HPLC测定鲑鱼降钙素磷脂复合物的含量

Determination of Salmon calcitonin-phospholipid complex by HPLC

目的建立鲑鱼降钙素磷脂复合物的含量测定方法。方法采用HPLC法，Vydac218TP54C18色谱柱（250mm×4．6mm，5μm，300A），流动相为0．02mol·L-1四甲基氢氧化铵缓冲液（磷酸调pH2．5）-乙腈（A：9：1，B：2：3），梯度洗脱程序为：0—21min、65％～43％A，21．01～24min、65％A，流速1．0mL·min-1，检测波长220nm，柱温40℃，进样量20μL。结果鲑鱼降钙素检测浓度的线性范围为4．49—89．87μg·mL-1（r=0．9995），平均回收率为99．56％（RSD：1．48％）。结论所用方法准确可靠、重复性好，可用于鲑鱼降钙素磷脂复合物的含量测定。

OBJECTIVE To establish a content determination method for Salmon calcitonin - phospholipid complex. METHODS The chromatographic separation was performed on a Vydac 218TP54 C18 column （250 mm × 4.6 mm,5 μm,300 A） with a mobile phase of 0.02 mol L-1 tetramethyl ammonium hydroxide buffer （adjusted with phosphoric acid to a pH of 2.5 ） -acetonitrile （A ：9 ： 1, B： 2 ： 3）. The chromatograph was programmed as follow：0 -21 min,65% -43% A,21.01 -24 min,65% A. The flow rate was 1 mL min-1 and detection wavelength was 220 nm. The column temperature was maintained at 40 ℃ and the inject volumes was about 20 μL. RESULTS The linear ranges was in the range of 4.49 - 89.87 μg. mL-1 （ r = 0. 9995 ）. The average recovery was 99.56% （ RSD = 1.48% ）. CONCLUSION The method is accurate, reproducible,and can be used as a content determination method for Salmon calcitonin - phospholipid complex.