Femtomole quantitation of 7-ethyl-10-hydroxycamptothecine (SN-38) in plasma samples by reversed-phase high-performance liquid chromatography
Femtomole quantitation of 7-ethyl-10-hydroxycamptothecine (SN-38) in plasma samples by reversed-phase high-performance liquid chromatography
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DOI：
10.1006/abio.1999.4001
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发表时间：
1999-04-10
影响因子：
2.9
通讯作者：
Sparreboom, A
中科院分区：
文献类型：
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作者：
de Bruijn, P;de Jonge, MJA;Sparreboom, A
7-乙基-10-羟基喜树碱（SN-38）是拓扑异构酶I抑制剂和抗癌药伊立替康（CPT-11）的活性代谢产物。在这里，我们提出了一种新的和灵敏的反相高效液相色谱法测定人血浆样品中的SN-38。样品预处理包括用2 mL乙腈沉淀1 mL样品的蛋白质，然后用5 mL氯仿进行一步溶剂萃取，喜树碱用作内标。在填充有Hypersil ODS材料（100 × 4.6 mm i.d.，5 μ m P.S.），用含有10 mM四丁基硫酸铵的乙腈：0.1 M乙酸铵（23：77，v/v），pH 5.3（盐酸）的混合物进行等度洗脱。分别在380和556 nm的激发和发射波长下监测色谱柱流出物。该方法的定量限为低飞摩尔水平（类似于8.4 fmol;相当于5 pg/mL），标准曲线在近三个数量级上呈线性。试验内精密度为
7-Ethyl-10-hydroxycamptothecine (SN-38) is the active metabolite of the topoisomerase I inhibitor and antineoplastic agent, irinotecan (CPT-11). Here, we present a new and sensitive reversed-phase high-performance liquid chromatographic method for the determination of SN-38 in human plasma samples. sample pretreatment involves a protein precipitation of 1-mL samples with 2 mL of acetonitrile, followed by a one-step solvent extraction with 5 mL of chloroform, with camptothecine used as internal standard. Chromatographic separation was achieved on an analytical column packed with Hypersil ODS material (100 x 4.6 mm i.d., 5 mu m P.S.), and isocratic elution with a mixture of acetonitrile:0.1 M ammonium acetate containing 10 mM tetrabutylammonium sulfate (23:77, v/v), pH 5.3 (hydrochloric acid). The column effluent was monitored at excitation and emission wavelengths of 380 and 556 nm, respectively. The limit of quantitation of the method presented was at the low femtomole level (similar to 8.4 fmol; equivalent to 5 pg/mL), with the standard curves being linear over nearly three orders of magnitude. Intraassay precision was