HPLC法测定重组人甲状旁腺素1-34肽含量及有关物质

HPLC determination of peptide content and related substances of recombinant human parathyriod hormone 1-34

目的:用高效液相色谱法测定重组人甲状旁腺素1-34原液和制剂的肽含量及有关物质。方法:采用 Zorbax 300SB-C_(18)色谱柱(250mm×4.6mm,5μm),以流动相 A:水-乙腈-三氟乙酸(990:10:1),流动相 B:水-乙腈-三氟乙酸(10:990:1)为流动相,梯度洗脱,流速1.0mL·min^(-1),检测波长215nm,柱温37℃。结果:重组人甲状旁腺素1-34浓度在26.25～841.5μg·mL^(-1)范围内,与峰面积的线性关系良好(r=0.9999),最低检测限为131ng,高、中、低3种浓度的平均回收率范围为99.5%～100.6%,RSD 为0.52%～0.72%(n=3)。结论:方法简便,快速,准确,可靠,可作为本品的质量控制方法。

Objective： To establish an HPLC method for the determination of peptide content of recombinant human parathyriod hormone 1 -34 （rhPTH 1 -34 ） and its related substances in its law materials and preparations. Methods：The separation was performed on a Zorbax 300SB -C18column（250 mm ×4. 6 mm,5 μm） using gradient elution with solution A of water- acetonitrile -trifluoroacetic（990： 10：1 ） and solution B of water - acetonitrile - trifluoroacetic（ 10： 990：1 ） as mobile phase;The flow rate was 1.0 mL · min^-1 The detection was set at 215 nm and the column temperature was maintained at 37℃. Results：The calibration curve was linear in the concentration range of 26. 25 - 841.5 μg · mL^- 1 with correlation coefficient of 0. 9999. The detection limit was 131 ng. The average recovery was from 99. 5% to 100. 6% for three different levels,RSD was from 0. 52% to 0. 72% （n =3）. Condusion：This method is simple, rapid, accurate, reliable and suitable for the quality control of this drug.