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This design provides: ✓ Enhanced retention of strong basic compounds via cation-exchange mechanism ✓ Reverse-phase retention of neutral compounds ✓ Additional selectivity for polar molecules, enabling separation of isomers and structurally related compounds ✓ Flexible mobile phase compatibility, suitable for LC-MS, preparative chromatography, and low UV detection ⚙️ Typical HPLC Method Parameters ✅ Column: Primesep 200, 3.2 × 150 mm, 5 µm, 100 Å, dual-ended ✅ Mobile Phase: MeCN – 20% ✅ Buffer: Gradient ammonium formate pH 3.0, 20–60 mM over 10 min ✅ Flow Rate: 0.5 mL/min ✅ Detection: LC-MS, ESI, GHK (free peptide, [M+H]+) 341 ✅ LOD: 30 ppb *LOD was determined for this specific instrument, method, and analyte