Tian Qingguo, Rogness Juan, Meng Min, Li Zheng
Bioanalysis, Arrowhead Pharmaceuticals, Inc., 502 S. Rosa Road, Madison, WI 53719, USA.
Bioanalytical lab, Covance Laboratories, Inc., 1121 East 3900 South, Salt Lake City, UT 84124, USA.
Bioanalysis. 2017 Jun;9(11):861-872. doi: 10.4155/bio-2017-0017. Epub 2017 Jun 15.
Toxicokinetic and pharmacokinetic studies of therapeutic oligonucleotides require validated bioanalytical methods for sensitive and specific quantification of oligonucleotide drug candidates in biological samples.
A peptide nucleic acid (PNA) hybridization-based HPLC-fluorescence assay was developed and validated for quantification of Arrowhead Pharmaceuticals' proprietary siRNA in rat plasma samples via hybridization and anion-exchange-HPLC (AEX-HPLC) with fluorescence detection.
The validated method provided a sensitive and selective approach for quantification of siRNA in biological samples at a linear quantitation range of 1-1000 ng/ml. The assay requires only 25 μl of plasma sample and shows excellent accuracy and precision even without using an internal standard, providing a useful quantification method for siRNA determination in biological matrix with limited sample volume.
治疗性寡核苷酸的毒代动力学和药代动力学研究需要经过验证的生物分析方法，以灵敏且特异的方式定量生物样品中的寡核苷酸候选药物。
开发并验证了一种基于肽核酸（PNA）杂交的HPLC-荧光分析法，用于通过杂交和阴离子交换-HPLC（AEX-HPLC）及荧光检测，定量大鼠血浆样品中箭头制药公司的专利小干扰RNA（siRNA）。
经过验证的方法为定量生物样品中的siRNA提供了一种灵敏且具选择性的方法，线性定量范围为1-1000 ng/ml。该分析方法仅需25 μl血浆样品，即使不使用内标也显示出优异的准确度和精密度，为在样品量有限的生物基质中测定siRNA提供了一种有用的定量方法。
