Analytical Test
Identification
Analytical Technique
HPLC
System Suitability
Internal standard method with progesterone as internal standard.
| Experimental Setting | Value |
|---|---|
| HPLC | Agilent 1100 series pump, solvent degasser, autosampler, DAD detector |
| Column | 150 x 4.6 mm, 5 μm Hichrom C18 |
| Mobile Phase | 65:35 v/v methanol/acetate buffer (0.02 M sodium acetate adjusted to pH 5.0 with glacial acetic acid) |
| Flow Rate | 1.0 mL/min |
| Column Temperature | 35° C |
| Detection λ | 245 nm |
| Injection Volume | 20 μL |
| Standard Stock Solutions | 0.5 mg MPA and progesterone (PG) in mobile phase. Working solutions prepared by diluting stock solutions with the mobile phase to contain 100-300 μg/mL of MPA and 150 μg/mL PG as internal standard. Megestrol acetate (MGA) added to the standard solution for a purity test of bulk drug. Resolution should be not less than 1.5. |
| Sample Preparation | Injectable suspensions from 20 vials were pooled and shaken on mechanical shaker for 30 min. A portion of the suspension equivalent to 50 mg MPA was weighed and transferred to a 50 mL volumetric flask. 10 mL methanol added. Shaken for 10 min, then adjusted to volume and mixed well. A 2 mL aliquot is transferred to a 10 mL volumetric flask containing 1500 μg internal standard. Sample diluted to volume with mobile phase and filtered through a 0.45 μm nylon syringe filter. |
References