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肌氨酸作为前列腺癌进展的标志物:一种通过固相微萃取-气相色谱-三重四极杆质谱法快速、简单地定量检测人尿液中肌氨酸的方法。

Sarcosine as a marker in prostate cancer progression: a rapid and simple method for its quantification in human urine by solid-phase microextraction-gas chromatography-triple quadrupole mass spectrometry.

机构信息

Dipartimento di Chimica, Università della Calabria, Arcavacata di Rende, CS, Italy.

出版信息

Anal Bioanal Chem. 2011 Jul;400(9):2903-12. doi: 10.1007/s00216-011-4960-0. Epub 2011 Apr 15.

DOI:10.1007/s00216-011-4960-0
PMID:21491110
Abstract

Sarcosine is an amino acid derivative of N-methylglycine and is involved in the amino acid metabolism and methylation processes that are enriched during prostate cancer progression. It could also serve as a new target to be measured during therapeutic interventions and help in the identification of aggressive tumors for radical treatment. In this study, we present a new urine test that can help early diagnosis of prostate cancer. The method for the quantification of sarcosine in urine consists of a solid-phase microextraction (SPME) step followed by gas chromatography-triple quadrupole mass spectrometry analysis. We used a preliminary derivatization step with ethyl chloroformate/ethanol and the corresponding ester was then extracted by SPME in immersion mode. Several fibers were evaluated and the optimization of the parameters affecting the SPME process was carried out using an experimental design. The optimal values were 20 min extraction time, 10% NaCl, and 270°C using a divinylbenzene/Carboxen/polydimethylsiloxane fiber. The triple quadrupole analyzer acquired data in selected reaction monitoring mode, allowing us to obtain reconstructed chromatograms with well-defined chromatographic peaks. The accuracy and precision of this method were evaluated at concentrations of 70, 250, and 800 ng/ml and were found to be acceptable. Very satisfactory values (0.10 and 0.16 ng/ml, respectively) were also achieved for the limit of detection and the limit of quantification. The proposed protocol represents a rapid, simple, selective, and sensitive tool to quantify sarcosine in urine samples for prostate cancer diagnosis and for a screening test.

摘要

肌氨酸是 N-甲基甘氨酸的氨基酸衍生物,参与氨基酸代谢和甲基化过程,这些过程在前列腺癌进展过程中得到丰富。它也可以作为治疗干预期间测量的新靶标,并有助于识别需要根治性治疗的侵袭性肿瘤。在这项研究中,我们提出了一种新的尿液检测方法,可以帮助早期诊断前列腺癌。尿肌氨酸的定量方法包括固相微萃取 (SPME) 步骤,随后进行气相色谱-三重四极杆质谱分析。我们使用了初步的氯甲酸乙酯/乙醇衍生化步骤,然后通过 SPME 在沉浸模式下提取相应的酯。评估了几种纤维,并使用实验设计对影响 SPME 过程的参数进行了优化。最佳值为 20 分钟提取时间、10%NaCl 和 270°C,使用二乙烯基苯/Carboxen/聚二甲基硅氧烷纤维。三重四极杆分析仪以选择反应监测模式采集数据,使我们能够获得具有明确定义色谱峰的重构色谱图。该方法在 70、250 和 800ng/ml 浓度下的准确性和精密度均被评估为可接受。对于检测限和定量限,也获得了非常令人满意的值(分别为 0.10 和 0.16ng/ml)。所提出的方案代表了一种快速、简单、选择性和灵敏的工具,可用于定量尿液样本中的肌氨酸,用于前列腺癌诊断和筛查测试。

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