Systemic injection test
1. Biological Effect: In Vivo Tests
2. Turnaround time: 6 weeks (after approval from Local Ethics Committee)
3. Sample Requirements:
USP
to be individually determined;
4. Animal Quantity: 40 mice
USP <88>
This test is designed to evaluate systemic responses to the extracts of materials under test following injection into mice.
Test Animals
Use healthy, not previously used albino mice weighing between 17 and 23 g. For each test group use only mice of the same source. Allow water and food, commonly used for laboratory animals and of known composition, ad libitum.
Procedure
Inject each of the five mice in a test group with the Sample or the Blank, except to dilute each g of the extract of the Sample prepared with Polyethylene Glycol 400, and the corresponding Blank, with 4.1 volumes of Sodium Chloride Injection to obtain a solution having a concentration of about 200 mg of polyethylene glycol per mL.
| EXTRACT OR BLANK | DOSE PER KG | ROUTE | INJECTION RATE (μL per sec) |
|---|---|---|---|
| Sodium Chloride Injection | 50 mL | intravenous | 100 |
| 1 in 20 solution of Alcohol in Sodium Chloride Injection | 50 mL | intravenous | 100 |
| Polyethylene Glycol 400 | 10 g | intraperitoneal | – |
| Vegetable Oil | 50 mL | intraperitoneal | – |
Observation
Observe the animals immediately after injection, again 4 h after injection, and then at least at 24, 48, and 72 h. If during the observation period none of the animals treated with the extract of the Sample shows a significantly greater biological reactivity than the animals treated with the Blank, the Sample meets the requirements of this test. If two or more mice die, or if abnormal behavior such as convulsions or prostration occurs in two or more mice, or if a body weight loss greater than 2 g occurs in three or more mice, the Sample does not meet the requirements of the test. If any animals treated with the Sample show only slight signs of biological reactivity, and not more than one animal shows gross symptoms of biological reactivity or dies, repeat the test using groups of 10 mice. On the repeat test, all 10 animals treated with the Sample show no significant biological activity above the Blank animals during the observation period.
