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Determination of Vitamin C (Ascorbic Acid) Using High Performance Liquid Chromatography Coupled with Electrochemical Detection

机译:高效液相色谱-电化学检测法测定维生素C(抗坏血酸)

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Vitamin C (ascorbic acid, ascorbate, AA) is a water soluble organic compound that participates in many biological processes. The main aim of this paper was to utilize two electrochemical detectors (amperometric – Coulouchem III and coulometric – CoulArray) coupled with flow injection analysis for the detection of ascorbic acid. Primarily, we optimized the experimental conditions. The optimized conditions were as follows: detector potential 100 mV, temperature 25 °C, mobile phase 0.09% TFA:ACN, 3:97 (v/v) and flow rate 0.13 mL·min-1. The tangents of the calibration curves were 0.3788 for the coulometric method and 0.0136 for the amperometric one. The tangent of the calibration curve measured by the coulometric detector was almost 30 times higher than the tangent measured by the amperometric detector. Consequently, we coupled a CoulArray electrochemical detector with high performance liquid chromatography and estimated the detection limit for AA as 90 nM (450 fmol per 5 μL injection). The method was used for the determination of vitamin C in a pharmaceutical preparations (98 ± 2 mg per tablet), in oranges (Citrus aurantium) (varied from 30 to 56 mg/100 g fresh weight), in apples (Malus sp.) (varied from 11 to 19 mg/100 g fresh weight), and in human blood serum (varied from 38 to 78 μM). The recoveries were also determined.
机译:维生素C(抗坏血酸,抗坏血酸,AA)是一种水溶性有机化合物,参与许多生物过程。本文的主要目的是利用两个电化学检测器(安培– Coulouchem III和库仑– CoulArray)结合流动注射分析法检测抗坏血酸。首先,我们优化了实验条件。优化的条件如下:检测器电位100 mV,温度25°C,流动相0.09%TFA:ACN,3:97(v / v),流速0.13 mL·min -1 。电量法的校准曲线的切线为0.3788,电流法的校准曲线的切线为0.0136。库仑检测器测得的校准曲线的切线几乎比安培检测器测得的切线高30倍。因此,我们将CoulArray电化学检测器与高效液相色谱仪结合使用,估计AA的检测限为90 nM(每5μL进样450 fmol)。该方法用于测定药物制剂中的维生素C(每片98±2 mg),桔子(柑桔)(鲜重30至56 mg / 100 g)和苹果(Malus sp。)中的维生素C。 (在11至19 mg / 100 g新鲜体重之间变化)和人血清中(在38至78μM之间变化)。还确定了回收率。

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