STUDY PROTOCOL
EVALUATION OF ANALGESIC POTENTIAL OF TEST COMPOUND(S) USING FORMALIN TEST IN EXPERIMENTAL ANIMALS
1.0 INTRODUCTION:
The most predictive among the all models of acute pain is the Formalin Test. This is the widely used experimental model, a 5% solution of formaldehyde is injected subcutaneously to mouse or rat paw to produce a biphasic pain response over a test period of 60 min. The formalin test in mice is a valid and reliable model of nociception and is sensitive for all classes of analgesic drugs. The noxious stimulus is an injection of dilute formalin (1% in saline) under the skin of the dorsal surface of the right hindpaw.
2.0 TEST SYSTEM DETAILS:
Species : Mus musculus
Strain : Swiss albino
Age : 6-8 weeks
Body Wight : 20-25 g
Sex : Male
No. of animals : 8 /Group
3.0 ALLOCATION OF GROUPS:
|
Groups |
Treatment |
Dose; ROA |
No. of Animals |
|
G1 |
Normal
Control |
Normal
saline or 0.25% Na-CMC |
8 |
|
G2 |
Reference
Drug- Indomethacin |
10
mpk; p.o. |
8 |
|
G3 |
Test
Compound- I |
X
mpk; p.o. |
8 |
|
G4 |
Test
Compound- I |
XX
mpk; p.o. |
8 |
|
G5 |
Test
Compound- II |
X
mpk; p.o. |
8 |
|
G6 |
Test
Compound- II |
XX
mpk; p.o. |
8 |
Extra 5 animals will be taken, considering animal screening based on the basal latency time
4.0 METHOD:
· Animals shall be procured from the CPCSEA authorized vendor.
· Animals shall be quarantined for 1 week as per the in house SOP.
· Healthy animals will be selected, randomized based on body weight and divided into 6 different groups consisting of 8 animals each.
· Group G1 animal will be treated as normal control and treated with normal saline or Na- CMC.
· Animals of group G2 will be treated orally with Indomethacin at the dose of 10 mg/kg, p.o.
· Group G3, G4, G5 and G6 animals will be treated with test compound at different dose levels.
· The formalin test is an acceptable and reliable model of nociception that generates two separate phases of increased licking activity that can be recognized to different nociceptive mechanisms.
· The initial licking stage lasts for the first five minutes and a late phase takes from 15 to 45 minutes after the injection of formalin.
· As previously defined, formalin (20 μL of 2.5% solution) will be injected subcutaneously into the dorsal surface of the right hind paw after 30 min of drug treatment.
· The animals will be then placed under a glass funnel on a glass surface; a mirror was angled at 45 degrees.
· The pain response time (licking time) will be calculated in two periods: 0 to 5 min, the first phase (caused by direct motivation of the nociceptors), and 15 to 45 min, the second phase (inflammatory pain produced by release of inflammatory mediators).
5.0 END POINT PARAMETER(S):
· Paw licking time
· % Activity
6.0 REFERENCE(S):
5.1 Masoume Rezaee-Asl, Mandana Sabour, Vahid Nikoui, Sattar Ostadhadi, and Azam Bakhtiarian. The Study of Analgesic Effects of Leonurus cardiaca L. in Mice by Formalin, Tail Flick and Hot Plate Tests. International Scholarly Research Notices Volume 2014 (2014), Article ID 687697, 5 pages
5.2 D. Dubuisson and S. G. Dennis, “The formalin test: a quantitative study of the analgesic effects of morphine, meperidine, and brain stem stimulation in rats and cats,” Pain, vol. 4, no. 2, pp. 161–174, 1978.
5.3 Bao F, Wu P, Xiao N, Qiu F, Zeng QP. Nitric
oxide-driven hypoxia initiates synovial angiogenesis, hyperplasia and
inflammatory lesions in mice. PloS one. 2012 Mar 30;7(3):e34494.
END OF DOCUMENT
You may like to read theses links:
6.EVALUATION OF ANALGESIC POTENTIAL OF TEST COMPOUND(S) USING HOT PLATE TEST IN EXPERIMENTAL ANIMALS
Hi. I’m Writer of Researchsop.com. ’ ’ Please share these SOPs to all concern pharma people for their development. I like to fullfill the need of curious people. These things inspire me to make things looks better.
0 comments:
Post a Comment