Perform an HPLC injection accuracy test.

ENVIRONMENT

 Alliance HPLC System

 2695 Separations Module

 2695D Dissolution Separations Module

 717plus Autosampler

PROCEDURE

Injection Volume Accuracy when using the standard 250µL syringe

Note: To improve the accuracy of this test, handle the vial with forceps to ensure that

the vial is as clean as possible.

1. Fill a standard vial with methanol or water.

2. Seal the vial with a Teflon Septum and cap.

3. Zero the analytical balance, carefully weigh the vial, and record the weight.

4. Place the vial in the carousel and set the flow rate to 1.0mL/min.

5. Program the system to make six 50µL injections from the vial. (Use a run time of

approximately one minute or less)

6. Weigh the vial after the last injection and record that weight.

Using one of the following equations (depending on the solvent used), calculate the

average amount injected per injection.

((W1-W2) /6) x (1/density of solvent) = µL injected

Using Methanol

Density of Methanol is 0.7890

((W1-W2) /6) x 1.2687 = µL injected

Using Water

Density of Water is 1.0000

((W1-W2) /6) x 1.0000 = µL injected

RESULTS: 50µl +/- 2µL