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基于高效液相色谱法的龈沟液中丙二醛定量分析。

Quantitation of malondialdehyde in gingival crevicular fluid by a high-performance liquid chromatography-based method.

机构信息

Department of Diagnosis and Surgery, Araraquara School of Dentistry, Universidade Estadual Paulista, Araraquara, São Paulo 14801-903, Brazil.

出版信息

Anal Biochem. 2012 Apr 1;423(1):141-6. doi: 10.1016/j.ab.2012.01.016. Epub 2012 Jan 28.

Abstract

Lipid peroxidation (LPO) has been associated with periodontal disease, and the evaluation of malondialdehyde (MDA) in the gingival crevicular fluid (GCF), an inflammatory exudate from the surrounding tissue of the periodontium, may be useful to clarify the role of LPO in the pathogenesis of periodontal disease. We describe the validation of a method to measure MDA in the GCF using high-performance liquid chromatography. MDA calibration curves were prepared with phosphate-buffered solution spiked with increasing known concentrations of MDA. Healthy and diseased GCF samples were collected from the same patient to avoid interindividual variability. MDA response was linear in the range measured, and excellent agreement was observed between added and detected concentrations of MDA. Samples' intra- and interday coefficients of variation were below 6.3% and 12.4%, respectively. The limit of quantitation (signal/noise=5) was 0.03 μM. When the validated method was applied to the GCF, excellent agreement was observed in the MDA quantitation from healthy and diseased sites, and diseased sites presented more MDA than healthy sites (P<0.05). In this study, a validated method for MDA quantitation in GCF was established with satisfactory sensitivity, precision, and accuracy.

摘要

脂质过氧化(LPO)与牙周病有关,评估龈沟液(GCF)中的丙二醛(MDA),GCF 是牙周组织周围组织的炎症渗出物,可能有助于阐明 LPO 在牙周病发病机制中的作用。我们描述了使用高效液相色谱法测量 GCF 中 MDA 的方法验证。用磷酸盐缓冲液制备 MDA 浓度逐渐增加的 MDA 校准曲线。从同一位患者中收集健康和患病的 GCF 样本,以避免个体间变异性。MDA 响应在测量范围内呈线性,并且观察到添加和检测到的 MDA 浓度之间具有极好的一致性。样品的日内和日间变异系数分别低于 6.3%和 12.4%。定量下限(信号/噪声=5)为 0.03 μM。当将经过验证的方法应用于 GCF 时,在健康和患病部位的 MDA 定量中观察到极好的一致性,并且患病部位的 MDA 含量高于健康部位(P<0.05)。在这项研究中,建立了一种用于 GCF 中 MDA 定量的经过验证的方法,该方法具有令人满意的灵敏度、精密度和准确性。

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