Robinson-Fuentes V A, Jaime-Sánchez J L, García-Aguilar L, Gómez-Peralta M, Vázquez-Garcidueñas M S, Vázquez-Marrufo G
División de Estudios de Posgrado, Facultad de Ciencias Médicas y Biológicas Dr. Ignacio Chávez, Universidad Michoacana de San Nicolás de Hidalgo, Morelia, Michoacán, Mexico.
J Pharm Biomed Anal. 2008 Aug 5;47(4-5):913-7. doi: 10.1016/j.jpba.2008.03.032. Epub 2008 Apr 10.
Amanitins are toxins found in species of the mushroom genera Amanita, Lepiota and Galerina. Intoxication after ingestion of these mushrooms can be fatal with an estimated 20% of mortality rate. An early diagnosis is necessary in order to avoid invasive and expensive therapy and to improve patient's prognosis. In this paper, a Capillary Zone Electrophoresis method was developed and validated to determine alpha- and beta-amanitin in urine in less than 7 min using 5 mM, pH 10 borate buffer as background electrolyte. The separation conditions were: capillary: 75 microm I.D., 41 cm effective length, 48 cm total length, 25 degrees C, 20 KV and PDA detection at 214 nm. Sample treatment for analysis only required urine dilution in background electrolyte. The method was validated following established criteria and was found to be selective, linear in the range 5-100 ng/ml. Intra- and inter-day precision and accuracy were within required limits. Limit of detection (LOD) and limit of quantification (LOQ) were 1.5 and 5 ng/ml, respectively. Eight urine samples from suspected cases of intoxication with amanitins were analyzed after 2 years of storage at -20 degrees C, and beta-amanitin was determined in two samples with concentrations of 53 and 65 ng/ml, respectively. The method here described includes the use of non-aggressive reagents to the capillary or the system and is the first Capillary Electrophoresis method used to determine amanitins in clinical samples.
鹅膏毒素是在鹅膏菌属、环柄菇属和盔孢伞属蘑菇中发现的毒素。摄入这些蘑菇后的中毒可能是致命的，估计死亡率为20%。为了避免侵入性和昂贵的治疗并改善患者的预后，早期诊断是必要的。在本文中，开发并验证了一种毛细管区带电泳方法，该方法使用5 mM、pH值为10的硼酸盐缓冲液作为背景电解质，可在不到7分钟的时间内测定尿液中的α-和β-鹅膏毒素。分离条件为：毛细管：内径75微米，有效长度41厘米，总长度48厘米，25℃，20 kV，在214纳米处进行光电二极管阵列检测。分析所需的样品处理仅需将尿液稀释在背景电解质中。该方法按照既定标准进行了验证，结果表明具有选择性，在5-100 ng/ml范围内呈线性。日内和日间精密度及准确度均在规定范围内。检测限（LOD）和定量限（LOQ）分别为1.5和5 ng/ml。对来自疑似鹅膏毒素中毒病例的8份尿液样本在-20℃下储存2年后进行了分析，在两份样本中分别测定出β-鹅膏毒素的浓度为53和65 ng/ml。本文所述方法使用的试剂对毛细管或系统无侵蚀性，是用于测定临床样本中鹅膏毒素的首个毛细管电泳方法。
