The Application Notebook
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.
Click here to view full-size graphic
Click here to view full-size graphic
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 www.mn-net.com/apps
(2) Application No. 128200, MACHEREY-NAGEL, available from www.mn-net.com/apps
Macherey-Nagel GmbH & Co. KG
Neumann-Neander-Str. 6–8, 52355 Düren, Germany
tel. +49-(0)2421-969-0
Website: www.mn-net.com
Inside the Laboratory: The Gionfriddo Group at the University at Buffalo
March 28th 2024In this edition of “Inside the Laboratory,” Emanuela Gionfriddo, PhD, an associate professor of chemistry at the University at Buffalo, discusses her group’s current research endeavors, including using solid-phase microextraction (SPME) coupled to liquid chromatography (LC) and gas chromatography (GC) to further understand the chemical relationship between environmental exposure and disease and elucidate micropollutants fate in the environment and biological systems.
Transferring Methods to Compact and Portable HPLC
February 14th 2024The current trend in laboratory equipment design is the miniaturization of laboratory instruments. Smaller-scale HPLC instruments offer benefits that cannot be matched by analytical-scale equipment, especially in the areas of portability, reduced fluid volumes, and reduced operating costs. Yet, the miniaturization of laboratory equipment has brought with it a unique set of challenges, including transferring methods to compact LC. Capillary LC expands the use of LC to applications not currently done using conventional LC in a wide array of application areas, including pharmaceutical, food and beverage, petrochemical, environmental, and oil and gas. Greg Ward, Axcend’s CEO wrote, “Customers want an HPLC system with a small footprint, low flow rates and green chemistry.” Join his podcast where he shares method transfer in these application areas.