Plasma concentrations and behavioral,

antinociceptive, and physiologic effects

of methadone after intravenous
and oral transmucosal administration in cats
Tatiana H. Ferreira, MV, MSc; Marlis L. Rezende, DVM, PhD; Khursheed R. Mama, DVM;
Susan F. Hudachek, PhD; Antonio J. A. Aguiar, MV, PhD

Objective—To determine plasma concentrations and behavioral, antinociceptive, and phys-

iologic effects of methadone administered via IV and oral transmucosal (OTM) routes in
cats.
Animals—8 healthy adult cats.
Procedures—Methadone was administered via IV (0.3 mg/kg) and OTM (0.6 mg/kg) routes
to each cat in a balanced crossover design. On the days of drug administration, jugular
catheters were placed in all cats under anesthesia; a cephalic catheter was also placed
in cats that received methadone IV. Baseline measurements were obtained ≥ 90 minutes
after extubation, and methadone was administered via the predetermined route. Heart and
respiratory rates were measured; sedation, behavior, and antinociception were evaluated,
and blood samples were collected for methadone concentration analysis at predetermined
intervals for 24 hours after methadone administration. Data were summarized and evalu-
ated statistically.
Results—Plasma concentrations of methadone were detected rapidly after administration
via either route. Peak concentration was detected 2 hours after OTM administration and
10 minutes after IV administration. Mean ± SD peak concentration was lower after OTM
administration (81.2 ± 14.5 ng/mL) than after IV administration (112.9 ± 28.5 ng/mL). Seda-
tion was greater and lasted longer after OTM administration. Antinociceptive effects were
detected 10 minutes after administration in both groups; these persisted ≥ 2 hours after IV
administration and ≥ 4 hours after OTM administration.
Conclusions and Clinical Relevance—Despite lower mean peak plasma concentrations,
duration of antinociceptive effects of methadone was longer after OTM administration than
after IV administration. Methadone administered via either route may be useful for periop-
erative pain management in cats. (Am J Vet Res 2011;72:764–771)

P ain in cats has been inadequately managed because

of a lack of licensed drugs for treatment of pain in
this species and concerns related to adverse effects,
DIVAS
Abbreviations
Dynamic interactive visual analog scale
HR Heart rate
particularly the deficiency of certain hepatic uridine di- LOQ Limit of quantification
phosphate–glucuronyltransferase isoforms, which pre- OTM Oral transmucosal
disposes cats to adverse effects of some drugs.1 The use SDS Simple descriptive scale
of opioids in cats has been limited because these treat- TP Total plasma protein

Received February 7, 2010.
Accepted March 29, 2010.
From the Department of Anesthesiology, Botucatu Medical School
(Ferreira), and Department of Veterinary Surgery and Anesthesiolo-
gy, School of Veterinary Medicine and Animal Science (Aguiar), São
Paulo State University-UNESP, Botucatu, SP, Brazil, 18618-970; and
the Department of Clinical Sciences, College of Veterinary Medicine
and Biomedical Sciences, Colorado State University, Fort Collins,
CO 80523 (Rezende, Mama, Hudachek).
Supported by Fundação de Amparo à Pesquisa do Estado de São Paulo
(FAPESP) grant No. 07/59505-3.
The authors thank James R. ZumBrunnen for statistical assistance
and Felicia Balzano, Sheryl Carter, and Anna Kendall for technical
assistance.
Address correspondence to Dr. Mama (kmama@colostate.edu).
ments have been associated with excitement,2,3 despite
reports3,4 that suggest opioid administration results in
signs of analgesia and euphoria (ie, purring, rolling,
rubbing, and kneading with forepaws) when used ap-
propriately in cats with signs of pain.
To date, the analgesic effects of several opioids, in-
cluding morphine, buprenorphine, hydromorphone,
oxymorphone, and butorphanol, have been evaluated
in cats5–11; however, the use of methadone, a synthetic
opioid, in this species has received less attention. Early
reports12,13 indicated that this drug may potentially be
useful as an analgesic, likely because of its unique phar-
macological properties; methadone binds to µ opioid

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 peptide receptors and also has antagonist activity at N-
methyl-D-aspartate receptors. Antagonism of N-methyl-
D-aspartate receptors is reported12 to mitigate spinal fa-
cilitation of pain. Additionally, unlike other µ opioid
peptide receptor agonist opioids, methadone inhibits
the reuptake of serotonin and norepinephrine12–14 and
promotes the blockade of nicotinic cholinergic recep-
tors.15 These are additional mechanisms thought to play
important adjunct roles in methadone-mediated anal-
gesia.15 In cats, administration of methadone IM or SC
was reported to result in a rapid onset of analgesia after
surgery but to vary markedly in duration of effect.16–18
Mild sedation was reported by some authors,16,18 where-
as others did not detect this effect.17
The OTM route has some advantages over other
routes of drug administration because it is simple, pain-
less, and generally well tolerated. Additionally, if drug
characteristics are favorable, there is a potential for
rapid absorption, and first-pass elimination by the liver
may be minimized.19 The high acid dissociation con-
stant of methadone and the alkaline environment of the
cat’s mouth collectively make it likely that methadone
administered OTM has a high bioavailability,19 suggest-
ing that methadone administered OTM may potentially
be useful for pain management in cats.
The purpose of the study reported here was to mea-
sure plasma drug concentrations and compare behav-
ioral, antinociceptive, and physiologic effects of meth-
adone after IV and OTM administration in conscious
cats.
Materials and Methods
Animals—Eight healthy adult (1- to 2-year-old)
neutered mixed-breed cats, including 4 females and 4
males, with a mean ± SEM body weight of 5.91 ± 0.45
kg, were used in this study. Cats were housed as a group,
with fresh water and commercial dry cat food available
ad libitum. Before the study began, cats were handled
for familiarization with study personnel, procedures
(including nociceptive devices), and environment,
which was located adjacent to their group housing. On
the days of drug administration, cats were individually
housed but interacted frequently with study personnel.
The study was approved by the Institutional Animal
Care and Use Committee of Colorado State University.
Experimental protocol—A balanced crossover de-
sign was used in which methadonea was administered
IV (0.3 mg/kg) and OTM (0.6 mg/kg) to each cat, with
a minimum of 10 days in between the 2 treatments.
Thus, 4 of 8 cats were assigned to receive either treat-
ment first and were assigned to the IV or OTM group
on each treatment day according to the method of drug
administration. Before methadone was administered
OTM, oral cavity pH was measured by use of pH paperb
placed in contact with the buccal mucosa.
On the days of drug administration, cats were anes-
thetized by use of a chamber, mask, or both with 5%
isofluranec in oxygen (5 L/min) delivered via a standard
small animal circle breathing system until orotrache-
al intubation could be performed with a cuffed tube.
During anesthesia, HR and rhythm were monitored
by use of an ECG, and systolic arterial blood pressure
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was monitored by use of an ultrasonic Doppler flow
detector.d
A 16-gauge cathetere was aseptically placed in a
jugular vein by use of an over-the-wire technique and
secured with 2-0 monofilament nylon-polyamide su-
ture and a bandage to facilitate subsequent blood sam-
ple collection for analysis of plasma drug concentra-
tions. Cats in the IV group also had a 22-gauge catheterf
aseptically inserted into a cephalic vein for administra-
tion of methadone. The cephalic catheter was removed
approximately 1 hour after drug administration. Iso-
flurane administration was discontinued after catheter
placement, and cats were continuously monitored dur-
ing recovery. Total anesthesia time (from administra-
tion of isoflurane until extubation) was recorded.
A minimum of 90 minutes after extubation, cath-
eter patency was verified, baseline blood samples were
collected, and methadone was administered. For OTM
administration, 0.6 mg of methadone/kg was delivered
between the lateral gingival surface and buccal mucosa
with a 1-mL syringe. For IV administration, 0.3 mg of
methadone/kg was delivered as a bolus via the cephalic
catheter followed by a 2-mL heparinized saline flush.
The volume of blood collected during each experiment
was adjusted for each individual cat so that < 10% of
its total blood volume (67 mL/kg)20 was removed over
the duration of each experiment. On the basis of body
weight of cats in the study, this was between 2 and 2.5
mL/sample. A similar volume (2 to 2.5 mL) of heparin-
ized saline (0.9% NaCl) solution (4 U of heparin/mL)
was administered after each sample collection to flush
and maintain patency of the catheter. Blood samples
were obtained immediately prior to drug administra-
tion (ie, baseline) and at 2, 5, 10, 20, and 30 minutes
and 1, 2, 4, 6, 12, and 24 hours after drug administration.
Blood was transferred to tubes containing lithium heparin
that were refrigerated and centrifuged (1,016 X g) for 10
minutes at 4°C. Plasma was separated and stored at –70°C
until analysis. Packed cell volume and TP were evaluated
in samples obtained prior to and 1, 6, and 24 hours after
drug administration. For determination of PCV, a micro-
capillary centrifugeg was used. Total plasma protein was
measured by use of a refractometer.h
Heart rate, respiratory rate, sedation, behavior, and
response to nociceptive stimulus were assessed by a
single observer (THF) at baseline and at 10 and 30 min-
utes and 1, 2, 4, 6, 8, 12, and 24 hours after methadone
administration. Heart rate and respiratory rate were de-
termined by means of auscultation and observation of
thoracic excursions, respectively, each for a minimum
of 15 seconds. Rectal temperaturei was measured just
prior to and at 1 and 6 hours after methadone admin-
istration. Temperature was the last variable recorded at
each time point because of the potential for thermom-
eter insertion to influence the cat’s behavior.
To assess sedation, an SDS and a DIVAS were used.
To implement use of these scales, cats were observed,
then approached, spoken to, and petted. The SDS was
used to evaluate sedation on a scale of 0 to 4 as follows:
0, euphoric behavior (meowing, purring, rolling, rub-
bing, kneading with forepaws); 1, rising and moving
to the enclosure door to investigate activity, purring,
meowing; 2, resting in sternal recumbency but appar-
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 ently listening and responding with head movement
without rising; 3, apparently sleeping but responding
by opening the eyes and raising the head; and 4, sleep-
ing with no response to a single handclap. The DIVAS
consisted of a horizontal 100-mm line on which 0 mm
corresponded to normal behavior and consciousness
and 100 mm corresponded to unconsciousness. A
mark was placed on the line to indicate the degree of
sedation. Because use of the scales did not fully capture
additional behavioral responses in the same manner as
those described for euphoria, other notable behaviors
were also recorded.
Food and water were offered 1 hour after drug ad-
ministration. Feeding behavior, water consumption,
urination, and defecation were recorded. After drug ad-
ministration, changes in salivation, drooling, vomiting,
and other physical changes, such as mydriasis and third
eyelid protrusion, were also recorded.
Two mechanical nociceptive devices were used se-
quentially to determine the response to noxious stimu-
lus before and after methadone administration. The first
was a custom C clamp outfitted with a calibrated 1-cm2
force transducer connected to an electronic recorder
capable of recording the pressure (ie, force) at which
the cat first responded. The device was used to manu-
ally apply force in a dorsopalmar direction across either
metacarpus. The second, an algometerj with a 1-cm2
circular tip, was manually applied to the lateral surface
of either antebrachium, midway between the elbow
and carpus. The algometer was externally calibrated
and, similar to the C clamp, recorded the force at which
the first response was detected. Responses included the
cat turning its head toward the stimulus, moving away
from the stimulus, vocalizing, or attempting to bite.
The stimulations were immediately stopped when a re-
sponse was detected. To prevent tissue damage if cats
did not react, cutoff values were preset for the C clamp
(20 kg/cm2) and algometer (5 kg/cm2). These cutoff val-
ues were similar to or lower than those used previouslyk
in other species in our laboratory; in those studies, no
evidence of tissue damage or discomfort was detected.
All applications and evaluations were performed by
the 1 evaluator (THF). The recorded response-eliciting
force was accepted as the antinociception score for each
method at each time point.
Response to noxious stimulation was assessed
before induction of anesthesia with isoflurane and
immediately before methadone administration to
ascertain whether anesthesia had any effect on the
response. Because no significant difference was de-
tected between these responses the mean of these
values was considered the baseline for each cat. For
both statistical analysis and graphic representation,
postadministration values were normalized to those
obtained at baseline.
Analysis of plasma methadone concentrations—
The high-performance liquid chromatography systeml
consisted of a binary pump, vacuum degasser, column
compartment with thermostat, and autosampler.m The
high-performance liquid chromatography columnn
was protected by a cartridgeo and maintained at room
temperature (22°C). The mobile phase consisted of an
aqueous component of 0.1% formic acid in water and
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an organic component of acetonitrile. The 3.5-minute
run consisted of the following linear gradient elution:
75% formic acid and 25% acetonitrile at 0 minutes, 10%
formic acid and 90% acetonitrile at 3.0 minutes, 75%
formic acid and 25% acetonitrile at 3.1 minutes, and
75% formic acid and 25% acetonitrile at 3.5 minutes.
The system operated at a flow rate of 1.0 mL/min.
Mass spectrometryp was performed by use of mul-
tiple reaction monitoring. Ions were generated in posi-
tive ionization mode by use of an electrospray interface.
Methadone compound–dependent parameters were as
follows: declustering potential, 41.15 V; entrance po-
tential, 4.04 V; collision cell entrance potential, 12.03 V;
collision energy, 20.82 V; and collision cell exit po-
tential, 2.11 V. Fentanyl (internal standard) com-
pound–dependent parameters were as follows: declus-
tering potential, 38.64 V; entrance potential, 3.84 V;
collision cell entrance potential, 13.24 V; collision en-
ergy, 31.18 V; and collision cell exit potential, 2.39 V.
Source-dependent parameters were as follows: nebu-
lizer gas, 3.52 kg/cm2; auxiliary (turbo) gas, 4.22 kg/
cm2; turbo gas temperature, 550°C; curtain gas, 3.52
kg/cm2; collision-activated dissociation gas (nitrogen),
0.42 kg/cm2; ionspray voltage, 4,500 V; and interface
heater, 100°C. Peak area ratios obtained from multiple
reaction monitoring of methadone (mass-to-charge ra-
tio, 310.2 → 256.2) and fentanyl (mass-to-charge ratio,
337.1 → 188.1) were used for quantification. The LOQ
of the assay was 1 ng/mL.
Methadone and fentanyl standard solutions were
prepared in acetonitrile. Methadone was extracted from
plasma by adding 300 µL of acetonitrile to 100 µL of
sample plasma, vortexing for 10 minutes, and cen-
trifuging at 18,000 X g for 10 minutes. An aliquot of
10 µL of the supernatant was injected into the liquid
chromatography–tandem mass spectroscopy system for
analysis.
Statistical analysis—Data analysis was performed
by use of statistical software.q A randomized block design
with repeated measures was used. The blocking effect
was cat, the repeated measures effect was time, and treat-
ment was the between-cat effect. Comparisons between
antinociceptive responses before and after anesthesia
(used to establish the baseline for antinociceptive effects)
were analyzed by use of a randomized block design. For
repeated-measures ANOVA, post hoc pairwise compari-
sons between treatments at each time and between times
for each treatment were performed by use of Student t
tests. Because residuals for DIVAS, C clamp, and plasma
methadone concentration values were skewed, log (y + 1)
transformation was used. For algometer, C clamp, and
DIVAS data, analysis was performed only for the interme-
diate times because baseline and 4 of the 7-hour responses
were nearly all zeros. Because we used the LOQ for plasma
methadone concentration, baseline data for this variable
had no variation and were excluded from the analysis. Dif-
ferences were considered significant at a value of P < 0.05.
Values are expressed as mean ± SEM.
Results
Total anesthesia time (from first inhalation of iso-
flurane to extubation) for catheter placement was 59 ±
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 3.5 minutes and 57 ± 4.2 minutes for cats assigned to cats in the IV and OTM groups, respectively, had third
receive methadone via IV and OTM routes, respectively. eyelid protrusion, but this did not seem to be related to
Time from extubation until methadone administration their degree of sedation.
was 100 ± 5.3 minutes and 105 ± 3.2 minutes for cats Following a period of sedation, euphoric behavior
in the IV and OTM groups, respectively. Mean pH of the that ranged in duration from 2 to 6 hours after drug ad-
oral cavity before methadone administration was 8.8 ± ministration was observed in 6 of 8 cats in the IV group.
0.1. All cats urinated, defecated, and had apparently The remaining 2 cats had more affectionate behavior
normal appetites during the study period after metha- (eg, rubbing up against the handler, rolling over when
done administration via either route. None of the cats touched) than was known to be typical for them. In the
vomited during the study period. Except for 1 cat that OTM group, 7 of 8 cats had signs of euphoria for 6 to 12
began to salivate and tried to escape when methadone hours after methadone administration. However, prior
first came into contact with the oral mucosa, adminis- to the onset of euphoria, 2 cats in the OTM group were
tration via the OTM route was without incident. How- sensitive to noise, and 1 of these 2 had apprehensive be-
ever, an increase in salivation (drooling) was detected haviors (ie, appeared apprehensive and did not like to
from 1 to 60 minutes after OTM methadone adminis- be touched) immediately after drug administration. The
tration in 7 of 8 cats. In 3, 1, and 2 cats, the duration noise sensitivity lasted 30 minutes in 1 cat and 1 hour
was 2, 7, and 20 minutes, respectively. One cat drooled in the other, which also had the described apprehensive
intermittently for 60 minutes after methadone admin-
istration. Licking of nose and lips was observed in 5 of Table 1—Mean ± SEM plasma methadone concentrations (ng/
8 cats in the IV group and in 1 of 8 cats in the OTM mL) in 8 healthy cats before (ie, baseline) and at predetermined
group after administration of methadone. The duration time points after IV (0.3 mg/kg) or OTM (0.6 mg/kg) administra-
of this behavior ranged from 1 to 10 minutes after the tion of methadone.
drug was given. Administration group
Plasma concentrations of methadone in cats were
determined after the drug was administered via IV Time point (min) IV OTM
and OTM routes (Table 1; Figure 1). These values 0 (Baseline) 1 6 0 160
peaked at 10 minutes and 2 hours in the IV and OTM 2 63.7 6 16.8c,d 14.2 6 8.0a*
5 82.9 6 21.4 23.2 6 5.5b*
c,d,e,f
groups, respectively. Twenty-four hours after adminis- 10 112.9 6 28.5e,f 37.1 6 6.3c,d*
tration, methadone concentrations were significantly 20 89.6 6 9.0f 37.4 6 8.9c,d*
lower than those measured at 12 hours, regardless of 30 83.0 6 8.2e,f 58.6 6 19.6d,e,f*
the route of drug delivery; however, these values were 60 83.8 6 8.9e,f 77.9 6 18.5f,g
higher than baseline values. Plasma concentrations of 120 72.1 6 5.6d,e,f 81.2 6 14.5g
methadone were significantly lower at 2, 5, 10, 20, and 240 55.8 6 5.9c,d,e 77.1 6 13.0f,g
360 43.2 6 2.3b,c 63.9 6 10.3e,f,g
30 minutes after administration in the OTM group, 720 28.3 6 1.7b 43.4 6 6.9d,e
compared with values in the IV group. Changes in PCV 1,440 18.0 6 1.5a 25.4 6 3.9b,c
and TP over time were recorded (Table 2). No signifi-
cant difference was detected between the 2 groups. All cats received methadone via each route in a balanced
crossover design study with an interval of $ 10 days between the
Heart rate 30 minutes after methadone adminis- 2 treatments.
tration was significantly lower in cats of the IV group, *Indicates significant (P , 0.02) difference between groups.
compared with that in cats of the OTM group (Table 3).
a–g
Within a column, values with different superscript letters are
significantly (P , 0.05) different from each other. Baseline values
In cats of the IV group, HR was significantly decreased were considered the LOQ.
from baseline at 10 and 30 minutes after drug admin-
istration; no differences were found over
time in the OTM group. Although respira-
tory rates were similar between groups, a
significant decrease from baseline was ob-
served from 10 minutes through 6 hours
after drug administration in the IV group
and at 4 hours in the OTM group. No dif-
ferences were detected in body temperature
within or between groups.
The degree and duration of sedation
after methadone administration varied in
cats of both groups (Figure 2). After drug
administration, cats in the OTM group had
significantly higher DIVAS scores at 30
minutes and 1 hour and higher SDS scores Figure 1—Mean ± SEM plasma methadone concentrations determined in a bal-
at 30 minutes, compared with scores for anced crossover design study of 8 healthy cats before (ie, baseline) and at pre-
determined time points after IV (0.3 mg/kg; black squares) or OTM (0.6 mg/kg;
cats in the IV group. Onset of sedation was white triangles) administration of methadone. Each cat received methadone via
also variable and occurred between 1 and 5 each route with an interval of ≥ 10 days between the 2 treatments. Peak plasma
minutes in 6 of 8 cats in the IV group and drug concentrations were detected 10 minutes after IV administration of metha-
between 2 and 10 minutes in 7 of 8 cats done, whereas peak concentrations following OTM administration were detected
at 120 minutes. *Values were significantly (P < 0.05) different between groups (IV
in the OTM group. Three of 8 and 1 of 8 and OTM). B = Baseline.

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 Table 2—Mean ± SEM PCV, TP concentration, and rectal temperature evaluated in the 8 cats in Table
1 before (ie, baseline) and at predetermined time points after IV or OTM administration of methadone.

Time point (h)

Variable Administration group 0 (Baseline) 1 6 24
PCV (%) IV 35 6 1.2 32 6 1.3* 31 6 1.1* 32 6 1.0*
OTM 34 6 1.6 32 6 1.1 32 6 1.5 29 6 1.0*
TP (g/dL) IV 6.5 6 0.2 6.1 6 0.2* 6.2 6 0.2* 6.3 6 0.2*
OTM 6.3 6 0.2 6.1 6 0.2 6.3 6 0.2 6.2 6 0.2
Temperature (°C) IV 38.8 6 0.2 38.9 6 0.1 38.6 6 0.1 NA
OTM 38.4 6 0.2 38.5 6 0.4 38.5 6 0.2 NA

*Indicates significant (P , 0.04) difference within a group, compared with baseline value.
NA = Not applicable.

Table 3—Mean ± SEM values for selected physiologic variables
evaluated in the 8 cats in Table 1 before (ie, baseline) and at pre-
determined time points after IV or OTM administration of metha-
done.
Respiratory rate
HR (beats/min) (breaths/min)
Time point IV group OTM group IV group OTM group
Baseline 206 6 9 209 6 8 55 6 8 56 6 6
10 min 168 6 11† 188 6 12 40 6 10† 53 6 8
30 min 175 6 13† 204 6 12* 43 6 9† 45 6 9
1 h 184 6 14 201 6 16 42 6 6† 45 6 8
2 h 221 6 14 218 6 14 43 6 5† 45 6 7
4 h 201 6 11 223 6 14 44 6 5† 45 6 4†
6 h 200 6 9 213 6 11 42 6 5† 45 6 3
8 h 203 6 8 199 6 15 50 6 5 46 6 4
12 h 217 6 7 228 6 8 45 6 4 47 6 5
24 h 211 6 8 199 6 6 46 6 3 46 6 3
*Indicates significant (P , 0.02) difference between groups.
†Indicates significant (P , 0.04) difference within a group, com-
pared with baseline value.
behaviors for that same amount of time. Marked my-
driasis was observed in all cats in both groups within 1
to 2 minutes after methadone administration. Duration
of mydriasis was 8 hours in 5 of 8 cats in the IV group
and 3 of 8 cats in the OTM group and was 12 hours in
the remaining cats in each group.
The cats tolerated application of nociceptive de-
vices and had no signs of injury or change in activity
after the effects of methadone were no longer discern-
able. Significantly higher antinociception scores were
recorded via algometer at 10 minutes and 1 hour after
drug administration in the IV group, compared with
scores in the OTM group (Figure 3). For cats in the IV
group, these antinociception scores were significantly
increased from 10 minutes to 4 hours after drug ad-
ministration, and antinociception scores recorded via
C clamp were significantly increased from 10 minutes
to 2 hours, compared with baseline values. In the OTM
group, significant increases were detected in antinoci-
ception scores recorded via algometer from 10 minutes
to 6 hours and in antinociception scores re-
corded via C clamp at 10 and 30 minutes
and 4 hours. From 1 to 4 hours after metha-
done administration, 3 of 8 cats in the IV
group and 5 of 8 cats in the OTM group
had such marked euphoric behaviors that
applying the stimulus and interpreting the
response were challenging.

Discussion
The study reported here evaluated se-
lected behavioral, antinociceptive, and
physiologic effects of methadone adminis-
tered via IV and OTM routes in cats. Plasma
concentrations of methadone were evalu-
ated to begin to define the relationships
of methadone concentration and effect, al-
though pharmacokinetic properties of the
drug were not determined in this study.
Methadone doses used in the present
study were selected on the basis of reports
of other studies10,16,17; racemic methadone
has been administered to cats at doses rang-
Figure 2—Mean ± SEM sedation scores assessed by use of an SDS (A; range of pos- ing from 0.1 to 0.6 mg/kg. We selected a
sible scores, 0 [euphoric behavior] to 4 [sleeping and not responsive to a handclap]) dose of 0.3 mg/kg for IV administration and
and a DIVAS (B; range of possible scores, 0 [normal behavior and consciousness] to 0.6 mg/kg for OTM administration because
100 mm [unconscious]) before and after methadone administration in the 8 cats in Fig-
ure 1. Sedation scores were normalized to baseline values. †Values were significantly of the possibility that cats would swallow
(P < 0.05) different from baseline values within a group. See Figure 1 for remainder part of the dose, possibly rendering that
of key. portion unavailable for absorption or result-

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 SE plasma concentration of methadone
decreased to 3.78 ± 0.82 ng/mL, approach-
ing the LOQ of 2 ng/mL at 6 hours after
administration of a dose of 0.5 mg/kg, IV.
On the basis of these earlier studies, we
elected to discontinue sample collection
for plasma concentration analysis at 24
hours with the assumption that this would
include an adequate period for drug con-
centrations to return to baseline. However,
in cats of the present study, plasma metha-
done concentrations well above the LOQ
could be detected even at 24 hours after
administration via the IV and OTM routes
(18.0 ± 1.5 ng/mL and 25.4 ± 3.9 ng/mL,
respectively). Further investigation may
be beneficial in determining reasons for
this apparent difference in drug disposi-
tion among various species. Although we
removed < 10% of total blood volume in
each cat and cats were allowed free access
to food and water throughout most of the
study period, we detected significant de-
creases in PCV and TP from baseline val-
Figure 3—Mean ± SEM force response thresholds (antinociception scores) before ues; the values were still within normal
and after methadone administration in the 8 cats in Figure 1. Antinociception was
assessed via sequential application of force with 2 mechanical nociceptive devices limits for the species. These decreases
25

(a custom C clamp [A] applied at either metacarpus and an algometer [B] applied were likely attributable to collection of
at either antebrachium). Values were recorded for the amount of force that first blood samples and hemodilution caused
elicited a response from the cat. Baseline values for antinociception were taken
as the mean of scores assessed by application of the same stimulus prior to, and by administration of heparinized saline so-
after recovery from, induction and maintenance of anesthesia with isoflurane for IV lution used to flush the catheter.
catheter placement prior to methadone administration. Subsequent antinociception Heart rate was decreased from base-
scores were normalized to baseline values. See Figures 1 and 2 for key.
line by 18% and 15% at 10 and 30 minutes,
respectively, after methadone administra-
ing in extraction via first-pass metabolism in the liver. tion in cats of the IV group only. Although HR remained
Low values of systemic bioavailability have been associ- within or slightly above the reference interval for cats
ated with swallowing buprenorphine intended for OTM (168 to 221 beats/min)26 in this study, an 18% reduction
administration in humans. Because, to the authors’
21 may be relevant in a critically ill patient or one with a
knowledge, this was the first study to include OTM ad- lower starting value. The largest decrease in HR corre-
ministration of methadone in cats, we tried to ensure sponded with the highest plasma drug concentrations
that efficacy was not influenced by drug dose. The high in cats of the IV group. No changes in HR have been
acid dissociation constant and lipid solubility of metha- reported following methadone administered IM or SC
done make it well suited for absorption from the buccal (0.3 and 0.6 mg/kg, respectively) in cats.17,18 This infor-
mucosa in cats. However, in the present study, peak
19 mation is consistent with findings for the OTM group
plasma drug concentrations after OTM administration in the present study and likely a result of the gradual
were lower than those detected after IV administration, rise in plasma drug concentrations when methadone is
despite the fact that the dose given to cats in the OTM given via routes other than IV.
group was twice that given to cats in the IV group. Decreased respiratory rates were also detected in
Plasma drug concentrations also increased more slowly cats that received methadone IV. This is likely the result
after OTM administration than after IV administration, of changes from atypically high baseline values in these
consistent with slower absorption of the drug as is re- cats. After methadone administration, respiratory rate
ported in humans administered opioids via the sub- values remained within the normal range reported27 for
lingual route. Reabsorption from the gastrointestinal
19 cats in both the IV and OTM groups (40 to 50 breaths/
tract has also been suggested as a reason for later peaks min and 45 to 53 breaths/min, respectively). This is
in plasma concentrations of other drugs such as pro- consistent with the observations of other investigators
pranolol administered via the OTM route in humans, who reported16–18 the absence of changes in respiratory
compared with the time to peak plasma concentrations rates following methadone administration to cats.
following administration via other routes.22 Although sedation was not formally assessed un-
To the authors’ knowledge, no other study has re- til 10 minutes after drug administration, cats in both
ported plasma concentrations of methadone in cats. In groups appeared sedated very soon after drug admin-
horses, plasma drug concentrations were detected for istration. The DIVAS and SDS scores at 10 minutes
12 hours after administration of 0.1, 0.2, and 0.4 mg of supported this observation; analysis of DIVAS and SDS
methadone/kg, PO, when an LOQ of 2 ng/mL was used scores also indicated that sedation was of a signifi-
for analysis.23 In a study24 in Greyhounds, the mean ± cantly longer duration in cats of the OTM group (up

AJVR, Vol 72, No. 6, June 2011 769

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 to 1 hour) than in cats of the IV group (10 minutes).
Whereas sedation coincided with the peak in plasma
methadone concentrations in the IV group, onset of
sedation in the OTM group occurred prior to the mea-
sured peak in plasma drug concentration. This suggests
that despite lower plasma concentrations, methadone
distributed rapidly to the effect site and, as a result of
presumed association with receptors, resulted in mea-
surable responses. A similar rapid onset of action has
been reported with use of levo-methadone (0.3 mg/kg,
SC) in cats.18 Opioid administration can produce seda-
tion in dogs and cats28,29; however, this effect in cats is
usually nonexistent or mild, compared with the effect
in dogs.16–18,30,31 Therefore, it is interesting that in the
present study, marked sedation was observed in most
cats, albeit for a short duration.
Interestingly, while plasma drug concentrations
peaked between 2 and 4 hours after drug administra-
tion in the OTM group of the present study, SDS scores
for sedation decreased because of euphoric behavior.
Euphoria is commonly reported following opioid ad-
ministration in cats and is often accompanied by my-
driasis,2,3,6,10,11,17,18 which may be mediated by the re-
lease of catecholamines from the adrenal gland.32 In
the present study, marked mydriasis was observed in
cats in both groups for 8 to 12 hours after methadone
administration but did not appear to result in adverse
effects. Interestingly, the onset of mydriasis was de-
tected almost immediately after drug administration in
both groups and most often coincided in the early pe-
riod with sedation and later with signs of euphoria (eg,
purring, kneading, and rubbing against objects). Signs
of dysphoria (eg, excitement, anxiety, restlessness, hiss-
ing, clawing, and biting) were not observed following
administration of methadone via either route in the
study reported here. This is consistent with results from
other studies10,16,17 in which a similar dose of racemic
methadone was administered in cats.
As with sedation, antinociceptive onset seemed
to be related to plasma methadone concentrations in
cats of the IV group; however, the peak antinociceptive
effect was observed prior to detection of peak plasma
concentrations in the OTM group. The mechanical
nociceptive devices used in this study have not been
previously used in cats to our knowledge. However, re-
sults of the present study suggest that both mechanical
devices, when applied by 1 evaluator, induce consis-
tent and repeatable responses at baseline; methadone
administration resulted in an increased nociceptive
threshold, which then gradually returned to baseline
values. Duration and magnitude of analgesia did not
appear to correspond to plasma drug concentration.
For example, in cats of the OTM group, a significant
antinociceptive effect was seen at 10 minutes after
methadone administration when mean plasma concen-
tration of the drug was 37.1 ± 17.8 ng/mL, whereas at
12 hours, a methadone concentration of 43.4 ± 19.4 ng/
mL was not associated with significant antinociception.
Modeling pharmacokinetic and pharmacodynamic ac-
tivity of the drug would allow a better assessment of
whether plasma drug concentrations can be used to as-
sess antinociceptive effects in cats. In human patients,
the mean minimum effective analgesic plasma concen-
770 AJVR, Vol 72, No. 6, June 2011
10-02-0044r.indd 770
tration of methadone is 59.2 ng/mL.33 However, the
range of plasma methadone concentrations (22 to 89
ng/mL) reported prior to recommending administra-
tion of additional analgesics is broad.33
It has been suggested that genetic polymorphisms
in enzymes may account for variations in metabo-
lism, analgesic efficacy, and adverse effects of metha-
done among humans.13 Similar diversity in responses
to methadone administration was identified in cats of
the present study; mean antinociceptive effects lasted
2 and 4 hours in IV and OTM groups, respectively, but
some cats had detectable antinociceptive effects for up
to 8 hours, and others had these effects for only 1 to
2 hours. One cat did not appear to have any analgesia
after either IV or OTM administration of methadone.
This type of variation in response to administration of
methadone and other opioids has been observed previ-
ously in cats.3,6,8,10,16,34,35 While sex, genetics, and testing
modalities may all influence results of analgesic testing,
it is also possible that euphoric behaviors interfered
with our ability to properly apply the stimulus and to
interpret the responses. Similar challenges were previ-
ously reported in a study11 of buprenorphine adminis-
tration in cats.
Despite individual variations in response, our re-
sults suggest that, at the described doses, IV and OTM
administration of methadone were associated with anti-
nociceptive effects and behavioral changes (sedation
and euphoria) but had little to no influence on measured
physiologic parameters. Plasma drug concentrations did
not consistently parallel analgesic or sedative effects.
a. Methadone Hydrochloride Injection USP, 10 mg/mL, Xenodyne
Pharmaceuticals Inc, Newport, Ky.
b. Hydrion single-roll pH-paper dispenser, Micro Essential Labo-
ratory Inc, Brooklyn, NY.
c. Attane, Minrad Inc, Bethlehem, Pa.
d. 811-B ultrasonic Doppler flow detector, Parks Medical Electron-
ics Inc, Aloha, Ore.
e. Long-term MILACATH kit, MILA International Inc, Erlanger, Ky.
f. BD Insyte IV Catheter, 1.00 inches (0.9 X 25 mm), Becton Dick-
inson Infusion Therapy Systems Inc, Sandy, Utah.
g. MB International microcapillary centrifuge, International
Equipment Co, Needham, Mass.
h. Reichert TS Meter, Reichert Analytical Instruments, Depew, NY.
i. MT-H19 quick-read digital thermometer, Walgreens, Deerfield, Ill.
j. Pain Diagnostics and Treatment Inc, Greatneck, NY.
k. Mama KR, Mich PM, Raske T, et al. Plasma concentration and
selected behavioral effects following intravenous and oral trans-
mucosal buprenorphine in dogs, in Proceedings. Annu Meet As-
soc Vet Anesth 2008;65.
l. Agilent 1200 Series, Agilent Technologies, Santa Clara, Calif.
m. CTC Analytics HTC PAL System, Leap Technologies, Carrboro,
NC.
n. Sunfire C18 column, 4.6 X 50-mm internal diameter, 5.0-µm
bead size, Waters Corp, Milford, Mass.
o. SecurityGuard C18 cartridge, 4 X 2.0-mm internal diameter,
Phenomenex, Torrance, Calif.
p. API 3200 triple quadrupole instrument, Applied Biosystems
Inc, Foster City, Calif.
q. SAS System for Windows, version 9.2, SAS Institute Inc, Cary, NC.
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