Bio-Analytical SPE Method: Empagliflozin and Sitagliptin
Estimation of Empagliflozin and Sitagliptin from human plasma samples using LC/MS-MS
Empagliflozin
Empagliflozin is an anti-diabetic medication. It works by removing excess sugar from your body through urine.
Sitagliptin
Sitagliptin is used in the treatment of type 2 diabetes mellitus. It is used along with diet and exercise to improve blood sugar control in adults with type 2 diabetes.
Dose, formulation and route of administration
10 mg/100mg tablets, Oral administration
Method Details
Drug Information
| Drug | Chemical Structure | Molecular Weight | LLOQ & ULOQ |
|---|---|---|---|
| Empagliflozin | Chemical structure | 450.91 g/mole | 1-1000 ng/ml |
| Sitagliptin | Chemical structure | 407.31 g/mole | 2-1000 ng/ml |
Chromatographic Conditions
| Parameters | Details |
|---|---|
| Mobile phase | 5 mM ammonium acetate : Acetonitrile (35:65 v/v) |
| Column | 3 μm, HyPURITY C18 column (100 × 4.6 mm) |
| Flow rate | 0.6 mL/min |
| Column temp | 45°C |
Mass Spectrometry Conditions
| Parameters | Details |
|---|---|
| Mass spec | API-4000 |
| Empagliflozin m/z | 451.04 → 71.04 |
| Empagliflozin D4 m/z | 455.04 → 75.04 |
| Sitagliptin m/z | 408.1 → 235.1 |
| Sitagliptin D4 m/z | 412.1 → 239.1 |
| Ion source | ESI spray |
| Mode | Positive ionization mode |
Sample Preparation (SPE, Affinity HLB 25mg, 1 cc cartridges)
| Step | Procedure |
|---|---|
| Sample pretreatment | Plasma + drug + IS- Add 500 uL 0.1% formic acid and shake well. Centrifuge, if needed before application on SPE |
| Condition | 1 ml Methanol |
| Equilibration | 1 ml Water |
| Sample Loading | Load supernatant sample |
| Washing | Wash-1: 1 ml water Wash-2: 10% to 30% Methanol in water (Optimize this step for best results) |
| Elution | 500 to 1 ml mobile phase |
| Direct Injection | 10 uL |