Due to the tidal schedules and the fact that these species are in the inter-tidal zone, the C. coronatus specimens were collected from Zeyton Park of Qeshm Island (location: N 26055.631; E 056015.209).
Extraction
conopeptide extraction was performed by venom ducts isolation from the cone snails on ice, then cold water and acetonitrile 40%, which completely deoxygenation was added and slowly homogenized in a homogenizer (model Silent crasher, Heidolph, Germany), in ꒼ 16000 rpm for 5 minutes. Then, for 20 minutes at 4 ° C and ꒼ 10000 g were centrifuged. The supernatant containing the conopeptides had been lyophilized at the freeze dryer (Model Christ, 2 alpha, Germany), slowly for 24 hours at -56 0C (Tayo et al, 2011).
Analgesic test
The …show more content…
The number of mice in each experimental group was 7. This experiment was performed in a box with dimensions of 30 × 30 × 30. Initially, 200 ml of the extract and purified fractions at a dose 0/5 mg/kg were injected IP on mice. After about one hour, 20 ml of formalin% 2/5 subcutaneously injected into right paw of the mice, then for one hour pain response was assessed. Flinching and licking number were considered as pain response (Green et al., 2007; Han et al., …show more content…
Peak analysis was performed on Chromgate v 3.3.2 software. The diluted fraction was injected to appropriate concentration on Analytical C18 column (Coulter model, made in America). This column has the capacity to accept 165 µg. The column had the following characteristics (particle size: 5 µ, pore size: 300 Å, column size: 4/6 ꒼250mm). For conopeptide isolation, deionized water and acetonitrile, both containing 0/05%TFA with program (0 to 90% acetonitrile) for 90 minutes at a rate of 1 ml/min was
Extraction
conopeptide extraction was performed by venom ducts isolation from the cone snails on ice, then cold water and acetonitrile 40%, which completely deoxygenation was added and slowly homogenized in a homogenizer (model Silent crasher, Heidolph, Germany), in ꒼ 16000 rpm for 5 minutes. Then, for 20 minutes at 4 ° C and ꒼ 10000 g were centrifuged. The supernatant containing the conopeptides had been lyophilized at the freeze dryer (Model Christ, 2 alpha, Germany), slowly for 24 hours at -56 0C (Tayo et al, 2011).
Analgesic test
The …show more content…
The number of mice in each experimental group was 7. This experiment was performed in a box with dimensions of 30 × 30 × 30. Initially, 200 ml of the extract and purified fractions at a dose 0/5 mg/kg were injected IP on mice. After about one hour, 20 ml of formalin% 2/5 subcutaneously injected into right paw of the mice, then for one hour pain response was assessed. Flinching and licking number were considered as pain response (Green et al., 2007; Han et al., …show more content…
Peak analysis was performed on Chromgate v 3.3.2 software. The diluted fraction was injected to appropriate concentration on Analytical C18 column (Coulter model, made in America). This column has the capacity to accept 165 µg. The column had the following characteristics (particle size: 5 µ, pore size: 300 Å, column size: 4/6 ꒼250mm). For conopeptide isolation, deionized water and acetonitrile, both containing 0/05%TFA with program (0 to 90% acetonitrile) for 90 minutes at a rate of 1 ml/min was