Goodyear N, Napoli K L, Murthy J N, Kahan B D, Soldin S J
Department of Laboratory Medicine, Children's National Medical Center, Washington, DC 20010, USA.
Clin Biochem. 1996 Oct;29(5):457-60. doi: 10.1016/0009-9120(96)00071-9.
To develop a radioreceptor assay (RRA) for sirolimus (rapamycin, RAPA).
A direct methanol extraction was used to prepare 45 patient samples for the RRA. Results were compared to the results obtained previously using high-performance liquid chromatography (HPLC). Between-run precision, recovery, and drug interference studies were also performed.
The RRA is sensitive to 1.0 microgram/L RAPA equivalents in whole blood. Comparison with HPLC yielded a correlation coefficient for 45 patient samples of 0.977. Between-run precision at 2.5, 7.5, 12.5, and 20 micrograms/L showed coefficients of variation (CVs) of 12.9, 9.2, 8.5, and 5.9%, respectively. Recoveries from the extraction procedure were 93% at 7.5 micrograms/L and 103% at 12.5 micrograms/L. Drug interference studies showed no interference in the RRA by cyclosporine (CsA), dexamethasone, prednisone, or methotrexate.
We have demonstrated that the RRA for RAPA correlates well with HPLC, and has excellent precision and recovery. The procedure is far less time-consuming and complex than HPLC and has potential for automation.
开发一种用于西罗莫司（雷帕霉素，RAPA）的放射受体分析法（RRA）。
采用直接甲醇萃取法为RRA制备45份患者样本。将结果与先前使用高效液相色谱法（HPLC）获得的结果进行比较。还进行了批间精密度、回收率和药物干扰研究。
RRA对全血中1.0微克/升的RAPA等效物敏感。与HPLC比较得出，45份患者样本的相关系数为0.977。在2.5、7.5、12.5和20微克/升时的批间精密度分别显示变异系数（CV）为12.9%、9.2%、8.5%和5.9%。萃取过程在7.5微克/升时的回收率为93%，在12.5微克/升时为103%。药物干扰研究表明，环孢素（CsA）、地塞米松、泼尼松或甲氨蝶呤对RRA无干扰。
我们已证明，RAPA的RRA与HPLC相关性良好，具有出色的精密度和回收率。该方法比HPLC耗时少得多且更简单，具有自动化潜力。
