J. Vet. Med. A.

42, 177-183 (1995)

@ 1995 Blackwell Wissenschafts- Verlag, Berlin
ISSN 0931-184X

Catedra de Farmacologia, Facultad de Veterinaria, Uniwersidad Complutense de Madrid,

Spain

Comparison of the Kinetics of Sodium Meclofenamate

Versus Meclofenamic Acid after Oral Administration to
Sheep
T. ENCINAS,E. VINAGRE,J. C. BOGGIO, M. L. DE VICENTE,M. I. SAN ANDRESand
C. RODRIGUEZ

Address of authors: Dra. Teresa Encinas Cerezo, Citedra de Farmacologia, Facultad de

Veterinaria, Universidad Complutense de Madrid, Avda. Puerta de Hierro s/n 28040, Madrid,
EspGa

With 2 figures and 1 table

(Received for publication July 8, 1994)

Summary
Meclofenamates are non-steroidal anti-inflammatory agents used in ruminants for the
prevention and the treatment of anaphylactic processes. The objective of the present work was
to study possible kinetic variations due to the chemical form of meclofenamates administered
by the oral route to adult sheep. Six Rubia del Molar female sheep (2-3 years old, 47-57 kg)
were used. Initially, an intravenous administration of sodium meclofenamate (2.2 mg/kg bwt)
was given; the obtained kinetic results were in agreement with data from other authors. Oral
administrations (20 mg/kg bwt) of sodium meclofenamate and meclofenamic acid were then
given. When the reticular groove was opened, both drug forms showed a single meclofenamate
plasma peak; tZmuwere 60.0 -C 10.61 min and 127.50 -C 22.5 min for the sodium and acid form,
respectively. The elimination rate constants (B) were not significantly different, but the absorption
half-lives were (14.69 -C 3.21 min for the sodium form and 6 1 . 0 7 c 21.7 min for the acid
form). The bioavailability was 48.6 k 4.3% for sodium meclofenamate and 65.1 f 2.8% for
meclofenamic acid. Thus, the chemical form (sodium versus acid) alters the oral bioavailability
and t, of meclofenamates in adult sheep. These findings agree with the behaviour of meclo-
fenamates in man.

Introduction
Meclofenamic acid [N-(2,6dichloro-m-tolyl) anthranilic acid] and its sodium salt
(sodium meclofenamate) are non-steroidal anti-inflammatory agents. These com-
pounds have anti-inflammatory and antipyretic actions in horses and ruminants and
may have clinical value in the treatment and prophylaxis of allergic diseases, mastitis
and parturition induction in ruminants (WELLSet al., 1973; BURKAand SCARNELL,
1978; MITCHELL and FLINT,1978; MITCHELL et al., 1990; LOHUISet al., 1991). It has
been demonstrated that the efficacy of meclofenamates was directly related to the
plasma levels of the free drug (AITKENet al., 1975). I n human beings, the acid form

U.S.Copyright Clearance Center Code Statement: 0931 - 184X/95/4203 - 0177$11.00/0

178 ENCINASet al.

is more slowly absorbed than the sodium salt but the blood levels achieved are sustained
for a longer time (FLOREZ, 1992).
In cattle, studies with meclofenamates after intravenous, oral, intraruminal
(AITKEN and SANFORD, 1975) and intramuscular administration (MARRINERand
BOGAN, 1979) were reported. In sheep, however, there are only published works with
sodium meclofenamate administered by the oral and intramuscular routes (MARRINER
and BOGAN, 1979). In their study, a biphasic absorptive pattern was described, which
could be due to some of the drug by-passing the rumen, through the reticular groove.
The closure of the groove may be due to the stimulation of the oropharyngeal reflex
by the sodium salt (COOKEand NICHOLSON,1981).
The purpose of this work was to study the kinetic of the meclofenamates admin-
istered by the oral route to sheep, the effect of the chemical form (sodium versus acid)
and the influence of the reticular groove closure, using the glucose-test method
(ENCINAS, 1993).

Materials and Methods

Animals
Six Rubia del Molar female sheep aged 2-3 years and weighing 47-57 kg were used. Sheep
were kept indoors and deparasitised 2 months before trials. Room temperature, humidity, and
diet were maintained at a constant level.
A crossover design with a 2 week interval between trials was used for the experiments, each
sheep being used in each of the treatments in the study.
Drug and sample collection
Sodium meclofenamate (Parke & Davis) was dissolved in water :propylene glycol 250 (75 :25)
and meclofenamic acid (ARQUEL, Parke & Davis) in tepid water. Sodium meclofenamate was
administered intravenously (jugular) to the sheep at a dosage of 2.2 mg /kg bwt (active ingredient).
Jugular venous blood samples were taken from the conualateral vein with a syringe. Blood
samples were collected at 5, 10, 20, 30, 45, 60, 120, 240, 360 and 480 min and 24 h after
administration.
Meclofenamic acid (20 mg/kg bwt; active ingredient) and glucose (0.625 g/kg) were co-
administered orally with a drenching-gun. Blood samples were taken for drug determination at
5, 10, 15, 30, 45, 60, 90, 120, 150, 180, 300, 420, 540 and 660min and 24 and 48 h. Sodium
meclofenarnate (20 mg/kg bwt; active ingredient) and glucose (0.625 g/kg bwt) were co-admin-
istered orally with a drenching-gun. Blood samples were taken 5, 10, 15, 30, 45, 60, 90,
120, 150, 180, 300, 420, 540 and 660 min and 24 and 48 hours after sodium meclofenamate
administration.
The blood samples were introduced into heparinized tubes. Some of the sample (32 $)
which were collected after drug administration were used to determine glucose concentration
and plasma samples were collected after centrifugation (2OOOg, 20 min) and frozen.
Meclofenamate determination
A test tube, containing 0.5 ml of plasma, was mixed with 0.5 ml HCI (6N) and agitated for
2 min and then 5 ml methylene chloride was added. The tube was agitated again (5 min) and
centrifuged at 20OOg for 10 min. The organic layer (4 ml) was transferred to another test tube,
evaporated to dryness (40 "C, nitrogen stream), re-extracted in 200 yl mobile phase and a 20 $
volume was injected into the HPLC system (ENCINAS,1993).
Meclofenamate concentrations in plasma samples were determined by high-performance
liquid chromatography (HPLC: pump, KONTRON 420 and twinwave detector, WATERS
484) following the analytical conditions reported by ENCINAS(1993). A column (25 x 3.9;
10 pm) with C18 material (Spherigraph) and a precolumn (15 x 3.2; 10 pm; Brownlee) were
used. The mobile phase was acetonitrile:phosphate buffer (pH = 2.5) (60:40); the flow rate,
1.5 ml/min and the injection volume 20 pl. Detection was by UV absorption at 226 nm. Under
these conditions, retention time for meclofenamate was 5.10 min, recovery 90.26'70, intra-assay
variation 2.59%, inter assay variation 7.35% and the limit of detection was 0.17 pg/ml.

Reticular groove closure

During the oral trials, a glucose plasma concentration curve (glucose test) was performed
 Comparison of Sodium Meclofenamate Versus Meclofenamic Acid in Sheep 179

to assess the reticular groove state (closed or opened). A glucose peak of 5-15 min indicated that
the reticular groove was closed, whereas no peak for 45 min indicated that the groove was open,
according to the results reported by ENCINAS(1993).

Pharmacokinetic calculations
Concentration versus time curves for meclofenamate for each individual animal were fitted
with the software PKCALC, installed on a PC computer (SHUMAKER,1986). The following
equations were used to describe the concentration time curves for meclofenamate:
+
Cp = -0 e-v A e-M + B e-h
were: Cp = plasma concentration at time t after administration (pg/ml).
0, A, B = concentration at time zero extrapolated from the absorption and elimination
phases (pg/ml).
r,~, a, = exponents (per min) (BAGGOT,1977).
The choice of the best fittings was based on the determination of the coefficient values (2).
Non-compartmental analysis was used to calculate the area under the moment curve
(AUMC), area using trapezoidal rule (AUC), the mean residence time (MRT) and the bioavail-
ability (F). The mean residence time was determined as:
MRT = AUMC / AUC
The bioavailability was determined as:
F = De x AUCo / Do X AUCe
where: De = intravenous dose (mg/kg); Do = oral dose (mg/kg); AUCe = intravenous area
under the curve (pg min/ml); AUCo = oral area under the curve (pg min/ml).
Clmu,CZmrtlmuand tzmuwere reported as the observed values: Clmu = Peak concentration
when the peak appeared by 30 min; CZmu= Peak concentration when the peak appeared after
30 min; tlmu= Peak time when the peak appeared by 30 min; tzmu= Peak time when the peak
appeared after 30 min; Vd,, = Steady-state volume of distribution; C1= Clearance. (POWERS,
1990)

Statistical analysis
The pharmacokinetic parameters, determined for each individual animal, were reported as
the mean (k SEM) of six animals. The effects of the chemical form of the drug on meclofenamate
pharmacokinetics were calculated by non-parametric methods. The Wilcoxon signed ranks test
for paired observations was used to compare the sodium meclofenamate versus the meclofenamic
acid data. The results were considered significant when P =s 0.05.

Results
Mean plasma concentrations ( 2SEM) of meclofenamate as a function of time after
oral administration of sodium meclofenamate (20 mg/kg bwt) and meclofenamic acid
(20 mg/kg bwt) are illustrated in Figures 1 and 2, respectively. T h e pharmacokinetic
data are shown in Table 1.
Results of the glucose tests suggested that during the experiment with meclo-
fenamic acid not all animals closed the groove (no glucose peak), but during sodium
meclofenamate administration by the oral route, several animals (n = 2) closed the
groove (one glucose peak at 5-15 min). For that reason, results from the sodium
meclofenamate trial are presented from animals that closed the reticular groove (RGC)
and animals that did not close the groove ( R G O ) separately.
When sodium meclofenamate was administered and the reticular groove was
closed (RGC), two meclofenamate peaks were apparent: o n e at 15.0 2 0.0 rnin (Clmu=
24.01 f 0.63 pg/ml) and another one at 52.5 2 7.5 rnin (CZmu *
= 11.08 0.51 pg/ml).
When the groove remained opened ( R G O ) after the salt administration, a single plasma
peak was observed (Czmur= 6.92 & 1.47 pg/ml, tZmax = 60.00 2 10.61 min).
When meclofenamic acid was administered, a single plasma peak was observed
always at tzmw= 127.50 +- 22.5 min and Czmu= 6.49 0.54 pg/ml. *
180 ENCINASet al.
30 ~

I
0 120 240 360 400 600
I
Time (mln) IhJ

Fig. 1. Mean plasma concentrations following oral administration of sodium meclofenamate

(20 mg/kg bwt) to six sheep: (W) reticular groove opened (n = 4) and (+)reticular groove closed
(n = 2). (Mean 2 SEM).

7 --

6-

z53 5-

0 4 M k l k
400 600
+ 1%
0 120 240 360
Time (min) lh)

Fig. 2. Mean plasma concentrations following oral administration of (A)meclofenamic acid

(20 mg/kg bwt) to five sheep (reticular groove open) (Mean 2 SEM).

The single peak time (tzmlx)was significantly greater (P s 0.05) for meclofenamic
acid (127.5 5 22.5 min) than for sodium meclofenamate (60.00 t 10.61 min) after
drug administration, and the CZmudid not differ (P 2 0.05) between trials (6.49 t
0.54 pg/ml and 6.92 t 1.47 pg/ml, respectively). The bioavailability was significantly
greater for the acid form (65.10 t 2.77%) than for the sodium salt (55.39 t 10.23%;
Table 1).
Table 1. Mean pharmacokinetic parameters for sodium meclofenamate (2.2 mg/kg bwt) after single intravenous administration (n = 6) ('ENCINAS,
1993) and for meclofenamic acid (n = 6) and sodium meclofenamate (reticular groove opened (RGO; n = 4) and reticular roove closed (RGC;
n = 2)) after oral administration (20 mg/kg bwt). (Mean f SEM). Where: 0,A, B-concentration at time zero extrapolate!i from the absorption
and elimination phases; q, a, /%exponents; AUC-area under the curve; q,, tfor qghalf-lives of absorption and elimination; AUC-
area under concentration-time curves; MRT-mean retention time; VD,,-steady-state volume of distribution; Cl-clearance; C,,,-peak
concentration when the eak appeared by 30 min; Czm,-peak concentration when the peak appeared after 30 min; tlmu-peak time when the
p e g appeared by 30 min; tzm,-peak time when the peak appeared after 30 min; F-bioavailability
Sodium Meclofenamate Sodium Meclofenamate Meclofenamate
*Intravenous Oral RGO (20 mg/kg) Oral RGC (20 mg/kg) Oral RGC (20 mg/kg)
PARAMETER 2.2 mg/kg (n = 6) (n = 4) (n = 2) (n = 6)
- 19.29 5 8.66 - 8.82 f 1.26
30.29 f 3.40 10.84 4.91
* 21.58 1.54
* 4.36 f 1.26
1.62 f 0.05 1.63 0.19
* 4.88 f 0.22 4.44 2 1.02
- 5.42 1.00
* - 1.80 0.65
*
10.00 f 0.80 7.74 0.30
* 2.52 f 0.25 4.22 0.80
* 2
1.32 f 0.08 6.04 f 1.00 9.46 1.60
* 8.55 1.22
* ul
- 14.69 5 3.21 - 61.07 2 21.7 E
7.18 f 0.63 123.40 f 35.08 27.75 * 2.79 198.08 59.21
*
542.35 f 35.73 1336.15 358.09
* 755.30 2 129.24 861.08k 121.76
868.00 135.19
* 3078.26 t 276.72 6058.12 5 1291.87 5136.86f 218.50
617.92 49.09
* 1519.28 544.32
* - 1160.53 f 97.44
1679.95 5 398.96 7656.67 f 1829.20 - 3578.19 f 495.19
2.47 f 0.39 2.24 2 0.37 1.72 2 0.10 3.21 ? 0.98
- 60.00 f 10.61 52.50 k 7.50 127.50 f 22.5 ?
h
6.92 f 1.47 11.08 5 0.51 6.49 0.54
* 5'
- 15.00 0.00
* - v1
- 24.01 0.63
* - R
48.62 * 4.34 55.39 10.23
* 65.10 * 2.77 43
182 ENCINASet al.

Discussion
In previous works, it has been shown that the reticular groove closure could
modify the kinetic disposition of drugs including meclofenamates (MARRINER and
BOGAN, 1979; ENCINAS,1993). Therefore, whenever drugs are administered by the
oral route to ruminants, the effect of reticular groove closure must be considered
(SUITER et al., 1993).
In the present study, the reticular groove only closed in some animals (n = 2)
during oral administration of sodium meclofenamate, but it did not close during
meclofenamic acid administration. Closure could have been due to the oropharyngeal
reflex, provoked by the sodium salt of the drug (AITKENand SANFORD,1975). During
pharmacokinetic trials, whenever drugs are administered by the oral route, the status
of the reticular groove should be controlled (COOKEand NICHOLSON, 1981).
The tZrnax (60.0 min) after the salt administration was longer than described in
cattle (30 min) (AITKENand SANFORD, 1975) and sheep (40 min) (MARRINER and
BOGAN,1979) after oral administration of sodium meclofenamate (20 mg/kg). Greater
values, with a tmaxof 8-12 h were reported following intraruminal administration of
sodium meclofenamate (10 mg/kg) to adult cattle (AITKENand SANFORD,1975).
The CZmax (6.92 pg/ml) after sodium meclofenamate administration was greater
than described in adult cattle (3 pg/ml) and lower than in unweaned calves (9-10 pg/ml)
(MARRINER and BOGAN,1979).
The tzmX after sodium meclofenamate administration (60.0 min) was significatively
shorter than after meclofenamic acid trial (127.00 min), although did not show
any difference (P 3 0.05). Similar observations attributed to the different pK, were
reported for these two chemical forms administered to humans (FLOREZ,1992) and
these were attributed to the different pK, values of meclofenamic acid and sodium
meclofenamate. Previously, it has been suggested that non-ionized lipid-soluble drugs
only can be absorbed by passive diffusion through the wall of the gastrointestinal tract
(DOBSON, 1967).
Although not examined in the present study, the pH of ruminal fluid was reported
to be 5.5-6.5 and that of abomasal fluid 3-4. The pK, of meclofenamic acid is 4.0 and
that of sodium meclofenamate is 3.2. The equilibrium between plasma :rumen fluid
may be 15.7 :1 for meclofenamic acid and 15.8 :1 for sodium meclofenamate and the
equilibrium between plasma :abomasal fluids may be 1200 :1 and 3350 : I, respectively.
It seems therefore that drug absorption from the rumen occurs slower than the transfer
across the abomasal epithelium (DOBSON, 1967). The plasma peaks registered in the
present study are probably due to abomasal absorption of a part of the drug solution
that went to the abomasum through the top rumen liquid layer, since it has been
determined that the time to achieve this compartment is approximately 1 h (FAICHNEY,
1984).
The bioavailabilities of the two meclofenamates in both experiments were lower
than in humans (>%)YO) (FLOREZ,1992). Meclofenamic acid administered by the oral
route to sheep showed a bioavailability of 65.10%, similar to the bioavailability in
horses (SULLIVAN and SNOW, 1982) and greater than that of sodium meclofenamate in
sheep (48.62%). However, the peak time for the acid was later than for the salt and
plasma concentrations after sodium meclofenamate administration was maintained
over 2 pg/ml (minimum therapeutic concentration; AITKENet al., 1975; BURKAand
SCARNELL, 1978) during a longer period than after acid administration. Therefore, the
oral bioavailability of the meclofenamates in sheep depends on the chemical form of
the drug.

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