Antunes Alisson Henrique, Faria Flávia Rasmussen, Mota João Felipe, Santiago Mariângela Fontes, Kogawa Ana Carolina, Rezende Kênnia Rocha
Laboratório de Biofarmácia e Farmacocinética (BioPk), Faculdade de Farmácia, Universidade Federal de Goiás, Goiânia, Goiás, Brazil.
Laboratório de Investigação em Nutrição Clínica e Esportiva (LABINCE), Universidade Federal de Goiás, Goiânia, Goiás, Brazil.
Saudi Pharm J. 2020 May;28(5):599-606. doi: 10.1016/j.jsps.2020.03.012. Epub 2020 Apr 2.
In sports, curcumin, a substance derived from the rhizome of (turmeric) plant with antioxidant effect 8 times greater than vitamin E, has attracted the attention of scientists because of its potent antioxidant action, since in athletes subjected to intense exercise the-endogenous mechanisms of neutralization of reactive species are saturated. However, the pharmacokinetic characteristics of curcumin do not favor its medicinal use due to its low absorption, accelerated metabolism and rapid systemic elimination. Thus, the determination of plasma levels in supplemented patients is a crucial step in their pharmacodynamic evaluation. Therefore, the objective of this work was to develop and validate an analytical method by HPLC-FLD for curcumin evaluation in plasma of supplemented athletes. Luna column (C18; 150 × 4 mm; 3 µm), acetonitrile: acetic acid pH 3.2 (45:55 to 60:40) as mobile phase, flow rate of 1 mL min, excitation at 429/285 nm and emission at 529 nm and injection of 10 µL were the chromatographic conditions used. Plasma samples were extracted using ethylacetate and methanol (95: 5, 500 µL) and estradiol (30 µg mL) as internal standard, with subsequent stirring (3 min) and centrifugation (8 min) (triple extraction). The organic fraction was evaporated under N (20 min) and the dried residue reconstituted in acetonitrile. The method was linear between 44 and 261 ng mL, showing intra-day (2.05.6%) and inter-day (4.0-5.1%) precision with accuracy and selectiveness (curcumin t = 8.7 min and internal standard t = 13.9 min with relative recovery of 83.2%). So, it can be successfully used for curcumin evaluation in plasma samples from supplemented athletes, as well as being an alternative and advantageous method to UV-Vis and MS/MS in bioavailability studies.
在体育领域，姜黄素是一种从姜黄属植物根茎中提取的物质，其抗氧化作用比维生素E强8倍，由于其强大的抗氧化作用而引起了科学家们的关注，因为在进行高强度运动的运动员中，内源性活性物质中和机制会饱和。然而，姜黄素的药代动力学特性不利于其药用，因为它吸收低、代谢快且全身消除迅速。因此，测定补充姜黄素患者的血浆水平是其药效学评估的关键步骤。因此，本研究的目的是开发并验证一种采用高效液相色谱-荧光检测法（HPLC-FLD）测定补充姜黄素运动员血浆中姜黄素的分析方法。色谱条件如下：使用月旭柱（C18；150×4mm；3μm），以乙腈：pH3.2的乙酸（45:55至60:40）作为流动相，流速为1mL/min，激发波长为429/285nm，发射波长为529nm，进样量为10μL。血浆样品用乙酸乙酯和甲醇（95:5，500μL）提取，并以雌二醇（30μg/mL）作为内标，随后搅拌（3分钟）并离心（8分钟）（三次萃取）。有机相在氮气下蒸发（20分钟），干燥残渣用乙腈复溶。该方法在44至261ng/mL之间呈线性，日内精密度为2.0-5.6%，日间精密度为4.0-5.1%，具有准确性和选择性（姜黄素保留时间t=8.7分钟，内标保留时间t=13.9分钟，相对回收率为83.2%）。因此，该方法可成功用于测定补充姜黄素运动员血浆样品中的姜黄素，并且在生物利用度研究中是紫外可见分光光度法和串联质谱法的一种替代且具有优势的方法。
