使用反相高效液相色谱法优化犬血浆中番茄红素的测定
Optimized determination of lycopene in canine plasma using reversed-phase high-performance liquid chromatography.
作者信息
Vertzoni M V, Reppas C, Archontaki H A
机构信息
Laboratory of Biopharmaceutics and Pharmacokinetics, School of Pharmacy, National and Kapodistrian University of Athens, Panepistimiopolis, Athens 15771, Greece.
出版信息
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 May 5;819(1):149-54. doi: 10.1016/j.jchromb.2005.02.001.
An isocratic high-performance liquid chromatographic method with detection at 472 nm was developed, optimized and validated for the determination of lycopene in canine plasma. Ethyl-beta-apo-8'-carotenoate was used as internal standard. A Hypersil BDS RP-C18 column (150 mm x 4.6 mm), 5 microm particle size, was equilibrated with a mobile phase composed of acetonitrile and methanol (50:50, v/v). Its flow rate was 1.5 ml/min. The elution time for lycopene and ethyl-beta-apo-8'-carotenoate was approximately 11 and 5 min, respectively. Calibration curves of lycopene were linear in the concentration range of 3-200 ng/ml in plasma. Limits of detection and quantification in plasma were 1 and 4 ng/ml, respectively. Recovery was greater than 97%. Intra- and inter-day relative standard deviation for lycopene in plasma was less than 1.8 and 3.1%, respectively. This method was applied to the determination of lycopene plasma levels after single dose administration to dogs.
建立了一种在472 nm波长处检测的等度高效液相色谱法，用于犬血浆中番茄红素的测定，并对该方法进行了优化和验证。以乙基-β-apo-8'-胡萝卜素酸酯作为内标。使用粒径为5微米的Hypersil BDS RP-C18柱（150 mm×4.6 mm），用乙腈和甲醇（50:50，v/v）组成的流动相进行平衡。流速为1.5 ml/min。番茄红素和乙基-β-apo-8'-胡萝卜素酸酯的洗脱时间分别约为11分钟和5分钟。血浆中番茄红素的校准曲线在3-200 ng/ml浓度范围内呈线性。血浆中的检测限和定量限分别为1 ng/ml和4 ng/ml。回收率大于97%。血浆中番茄红素的日内和日间相对标准偏差分别小于1.8%和3.1%。该方法应用于犬单次给药后血浆中番茄红素水平的测定。
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