University Hospital, N. Copernicus University,
Bydgoszcz, Poland,
4
Department of Oncology and Brachytherapy, Oncology Center, Bydgoszcz, Poland
1

Abstract: It was a chance discovery that ambroxol parenteral administration led to urinary bladder stone formation in rats. This study was undertaken to examine the serum uric acid levels and urine pH in rats after
ambroxol parenteral treatment. Ambroxol influence on the uric acid level was measured in 5 rats (Rattus sp.)
treated with 60 mg/kg (dissolved in injection water, sc, daily) during 2 weeks. Ambroxol influence on urine pH
was examined on 45 rats divided into 3 groups. Rats from the 1st and 2nd group received 30 and 60 mg/kg/24h
ambroxol, respectively. Urine was collected once daily and measured with strip kit. All values were presented
as the means with standard deviations. The Student t test was used to compare the means, p < 0.05 was considered as significant. Dynamics of pH changes was measured in 4 rats treated with 60 mg/kg/24h of ambroxol.
Controls received 1 mL of injection water sc. Serum uric acid level increased up to 8.7 1.0 mg/dL vs. 5.7
1.0 mg/dL in control (p < 0.002). In the 1st and 2nd group urine pH increased up to 7.5 0.5 and 7.6 0.5 vs.
6.7 0.4 (p < 0.05). Ambroxol withdrawal resulted in sequential urine pH decrease. 11 days after interruption
of ambroxol therapy pH reached the starting value. Urine pH changes and possible disturbances in uric acid
metabolic pathway may influence on the stone formation in rats after ambroxol parenteral treatment. The influence of ambroxol on urinary tract GAG layer and the balance between xanthine and CaOx in the urine should
be checked.
Keywords: drug-induced stones, ambroxol, hypoxanthine, calcium oxalate

mals had free access to chow (Murigran) and tap water.

Chow was obtained from Agropol and BASF (Poland).
Murigran was composed of protein (23%), raw fat
(3%), raw ash (7.5%), raw fibers (5%), lysine (1.5%),
methionine and cysteine (0.8%), tryptophan (0.15%),
calcium (1.1%), phosphorus ((0.7%), natrium (0.2%) ,
vitamins A, D, and E and cuprum. The 6 a.m. 6 p.m
day and night cycle was maintained artificially.
To establish the ambroxol influence on the
purine metabolism the uric acid level was measured. 5
rats were treated with commercially available ambroxol (Mucosolvan, Boehringer Ingelheim) subcutaneously (60 mg/kg sc, daily) during the period of two
weeks. Ambroxol was dissolved in injection water
always prior to injection. The dissolved drug was
injected under the skin. During the injection procedure
small enlargement tissue under the skin was observed.
Rats from the control group received 1 mL of injection
water sc. After two weeks animals were sacrificed and
2 mL of blood form each was obtained. 5 rats were
enrolled into the control group. Measurements were

Drug crystals are identified in two thirds of

drug-induced stones. Other stones have an apparent
metabolic origin (1). Ambroxol (ABX) is one of the
most often prescribed drug (2, 3). It was a chance
discovery that ambroxol parenteral administration
led to stone formation within a period of two months
in half of treated rats. Stones were composed of 67%
of xanthine and 33% of calcium oxalate (CaOx) (4,
5). Previous studies had suggested that high doses of
ambroxol reduced the plasma uric acid concentration by increasing its clearance (6).
Stone formation is a complex multifactor phenomenon. It is often difficult to find factors responsible for lithogenesis. This study was undertaken to
examine the serum uric acid levels and urine pH in
rats after ambroxol parenteral treatment.
EXPERIMENTAL
Wild rats (Rattus sp.) were used in the experiment.
1 month before and during the whole experiment ani-

* Correspondence: Tomasz Drewa, MD, PhD, FEBU, Department of Tissue Engineering, Chair of Medical Biology, N. Copernicus
University, Karowicza 24 str., 85-092 Bydgoszcz, Poland, fax: 0048 52 5853742, phone: 0048 52 585 3737; E-mail: tomaszdrewa@wp.pl

565

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TOMASZ DREWA et al.

performed, using commercially available UA plus kit

(Roche Diagnostics GmbH, Germany). The results
were presented as the means standard deviation. The
Student t test was used to compare the mean values, p
< 0.05 was consider as significant.
To examine the ambroxol influence on urine
pH 45 rats were divided into 3 equal groups. Rats
from the 1st and 2nd group received ambroxol during
2 weeks in daily doses of 30 mg/kg and 60 mg/kg sc
every morning, respectively. During the ambroxol
therapy rat urine was collected once daily. Urine pH
was measured with laboratory strip kit (SWW133128, POCh, Poland). All values were presented as the
means with standard deviations. The Student t test
was used to compare the mean values of pH in tested group. p < 0.05 was consider as significant.
To observe dynamics of pH changes after
ambroxol administration 4 rats were treated parenterally 10 days with a dose of 60 mg/kg/24h.
Then, the rats were observed during the next two
weeks, until the urine pH recovered to starting level.
Every day in the morning urine pH was measured
using strip kit (SWW1331-28, POCh, Poland).
RESULTS AND DISCUSSION
We have previously shown that ambroxol parenteral treatment induced lithogenesis in rats (5). In
humans, ambroxol is metabolized to dibromoanthranilic acid (DBAA) and 6,8-dibromo-3-(trans-4hydroxycyclohexyl)-1,2,3,4-tetrahydroquinazoline
(DHTQ). The ambroxol metabolism is similar in
human and rat (7). Previous studies had suggested
that high doses of ambroxol could reduce the plasma
uric acid concentration (6). In our study serum uric
acid concentration was elevated above the values of
control group (Fig. 1). Oosterhuis et al. had shown
that plasma hypoxanthine levels were not affected
by ambroxol (6). On the other side, it was observed
that some drugs administered in cardiovascular diseases elevated serum uric acid (8). In this study was

Figure 1. The serum uric acid level after 14 days of ambroxol parenteral treatment. Daily dose was 60 mg/kg/24 h. Animals in control group received 1 mL of phosphate buffered saline.

found that xanthine was one of the components of

urinary stones after ambroxol treatment. Xanthine
comprised about 67% of the stones weight.
Xanthine urinary stones are rare in humans. In the
control group serum uric acid level was 5.7 1.0
mg/dL. Serum uric acid level increased up to 8.7
1.0 mg/dL after two weeks of ambroxol parenteral
treatment in a dose of 60 mg/kg/24h. The difference
of serum uric acid level between control and treated
group was statistically significant, p < 0.002 (Fig.
1). The authors observed that xanthine urinary
stones in rats after ambroxol treatment had coexist
with elevated serum uric acid level, which is possible side effect of ambroxol treatment in rats.
Xanthine stones occurred in hypoxanthine-guanine
phosphoribosyltransferase deficiency, because of
accumulation of guanine (9). It was proved that partial deficiency of hypoxanthine-guanine phosphoribosyltransferase (HPRT) led to hypoxanthine and
serum uric acid accumulation (10, 11). It was shown
on in vitro model that uric acid metabolites accumulate in the case of HPRT deficiency (12). It can be
speculated that ambroxol treatment can affect the
uric acid metabolism. This pathway should be
examined in the future.
The urine pH is another important factor
responsible for lithogenesis. The H+ ions influence
the solubility of many substances in the urine. The
urine pH in the control group was 6.7 0.4. It was
noticed that average urine pH was higher than in
control after two weeks of ambroxol treatment. In
the 1st and 2nd group the urine pH increased up to 7.5
0.5 and 7.6 0.5, respectively. There were significant differences between experimental groups and
the control. No difference was observed between pH
values of two experimental groups (p = 0.28, Fig. 2).
Ten days of the continuous ambroxol treatment at
higher dose (60 mg/kg/24h) resulted in the pH
increase up to 7.6. Ambroxol withdrawal resulted in
sequential urine pH decrease. 11 days after interruption of the therapy pH reached the previous value

Figure 2. The rat urine pH after two weeks of ambroxol parenteral treatment.

Uric acid plasma level and urine dH in rats treated with ambroxol

567

Figure 3. Rat urine pH changes after 10 days of ambroxol parenteral treatment. Daily dose was 60 mg/kg/24h. Arrow indicates ambroxol
management interruption.

(Fig 3). It was observed in this study that pH

increased in a short period of time after ambroxol
administration. The next two weeks after ambroxol
withdrawal pH decreased to original value (Fig. 3).
It should be emphasized that both administrated
doses increased pH within similar ranges (Fig. 2).
Xanthine is a purine base. The solubity of xanthine
decreases concomitantly with an increase of pH.
The relationship between pH changes and purine
bases solubility is strong. The pH also effects the
solubility of calcium oxalate. Uric acid (UA) and
sodium urate (NaU) crystals could induce the precipitation of calcium oxalate (CaOx) from its inorganic metastable solutions (13, 14). It was shown
that cystine adding to undiluted human urine resulted in the marked enhancement of calcium oxalate
precipitation (15). When the concentration of the
components increases beyond the saturation level, a
state of super saturation exists in the urine, which is
thermodynamically unstable. Xanthine nucleation
may provoke precipitation of the calcium oxalate
from its solution (16).
It can be resumed that ambroxol parenteral
treatment leads to formation of stone comprised of
xanthine and calcium oxalate in a very short period.
The urine pH changes and possible disturbances in
uric acid metabolic pathway may influence on the
stone formation in rats after ambroxol parenteral
treatment. Thus, the influence of ambroxol on the
balance between xanthine and CaOx in the rat urine
should be checked.
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Received: 29.05.2007

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Erratum
In the paper of. S. Ray, K. Roy and Ch. Sengupta Acta Pol. Pharm. 64, issue 4 pp. 335-344 in the
title and in the text instead of Spirulina plantesis should be Spirulina platensis.