DTDPA-OPA联合柱前衍生化HPLC测定普卡那肽原料药中的氨基酸比值

Determination of Amino Acid Ratios in Plecanatide by DTDPA-OPA Combined with Pre-column Derivatization HPLC

采用HPLC法,以二硫代二丙酸(DTDPA)和邻苯二甲醛(OPA)联合作为柱前衍生化试剂,建立了普卡那肽原料药中氨基酸比值的测定方法。结果显示,普卡那肽中各氨基酸的定量限为0.0069~0.0127μmol/mL,检测限为0.0035~0.0063μmol/mL,在定量限至测定浓度的200%范围内,线性关系良好(r≥0.9996);溶液稳定性RSD(n=9)≤4.058%;氨基酸比值的精密度RSD(n=12)≤2.41%;平均回收率(n=9)在97.32%~101.96%,RSD≤1.22%;检测条件发生微小变化时,氨基酸比值的RSD(n=16)≤1.61%。该法专属性强、准确、高效,可用于普卡那肽氨基酸比值的测定。

An pre-column derivatization HPLC method with dithiodipropionic acid(DTDPA)and o-phthalaldehyde(OPA)was established to determine the amino acid ratios in plecanatide.The results showed that the quantitative limits were 0.0069-0.0127μmol/mL,the detection limits were 0.0035-0.0063μmol/mL.The linearity and solution stability of all amino acids were good with minimum r of 0.9996 and the maximum RSD(n=9)of 4.058%.The precision was good with the maximum RSD(n=12)of 2.41%.The mean recoveries(n=9)were 97.32%-101.96%,the maximum RSD was 1.22%.In case of detection parameters slight change,the durability was good with the maximum RSD(n=16)of 1.61%.The established method is specific,accurate and efficient,which can be used for determination of the amino acid ratios of plecanatide.