毛細管電泳法對透析病患血清中硫酸吲哚酚及硫酸對甲酚之分析研究

本研究建立一簡單、靈敏及快速的毛細管電泳分析法，利用線上濃縮分離技術－電場放大樣品堆積及微胞掃集電動層析法(FASS-sweeping-MEKC)，同時分析人體血清中兩種內生性親蛋白質尿毒素: 硫酸對甲酚(para-cresol sulfate, PCS)及硫酸吲哚酚(3-indoxyl sulfate, 3-INS)。以慢性腎衰竭病患而言，由於腎功能的衰退，PCS與3-INS會大量的累積在體內，進而造成許多負面的影響和毒性，在臨床上已漸受重視，故其檢測方法的建立刻不容緩。本法利用含150 mM十二基氫硫酸鈉鹽及10%甲醇之磷酸鹽溶液(100 mM, pH 4.0)作為分析用的緩衝溶液，搭配214 nm的偵測波長，10分鐘內即可完成分離，進行定性與定量分析。血清中PCS與3-INS的偵測極限分別為100及30 ng/mL (S/N=3)。本法之確效結果顯示各待測物在定量範圍內均具有良好的線性關係(r≧0.99)；而在同日內及異日間的精密度與準確度探討，相對標準偏差(RSD)與相對誤差(RE)皆小於3.47%。此外，本研究方法已實際應用於洗腎病患體內尿毒素的分析，期盼在臨床上可協助評估病患疾病嚴重程度及預後之情形，同時也輔助作為投予藥物治療時的客觀性指標。

關鍵字

硫酸吲哚酚；硫酸對甲酚

並列摘要

An efficient, simple, sensitive and rapid on-line sample concentration technique－field-amplified sample stacking-sweeping-micellar electrokinetic chromatography (FASS-sweeping-MEKC) was developed for simultaneous determination of para-cresol sulfate (PCS) and 3-indoxyl sulfate (3-INS) in human serum. PCS and 3-INS are the two important representatives of endogenous “protein-bound uremic toxins” and would accumulate in chronic renal failure patients with impaired kidney functions, leading to various adverse impacts and toxicities on human body. Therefore, detection of PCS and 3-INS are getting more and more attention in recent years at clinic. In this study, qualitative and quantitative analysis of PCS and 3-INS in human serum can be achieved under the optimized conditions. Separation was carried out in phosphate buffer (100 mM, pH 4.0) containing 150 mM SDS and 10% MeOH at 214 nm within 10 min. Detection limit (S/N=3) was found to be 100 ng/mL for PCS and 30 ng/mL for 3-INS. Calibration curves showed great linearity (r≧0.99) and small RSD and RE values (≦3.47%) in intra-day and inter-day assays demonstrated the good precision and accuracy of the CE method. Furthermore, the developed method has been successfully applied to analyze real samples from hemodialysis patients, aiming to be helpful for disease evaluation and become as an objective indicator for medication treatment in patients.

並列關鍵字

3-indoxyl sulfate ； para-cresol sulfate

參考文獻

[1] National Health Insurance Administration, Ministry of Health and Welfare中華民國衛生福利部中央健康保險署－慢性腎臟病防治(2014)

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[2] National Kidney Foundation-Kidney Disease Outcomes Quality Initiative, KDOQI clinical practice guideline for diabetes and chronic kidney disease (2012)