Development of a method to measure free and bound ropivacaine in human plasma using equilibrium dialysis and Hydrophilic interaction chromatography coupled to high resolution mass spectrometry

M. Abbas, L. Ahmad, Y. Shah, M. Gill, D.G. Watson

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12 Citations (Scopus)

Abstract

The pharmacodynamics of absorption of local anaesthetics used during surgical procedures into tissues is governed by the amount of free drug in plasma. Toxicity may occur during continuous infusions if the levels of free drug become too high which may occur if the binding capacity of the α-1- acid glycoprotein present in plasma is exceeded. In order to monitor this a method was developed for the determination of the amount of free and bound ropivacaine in human plasma during knee and hip surgery. Rapid equilibrium dialysis units were used to separate free and bound drug then protein and buffer salts were removed by solvent precipitation. Analysis was carried out using a ZICHILIC HPLC column interfaced with an LTQ Orbitrap mass spectrometer. The following extracted ion ranges ([M+H]) were monitored: m/z 275.21-275.22 for ropivacaine and m/z 235.175-235.185 for lidocaine. The method was calibrated by spiking ropivacaine, and a fixed amount of lidocaine as internal standard, into plasma over the range 0.01-1.5 μg/ml. The equation of the line was y=0.886x±4.2% (n=2), forcing the curve through zero since blank plasma was free of the analyte. The values obtained for the accuracy and precision of the analysis of plasma spiked at 0.03 μg/ml and 1.5 μg/ml were 93.2%±2.8% and 95.4%±1.5% respectively (n=5). Repeat analysis of a patient sample for free and bound drug gave the following values for levels of ropivacaine: bound 1.63 μg/ml±1.48%, unbound 0.0671 μg/ml±1.68% (n=5).
Original languageEnglish
Pages (from-to)60-63
Number of pages4
JournalTalanta
Volume117
DOIs
Publication statusPublished - 15 Dec 2013

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Plasma (human)
Dialysis
Chromatography
Hydrophobic and Hydrophilic Interactions
Mass spectrometry
Mass Spectrometry
Plasmas
Lidocaine
Pharmaceutical Preparations
Pharmacodynamics
Mass spectrometers
Local Anesthetics
Surgery
Toxicity
Glycoproteins
Buffers
Salts
ropivacaine
Hip
Ions

Keywords

  • method
  • free ropivacaine
  • measure
  • bound ropivacaine
  • high resolution
  • mass spectrometry
  • human plasma
  • equilibrium dialysis
  • hydrophilic interaction
  • chromatography

Cite this

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title = "Development of a method to measure free and bound ropivacaine in human plasma using equilibrium dialysis and Hydrophilic interaction chromatography coupled to high resolution mass spectrometry",
abstract = "The pharmacodynamics of absorption of local anaesthetics used during surgical procedures into tissues is governed by the amount of free drug in plasma. Toxicity may occur during continuous infusions if the levels of free drug become too high which may occur if the binding capacity of the α-1- acid glycoprotein present in plasma is exceeded. In order to monitor this a method was developed for the determination of the amount of free and bound ropivacaine in human plasma during knee and hip surgery. Rapid equilibrium dialysis units were used to separate free and bound drug then protein and buffer salts were removed by solvent precipitation. Analysis was carried out using a ZICHILIC HPLC column interfaced with an LTQ Orbitrap mass spectrometer. The following extracted ion ranges ([M+H]) were monitored: m/z 275.21-275.22 for ropivacaine and m/z 235.175-235.185 for lidocaine. The method was calibrated by spiking ropivacaine, and a fixed amount of lidocaine as internal standard, into plasma over the range 0.01-1.5 μg/ml. The equation of the line was y=0.886x±4.2{\%} (n=2), forcing the curve through zero since blank plasma was free of the analyte. The values obtained for the accuracy and precision of the analysis of plasma spiked at 0.03 μg/ml and 1.5 μg/ml were 93.2{\%}±2.8{\%} and 95.4{\%}±1.5{\%} respectively (n=5). Repeat analysis of a patient sample for free and bound drug gave the following values for levels of ropivacaine: bound 1.63 μg/ml±1.48{\%}, unbound 0.0671 μg/ml±1.68{\%} (n=5).",
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author = "M. Abbas and L. Ahmad and Y. Shah and M. Gill and D.G. Watson",
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T1 - Development of a method to measure free and bound ropivacaine in human plasma using equilibrium dialysis and Hydrophilic interaction chromatography coupled to high resolution mass spectrometry

AU - Abbas, M.

AU - Ahmad, L.

AU - Shah, Y.

AU - Gill, M.

AU - Watson, D.G.

PY - 2013/12/15

Y1 - 2013/12/15

N2 - The pharmacodynamics of absorption of local anaesthetics used during surgical procedures into tissues is governed by the amount of free drug in plasma. Toxicity may occur during continuous infusions if the levels of free drug become too high which may occur if the binding capacity of the α-1- acid glycoprotein present in plasma is exceeded. In order to monitor this a method was developed for the determination of the amount of free and bound ropivacaine in human plasma during knee and hip surgery. Rapid equilibrium dialysis units were used to separate free and bound drug then protein and buffer salts were removed by solvent precipitation. Analysis was carried out using a ZICHILIC HPLC column interfaced with an LTQ Orbitrap mass spectrometer. The following extracted ion ranges ([M+H]) were monitored: m/z 275.21-275.22 for ropivacaine and m/z 235.175-235.185 for lidocaine. The method was calibrated by spiking ropivacaine, and a fixed amount of lidocaine as internal standard, into plasma over the range 0.01-1.5 μg/ml. The equation of the line was y=0.886x±4.2% (n=2), forcing the curve through zero since blank plasma was free of the analyte. The values obtained for the accuracy and precision of the analysis of plasma spiked at 0.03 μg/ml and 1.5 μg/ml were 93.2%±2.8% and 95.4%±1.5% respectively (n=5). Repeat analysis of a patient sample for free and bound drug gave the following values for levels of ropivacaine: bound 1.63 μg/ml±1.48%, unbound 0.0671 μg/ml±1.68% (n=5).

AB - The pharmacodynamics of absorption of local anaesthetics used during surgical procedures into tissues is governed by the amount of free drug in plasma. Toxicity may occur during continuous infusions if the levels of free drug become too high which may occur if the binding capacity of the α-1- acid glycoprotein present in plasma is exceeded. In order to monitor this a method was developed for the determination of the amount of free and bound ropivacaine in human plasma during knee and hip surgery. Rapid equilibrium dialysis units were used to separate free and bound drug then protein and buffer salts were removed by solvent precipitation. Analysis was carried out using a ZICHILIC HPLC column interfaced with an LTQ Orbitrap mass spectrometer. The following extracted ion ranges ([M+H]) were monitored: m/z 275.21-275.22 for ropivacaine and m/z 235.175-235.185 for lidocaine. The method was calibrated by spiking ropivacaine, and a fixed amount of lidocaine as internal standard, into plasma over the range 0.01-1.5 μg/ml. The equation of the line was y=0.886x±4.2% (n=2), forcing the curve through zero since blank plasma was free of the analyte. The values obtained for the accuracy and precision of the analysis of plasma spiked at 0.03 μg/ml and 1.5 μg/ml were 93.2%±2.8% and 95.4%±1.5% respectively (n=5). Repeat analysis of a patient sample for free and bound drug gave the following values for levels of ropivacaine: bound 1.63 μg/ml±1.48%, unbound 0.0671 μg/ml±1.68% (n=5).

KW - method

KW - free ropivacaine

KW - measure

KW - bound ropivacaine

KW - high resolution

KW - mass spectrometry

KW - human plasma

KW - equilibrium dialysis

KW - hydrophilic interaction

KW - chromatography

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U2 - 10.1016/j.talanta.2013.08.049

DO - 10.1016/j.talanta.2013.08.049

M3 - Article

VL - 117

SP - 60

EP - 63

JO - Talanta

JF - Talanta

SN - 0039-9140

ER -