Determination of Pharmaceuticals from Serum

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The Application Notebook

The Application Notebook, The Application Notebook-09-02-2018, Volume 36, Issue 9
Pages: 716–717

This application note describes the determination of pharmaceuticals from serum using solid-phase extraction with the hydrophilic-lipophilic balanced SPE phase CHROMABOND® HLB for analyte enrichment and for sample clean-up. The eluates from SPE are finally analyzed by HPLC–MS/MS on a NUCLEOSHELL® PFP core-shell phase.

Nowadays, people are suffering from various diseases. Therefore, they are prescribed many types of pharmaceuticals as part of their treatment, for instance anesthetics, antibiotics, anticholinergics, anticonvulsants, etc. In order for the treatment to be successful, it is necessary to keep controlling the levels of the pharmaceuticals to provide an accurate dosage. This leads to an increasing demand for the development of accurate and sensitive analytical methods for the pharmaceuticals from serum to protect human health.

Table I: SRM transitions for the investigated pharmaceuticals.

Solid-Phase Extraction (1)

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Subsequent Analysis: HPLC–MS/MS (2)


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The recovery rates show that the determination of pharmaceuticals from serum could be carried out successfully (Figure 2). By using SPE with CHROMABOND HLB it was possible to recover nearly all pharmaceuticals from serum on average with good reproducibility. Regarding the different types pharmaceuticals of the average recovery rates were: for anesthetics 90.8%, for antibiotics 94.4%, for anticholinergics 84.8%, for anticonvulsants 97.7%, for antidepressants 77.4%, for antihistamines 87.1%, for anti-inflammatory drugs 84.1%, for beta blockers 89.5%, for calcium channel blockers 107.5%, and for diuretics 87.7%

Figure 2: Recovery rates for solid-phase extraction method of pharmaceuticals from serum.

The identification and quantification of pharmaceuticals in the solid-phase extracts were carried out by ESI mass spectrometry on an EC 50/2 NUCLEOSHELL PFP column. The chromatogram in Figure 1 shows the results of solid-phase eluate spiked with 10 ng/mL serum for each pharmaceutical.

Figure 1: Chromatogram of serum sample spiked with 10 ng/mL for each pharmaceutical.


The presented application describes a quick and convenient method for the determination of pharmaceuticals from serum by SPE with a hydrophilic-lipophilic balanced phase, followed by HPLC–MS/MS analysis.


(1) Application No. 306510, MACHEREY-NAGEL, available from

(2) Application No. 128200, MACHEREY-NAGEL, available from

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