![Reverse-phase high performance liquid chromatography for the determination of tiopronin in human plasma after derivatization with p-bromophenacyl bromide [An article from: Analytica Chimica Acta]](https://www.ebooknetworking.net/books/B00/0RR/bigB000RR8JNQ.jpg)
Reverse-phase high performance liquid chromatography for the determination of tiopronin in human plasma after derivatization with p-bromophenacyl bromide [An article from: Analytica Chimica Acta]
Description
This digital document is a journal article from Analytica Chimica Acta, published by Elsevier in 2006. The article is delivered in HTML format and is available in your Amazon.com Media Library immediately after purchase. You can view it with any web browser.
Description:
In the study, we developed a simple, rapid and sensitive method for the determination of tiopronin (TP) in human plasma, which was based on derivatization with p-bromophenacyl bromide (p-BPB) followed by liquid-liquid extraction and reverse-phase HPLC-UV detection. For the first time, the p-BPB was introduced into the derivatization of TP. The thiol group of TP was trapped with p-BPB to form a TP-p-BPB adduct, which can be very suitable for UV detection. From acidified plasma samples, the derivatized TP was extracted with 5mL dichloromethane. Effective chromatographic separation was achieved using a C18 column (DIAMONSIL 150mmx4mm i.d., 5@mm) based on an acetonitrile-water-trifluoroacetic acid (40:59.88:0.12, v/v/v) elution at a flow-rate of 1mL/min. The IS and the derivatized TP were detected at 263nm. No endogenous substances were found to interfere. The limit of quantification for derivatized TP (TP-p-BPB) in plasma was 40ng/mL. The calibration curve for the derivatized TP showed linearity in the range 0.04-4@mg/mL with a regression coefficient corresponding to 0.9991 and the coefficient of the variation of the points of the calibration curve being lower than 10%. Extraction recoveries of the derivatized TP in plasma were greater than 72%. The method was suitably validated and successfully applied to determination of TP in human plasma samples.
Description:
In the study, we developed a simple, rapid and sensitive method for the determination of tiopronin (TP) in human plasma, which was based on derivatization with p-bromophenacyl bromide (p-BPB) followed by liquid-liquid extraction and reverse-phase HPLC-UV detection. For the first time, the p-BPB was introduced into the derivatization of TP. The thiol group of TP was trapped with p-BPB to form a TP-p-BPB adduct, which can be very suitable for UV detection. From acidified plasma samples, the derivatized TP was extracted with 5mL dichloromethane. Effective chromatographic separation was achieved using a C18 column (DIAMONSIL 150mmx4mm i.d., 5@mm) based on an acetonitrile-water-trifluoroacetic acid (40:59.88:0.12, v/v/v) elution at a flow-rate of 1mL/min. The IS and the derivatized TP were detected at 263nm. No endogenous substances were found to interfere. The limit of quantification for derivatized TP (TP-p-BPB) in plasma was 40ng/mL. The calibration curve for the derivatized TP showed linearity in the range 0.04-4@mg/mL with a regression coefficient corresponding to 0.9991 and the coefficient of the variation of the points of the calibration curve being lower than 10%. Extraction recoveries of the derivatized TP in plasma were greater than 72%. The method was suitably validated and successfully applied to determination of TP in human plasma samples.
