Division of Organic Chemistry and Biochemistry, Ruđer Bošković Institute, Zagreb, Croatia.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Dec 15;879(32):3920-6. doi: 10.1016/j.jchromb.2011.11.003. Epub 2011 Nov 9.
Opiorphin, QRFSR-peptide, is a mature product of the PROL1 (proline rich, lacrimal 1) protein that showed beneficial effects in pain management, antidepressant-like actions as well as involvement in colonic motility and erectile physiology. Using opiorphin as a potential biomarker of different pathological states requires the development of robust and sensitive methods. We report a highly sensitive and specific liquid chromatography with tandem mass spectrometric detection (LC-MS/MS) analytical method for the analysis of opiorphin in human saliva. Quantification was based on multiple reaction monitoring using characteristic transitions (m/z 347/120 - as quantifying ion; 347/175 and 347/268 as qualifying ions). The assay was linear in the range of 0-110 ng/ml and the lower limit of quantification reached was 1.0 ng/ml. The intra-day precision and accuracy were between 2.7-5.6% and -2.3 to 3.2%, respectively. The inter-day precision and accuracy were between 10.8-13.7% and -11.0 to 52%, respectively. Mean recovery was 106% and mean matrix effect was 0.97. Opiorphin in TFA treated saliva samples was stable for at least 12h at room temperature and up to 30 days at -20°C. Opiorphin levels in human saliva samples collected from young healthy individuals ranged from 2.8 to 25.9 ng/ml.
阿片啡肽（Opiorphin）是富含脯氨酸的泪液蛋白 1（PROL1）蛋白的成熟产物，在疼痛管理、抗抑郁作用以及参与结肠动力和勃起生理方面具有有益的效果。将阿片啡肽用作不同病理状态的潜在生物标志物需要开发强大而敏感的方法。我们报告了一种用于分析人唾液中阿片啡肽的高灵敏度和特异性的液相色谱-串联质谱检测（LC-MS/MS）分析方法。定量基于使用特征过渡（m/z 347/120-作为定量离子；347/175 和 347/268 作为定性离子）的多重反应监测。该测定在 0-110ng/ml 范围内呈线性，达到的定量下限为 1.0ng/ml。日内精密度和准确度分别在 2.7-5.6%和-2.3 至 3.2%之间。日间精密度和准确度分别在 10.8-13.7%和-11.0 至 52%之间。平均回收率为 106%，平均基质效应为 0.97。TFA 处理的唾液样品中的阿片啡肽在室温下至少稳定 12 小时，在-20°C 下稳定长达 30 天。从年轻健康个体收集的人唾液样本中的阿片啡肽水平范围为 2.8 至 25.9ng/ml。
