Determination of Retinol, α‐Tocopherol, Lycopene, and β‐Carotene in Human Plasma Using HPLC with UV‐Vis Detection: Application to a Clinical Study

Abstract

A method is described here for the simultaneous determination of retinol,α-tocopherol, lycopene, andβ-carotene in human plasma. The effectiveness of various protein precipitants and extraction solvents was tested. After adequate sample preparation, the samples were injected directly into the HPLC system. The separation was realized on an analytical reversed-phase column with a UV-Vis detection. The analytical performance of this method was satisfactory. The intraassay and interassay coefficients of variation were below 10%. The recoveries were as follows: 97.0% (CV 2.4%) for retinol, 94.6% (CV 1.7%) forα-tocopherol, 91.9% (CV 3.6%) for lycopene, and 93.9% (CV 4.2%) forβ-carotene. The levels of selected fat-soluble vitamins in plasma of patients with cardiovascular disease were measured and discussed.

Personalized Cardio‐Metabolic Responses to an Anti‐Inflammatory Nutrition Intervention in Obese Adolescents: A Randomized Controlled Crossover Trial

“…Plasma glucose, total cholesterol, HDL cholesterol, TAG, and apolipoprotein A1 (ApoA1) were measured using RX Daytona, as described previously. [35] Plasma nonesterified fatty acid (NEFA) concentration was measured by an enzymatic assay kit (WAKO Diagnostics, USA). Intra-and inter-assay CVs were <10% for all analyses.…”

“…ELISA determined total and HMW adiponectin, soluble CD163 (sCD163), IL‐6, tumor necrosis factor‐α (TNF‐α; R&D Systems, UK), insulin (Mercodia, Sweden), complement component C3 (AssayPro, MO, USA), and fetuin‐A (BioVendor, Czech Republic). Plasma glucose, total cholesterol, HDL cholesterol, TAG, and apolipoprotein A1 (ApoA1) were measured using RX Daytona, as described previously . Plasma nonesterified fatty acid (NEFA) concentration was measured by an enzymatic assay kit (WAKO Diagnostics, USA).…”

Liquid chromatography in clinical laboratory diagnostics

“…Была разработана и внедрена удобная и быстрая методика для количественно определения бета-каротина в сыворотке/плазме крови. Пробо-подготовка включала в себя этап осаждения белков и экстракции, причем полученный в результате это-го экстракт готов для ввода в хроматографическую изократическую систему со спектрофотометриче-ским детектором [16]. Хроматографирование проис-ходит на обращенно-фазной колонке, что позволяет разделить в одной пробе несколько форм каротино-идов: альфа-, цис-бета-и транс-бета-каротин; при этом время анализа не превышает 10 мин.…”

Rapid determination of lipophilic vitamins in human serum by ultra‐high performance liquid chromatography using a fluorinated column and high‐throughput miniaturized liquid–liquid extraction

“…S1. Numerous LC methods have been published regarding the simultaneous determination of retinol and selected tocopherol isomers [7][8][9][10][11][12]. Most frequently, these methods are carried out by HPLC and UHPLC.…”

“…Tocopherol isomers and retinol can be determined in a broad spectrum of biological materials, such as plasma, serum, seminal plasma, red blood cells, and human milk [7][8][9][10][11][12]15]. The most common sample preparation procedure for their subsequent determination is protein precipitation followed by liquid-liquid extraction (LLE).…”