De Ceballos M L, Taylor M D, Jenner P
Parkinson's Disease Society Experimental Research Laboratory, King's College London, UK.
Neuropeptides. 1991 Nov;20(3):201-9. doi: 10.1016/0143-4179(91)90132-3.
A reverse-phase, high-performance liquid chromatographic (HPLC) method was employed to separate and characterise five neuropeptides from complex mixtures, with important advantages over methods employed earlier using combined HPLC-RIA studies. Peptides were separated using 0.5M pyridine-0.5M formic acid buffer, pH 4, containing propan-l-ol 14% (met-enkephalin, leu-enkephalin, neurotensin) or 20% (CCK-8-S, substance P) at a flow rate of 1.0 ml/min. Isocratic conditions, and volatile solvents, resulted in a highly reproducible method, producing samples in a form designed for subsequent RIA. The application and importance of the procedure is demonstrated by comparison of the measurements of apparent peptide levels in crude brain extracts with those of authentic peptides as determined after HPLC purification.
采用反相高效液相色谱（HPLC）法从复杂混合物中分离并鉴定了五种神经肽，与早期使用HPLC-RIA联合研究的方法相比具有重要优势。使用含14%丙醇（甲硫氨酸脑啡肽、亮氨酸脑啡肽、神经降压素）或20%丙醇（CCK-8-S、P物质）的0.5M吡啶-0.5M甲酸缓冲液（pH 4），以1.0 ml/min的流速分离肽段。等度条件和挥发性溶剂产生了一种高度可重复的方法，以适合后续放射免疫分析（RIA）的形式制备样品。通过比较粗脑提取物中表观肽水平的测量值与HPLC纯化后测定的真实肽的测量值，证明了该方法的应用和重要性。
