Screening of Local

Anaesthetics
Dr. Advaitha
M.V
KSHEMA, Mangalore

Types of local Anaesthesia in

Preclinical models
1.
2.
3.
4.
5.

Conduction Anaesthesia
Infiltration Anaesthesia
Surface Anaesthesia
Epidural Anaesthesia
Spinal Anaesthesia

Conduction Anaesthesia in the

Sciatic Nerve of the Rat

Definition:
It is a type of regional anesthesia.
local anesthetic solution is injected to
nerves to inhibit nerve transmission.
It includes
Spinal, epidural,
Nerve block, and field block anesthesia,
but not topical anesthesia.
But for all practical Purposes, Here it only
means Nerve Block

 Wistar or Sprague Dawley rats

Weight : 125 to 175 g

Procedure:
Grasp the base of the tail and thoracic
cage.
The animal is suspended in a prone
position.
A hind limb is extended to its full length.
Site of injection : area under the skin
at the junction of the biceps femoris and
the gluteus maximus muscles.

 The sciatic nerve is blocked in the

midthigh region with 0.2 ml of the
drug solution.
(use a 24- to 25-gauge needle
attached to a 0.25 ml tuberculin
syringe).
The other leg is used for the control
drug.

 Immediately after the injection,

repeated check of the digit of the foot,
the walking behavior.

is observed.

In the normal foot, the digits are wide

apart.

In the blocked leg the digits of the foot

are close together.

 The successful block is evidenced by

Dragging of the leg
Inability of the animal to use the leg in
walking up the inclined wire mesh
cover of the cage.
Lastly,
The time of recovery :
After the time of block for each leg is
noted, each animal is examined every
5 to 10 min.

Similar Experiments of conduction

Anaesthesia :
Conduction Anaesthesia in Sciatic
nerve of Frog.
Conduction anaesthesia on Mouse
Tail.

Infiltration Anaesthesia in Guinea

Pigs Wheals
BASED ON
works of
Mc Intyre & sievers(1937)
Bulbring & wajda (1945)
This method is one of the standard
operating procedure for testing local
anesthesia
Adult Guniea Pigs
Weight: 250 300 g

 Procedure:
The sensitivity of the skin is greatest
in the midline.
Also, It is slightly more in the front
than in the back area.
So, each concentration of a local
anesthetic must be tested in both
areas.

 One day preceding the experiment

two areas of 45 cm diameter are
shaved.
This produces certain amount of
sensitivity which disappears
overnight

 The doses of local anesthetics are

injected intracutaneously in 0.1 ml
saline.
The size of the wheal is marked with
ink.
One side is used for the test
preparation, the other side for the
standard
(e.g 1%butanilicaine) .

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 The reaction to pin prick is tested 5

min after injection in the following
way- Firstly, normal reaction to a prick
applied outside the wheal is seen.
Then six pricks are applied inside the
wheal.
The pricks are applied at intervals of
about 3 5 s.

 Six pricks are applied every 5 min for

30 min.
Now, the number of pricks to which
the guinea pig fails to react inside
the wheal during 30 min noted.
This gives an indication of degree of
anesthesia.

Surface Anaesthesia on the Cornea

of Rabbits
Animals: Albino Rabbits
Weight: 2.5 3 kg

Procedure:
The upper and lower eyelashes are
carefully clipped.
The conjunctival sac of one eye is held
open, forming a pocket.
Apply 0.5 ml of a solution of the anesthetic
into the conjunctival sac for 30 s.

(use, 1 ml syringe with a 22-gauge

needle)

 1 ml of the standard is applied to the

other eye ( 0.1% solution of
tetracanine hydrochloride).

 Effective local anesthetics abolish

the corneal reflex (blinking) elicited
by any touch of the cornea.
The test is started 5 min after
application of the drug.
It is repeated every 5 min until
effect subsides and blinking occurs
again.
The time between disappearance
and reappearance of the corneal

Method of irritating cornea

For quantitative purpose the
irritation, von frey hairs has been
widely recommended.
An equine(HORSE) hair bending at a
load of 230 mg is attached
perpendicularily to a glass rod.
Within 25 sec, the cornea is touched
100 times.

 The summation of many stimuli

applied this way gives better result
than a single touch with the glass
rod (Ther and Mugge1953)

Von Frey hairs. Through the use of a camel hair,

various levels of pressure are applied to the skin
and pressure is calculated. The hair is mounted
inside a tube. Calculations are made respective to
how much the hair is bent during the examination

Today, nylon fibers are used to perform the

same task.

Similar experiments:
Abolition of Sneezing Reflex in Rabbits
New Zealand Albino rabbits weighing
3kg are used.
The test solution is applied to the
mucous membrane of one nostril and
standard to other.

 After 2-5 min mucosa is stimulated with

a lead pencil.
Loss of sneezing reflex is considered as
the sign of complete local anaesthesia.
Test is repeated at 3,5,10 and 15min
and continued every 5 min until reflex
reappears.

Epidural Anesthesia in
Rabbits
Animals: Adult Rabbit

Weight: 2.5 3 kg

 Procedure:
The rabbit is carefully restrained by
an assistant.
Palpate the midline at L7 spine and
the depression over the lumbosacral
fossa.

The Rabbit Vertebrae column consists of 7

Cervical Vertebrae (As in all mammals), 1213 Thoracic Vertebrae, 7 Lumbar, 4 Sacral,
and a variable amount of caudal vertebrae
(usually 16)

 At this point, through a short bevel

1.5 cm, 20 gauge spinal needle to
be introduced.
This is at right angles to the skin in
the midline with the bevel aligned
longitudinally

 After passage through the skin,

only minor resistance would be felt
until the ligamentum flavum is
reached.
When passing through the ligament, a
definite pop is felt and resistance to
advancement of the needle is lost.
The stylet would be then withdrawn
and the hub inspected for the presence
of blood or cerebrospinal fluid.

If absent, the needle should be rotated

through 90 to direct the bevel
caudally.

A 1.0 ml syringe to be attached and

0.1 ml of air injected.

A syringe containing the desired dose

of the local anesthetic is injected over
a 5 10 s period.

 The effectiveness is assessed by

(1) Sensory loss,
(2) loss of weight-bearing ability, and
(3) flaccid paresis.

Spinal Anaesthesia in Rats

Animals: Sprague-Dawley rats

Weight: 75- 100 g

Procedure:
A 30-gauge needle is attached to a 25l
Hamilton syringe.
This is introduced on one side of the L5
or L6 spinous process at an angle of
about 20.
The needle is advanced to the groove
between the spinous and transverse
processes.

 Then moved forward to intervertebral

space at an angle of about 10.
Correct placement of the needle is
indicated by an arching of the tail.
Drugs are dissolved in saline or water
and administered in a volume of 5
l.

 Antinociception is determined by tail

flick assay.
(by placing the tail under a focused
radiant heat source on an
analgesiometer.)

The reaction time is noted in each

case.

References:
Drug Discovery and Evaluation:
Pharmacological Assays, 3rd Edition;
by H. Gerhard Vogel.

Thank You