Leuprolide

Molecular formula: C 59 H 84 N 16 O 12 Molecular weight: 1209.4

CAS Registry No.: 53714-56-0 (leuprolide), 74381-53-6 (leuprolide acetate)

SAMPLE Matrix: formulations Sample preparation: Mix sample with 2 mg/mL ethyl p-hydroxybenzoate in MeOH so as to give 100 |xg/mL leuprolide acetate and 150 |xg/mL ethyl p-hydroxybenzoate in saline, inject a 20 jxL aliquot. HPLC VARIABLES Column: 150 X 4-4.5 5 |xm octadecylsilane (IBM5 Bio-Rad, or Nucleosil) Mobile phase: MeCN: buffer 23:77 Saturate mobile phase with silica by slurrying with Alltech Adsorbosil then filtering (0.4 |xm). (Buffer was 87 mM (NH4)H2PO4 adjusted to pH 6.5 with ammonium hydroxide.) Flow rate: 2 Injection volume: 20 Detector: UV 220 CHROMATOGRAM Retention time: 19 (leuprolide acetate) Internal standard: ethyl p-hydroxybenzoate (7) OTHER SUBSTANCES Simultaneous: degradation products KEYWORDS injections; stability-indicating REFERENCE
Sutherland, J.W.; Menon, G.N. HPLC of leuprolide acetate in injectable solutions. J.Liq.Chromatogr., 1987, 10, 2281-2289

SAMPLE Matrix: solutions HPLCVARIABLES Column: 200 X 3 Spherisorb S5ODS-2

Mobile phase: Gradient. A was 0.05% phosphoric acid containing 0.5% (NH4)2SO4. B was MeCN. A:B from 82:18 to 64:36 over 25 min, maintain at 64:36 for 2.5 min, return to initial conditions over 1 min, re-equilibrate for 6.5 min. Alternatively, isocratic MeCN: 0.05% phosphoric acid containing 0.5% (NH4J2SO4 24:76. Flow rate: 0.5 Detector: UV 210 CHROMATOGRAM Retention time: 20 (gradient), 13 (isocratic) OTHER SUBSTANCES Simultaneous: buserelin, deslorelin, gonadorelin, goserelin, nafarelin KEYWORDS comparison with capillary electrophoresis REFERENCE
Corran, RH.; Sutcliffe, N. Identification of gonadorelin (LHRH) derivatives: comparison of reversedphase high-performance liquid chromatography and micellar electrokinetic chromatography. J.Chromatogr., 1993, 636, 87-94