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Introduction

Material & Methods

LC-MS/MS

Injection volume of 1–20 µl is used. Chromatographic separation is performed on an Analytical Column (order no. 63100) at a column temperature of 40 °C. The analytes are separated under isocratic conditions at a flow rate of 0.4 ml/min using Mobile Phase (order no. 63001).

Electrospray ionization in negative ion mode is used. The multiple reaction monitoring (MRM) transitions used for quantification are listed in Table 1.

A full calibration of the analysis system for each series of measurements is performed with 6PLUS1® calibrator (order no. 63039). Calibration curves are constructed by calculating the analyte to internal standard (ISTD) peak area ratio on the y axis against calibrator concentrations on the x axis. Then a calibration curve is plotted for both PEth homolouges using linear regression and 1/x weighting.

Table 1

Results

The run time is 3.5 min (see chromatogram in fig. 1). The calibration range of 10-1000 ng/mL covers the forensic and clinical relevant concentrations (table 2).

Table 2 

Conclusion

This application note describes a protocol for the determination of PEth 16:0/18:1 and PEth 16:0/18:2 in whole blood using Chromsystems components. 

Fig. 1

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RUO: For research use only and not for diagnostic purposes