Moradi Milad, Soleymani Jafar, Tayebi-Khosroshahi Hamid, Khoubnasabjafari Maryam, Jouyban Abolghasem
Pharmaceutical Analysis Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Liver and Gastrointestinal Diseases Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Iran J Pharm Res. 2021 Spring;20(2):68-78. doi: 10.22037/ijpr.2020.114330.14799.
The development of simple, fast, cheap and reliable analytical methods for tracing biological indicators is demanded through clinical investigations. Herein, we developed, for the first time, a cheap and specific method for the extraction and quantification of p-cresol (pC) in real plasma samples of chronic kidney disease (CKD). Plasma samples were prepared by hydrolyzing in an acidic medium to convert pCS (p-cresol sulfate) and p-Cresol glucuronide (pCG) to pC. Next, proteins of plasma samples were precipitated and then pC was extracted by acetonitrile (ACN) and saturated NaCl (as salting-out agent). Finally, fluorescence emissions were measured at λ/λ = 280/310 nm. The specificity of the method was checked by testing various possible interfering agents. The obtained results revealed a specific determination of pC. Under optimal conditions, a linear range was detected from 0.5 to 30 µg/mL of pC with a lower limit of detection (LLOQ) of 0.5 µg/mL. The reliability of the method was checked by calculating the repeatability, selectivity, and accuracy of the developed method for pC determination in plasma samples. The application of the developed method was investigated for the detection of pC in a number of CKD patients. Due to the simplicity and selectivity, the developed method could be applied for routine analysis of pC concentrations in the plasma samples of CKD patients. In addition, the developed method showed great potential for developing a point-of-care testing (POCT) device.
临床研究需要开发简单、快速、廉价且可靠的生物指标追踪分析方法。在此,我们首次开发了一种廉价且特异的方法,用于慢性肾脏病(CKD)患者实际血浆样本中对甲酚(pC)的提取和定量。血浆样本通过在酸性介质中水解,将对甲酚硫酸盐(pCS)和对甲酚葡萄糖醛酸苷(pCG)转化为pC来制备。接下来,沉淀血浆样本中的蛋白质,然后用乙腈(ACN)和饱和氯化钠(作为盐析剂)提取pC。最后,在λ/λ = 280/310 nm处测量荧光发射。通过测试各种可能的干扰剂来检验该方法的特异性。所得结果显示可特异性测定pC。在最佳条件下,检测到pC的线性范围为0.5至30 µg/mL,最低检测限(LLOQ)为0.5 µg/mL。通过计算所开发方法在血浆样本中测定pC的重复性、选择性和准确性来检验该方法的可靠性。研究了所开发方法在一些CKD患者中检测pC的应用。由于方法的简单性和选择性,所开发的方法可用于CKD患者血浆样本中pC浓度的常规分析。此外,所开发的方法在开发即时检测(POCT)设备方面显示出巨大潜力。