A high-performance liquid-chromatographic method for routine simultaneous determination of nicotine and cotinine in plasma.
A high-performance liquid-chromatographic method for routine simultaneous determination of nicotine and cotinine in plasma.
复制标题
常规同时测定血浆中尼古丁和可替宁的高效液相色谱方法。
DOI：
10.1093/clinchem/34.4.724
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发表时间：
1988
影响因子：
9.3
通讯作者：
J. F. Greden
中科院分区：
文献类型：
--
作者：
M. Hariharan;T. VanNoord;J. F. Greden
我们描述了一种快速、灵敏的常规同时测定1毫升血浆中尼古丁和可替宁的方法。用三氯乙酸脱蛋白后用二氯甲烷抽提10毫升螺帽特氟龙管，可消除乳状液形成。萃取物在30微升的流动相中蒸发和重建后，被注入等度高效液相色谱装置的反相C-18离子对色谱柱。在256 nm处监测吸光度。流动相为柠檬酸盐-磷酸盐缓冲液(每升30毫克)，每公升含有50毫升乙腈和1毫升庚烷磺酸钠。2-苯基咪唑为内标。尼古丁的检出限为1微克/L，可替宁为3微克/L。两种化合物的标准曲线在0-700微克/L范围内呈线性关系。尼古丁在0-100微克/L浓度范围内的平均变异系数为6.5%，可替宁在50-700微克/L浓度范围内的平均变异系数为4%。
We describe a rapid, sensitive method for the routine simultaneous determination of nicotine and cotinine in 1 mL of plasma. Extraction in 10-mL screw-capped Teflon tubes with methylene chloride after deproteinization with trichloroacetic acid eliminated emulsion formation. The extract, after evaporation and reconstitution in 30 microL of mobile phase, is injected into a reversed-phase C-18 ion-pair column of an isocratic high-performance liquid-chromatographic unit. Absorbance is monitored at 256 nm. The mobile phase is a citrate-phosphate (30 mmol each per liter) buffer mixture containing 50 mL of acetonitrile and 1 mmol of sodium heptanesulfonate per liter. 2-Phenylimidazole is the internal standard. The detection limit is 1 microgram/L for nicotine and 3 micrograms/L for cotinine. The standard curve is linear from 0 to 700 micrograms/L for both compounds. The average CV for nicotine in the concentration range 0-100 micrograms/L is 6.5%, and that for cotinine in the concentration range 50-700 micrograms/L is 4%.