Ultra‐performance liquid chromatography coupled to linear ion trap mass spectrometry for the identification of drug metabolites in biological samples

Abstract

The coupling of ultra-performance liquid chromatography, operating at elevated pressures, to a linear ion trap mass spectrometer provides a high-performance system suitable for drug metabolite characterisation. This system demonstrates improved chromatographic efficiency and sensitivity and at the same time provides diagnostic MSn data often critical for metabolite structural assignment. The linear ion trap was capable of dealing with the high chromatographic efficiencies and hence narrow peak widths associated with 1.7 microm particle-packed column separations. Polarity switching and data-dependent MSn data were generated with ease, and applied to the identification of metabolites found in human plasma.

Accurate identification and quantification of 11-nor-Δ9-tetrahydrocannabinol-9-carboxylic acid in urine drug testing: Evaluation of a direct high efficiency liquid chromatographic–mass spectrometric method

“…This technology offers highly efficient chromatography with significant advantages in resolution, speed and sensitivity for bioanalytical applications, particularly when coupled with high speed acquisition mass spectrometers [21]. This has already been used in pharmaceutical development [22,23], and for direct measurement of two serotonin metabolites in urine [24], and should also enable direct measurement of THCA in urine.…”

TopCount coupled to ultra-performance liquid chromatography for the profiling of radiolabeled drug metabolites in complex biological samples

“…Furthermore, the low system and gradient dwell volumes of UPLC instruments provide the opportunity to fully exploit the advantages of sub-2 m particle technology. Many reports have recently been published detailing the advantages of UPLC/MS for metabolite identification [12][13][14]. However, if UPLC is to truly compete with HPLC as a separation technique for the analysis of drug metabolites in complex matrices, then a suitable means of radiodetection is required, together with a full assessment of the robustness of UPLC to biological samples typically encountered in drug metabolism studies (e.g.…”

“…Although HPLC has become the linchpin of drug metabolite profiling and identification studies, the recent introduction of rugged small particle packed columns (<2 m), and the commercialisation of instrumentation which can withstand the ensuing high pressures, has lead to an increase in the application of socalled ultra-performance liquid chromatography (UPLC) in this field [12][13][14]. It is well documented that a decrease in particle size results in numerous benefits to the chromatographer, including a significant gain in efficiency, an increase in sensitivity and a concomitant increase in peak capacity per unit time in gradient separation [15].…”

Utilizing a −100°C microplate CCD Imager, yttrium silicate coated 384-microplates and ultra-performance liquid chromatography for improved profiling of radiolabeled drug metabolites in complex biological samples

“…Importantly, the use of radiolabel also facilitates the quantification of drug and metabolites in complex samples such as excreta and tissues, necessary to help define any species differences. Although high-performance liquid chromatography (HPLC) coupled to on-line radioactivity flow detectors (RFD) has been successfully used to separate and detect radiolabeled drug metabolites, the recent trend to smaller particle chromatography (<2 m) has lead to an increase in ultra-performance liquid chromatography (UPLC) methods in these applications [1][2][3]. It has been demonstrated in recent articles that UPLC methods, with inherent robustness, efficiency and resolution can be used for drug metabolite separation and identification, when coupled with fast scanning mass spectrometers [1][2][3].…”

“…Although high-performance liquid chromatography (HPLC) coupled to on-line radioactivity flow detectors (RFD) has been successfully used to separate and detect radiolabeled drug metabolites, the recent trend to smaller particle chromatography (<2 m) has lead to an increase in ultra-performance liquid chromatography (UPLC) methods in these applications [1][2][3]. It has been demonstrated in recent articles that UPLC methods, with inherent robustness, efficiency and resolution can be used for drug metabolite separation and identification, when coupled with fast scanning mass spectrometers [1][2][3]. The further coupling of UPLC to microplate scintillation counters (MSC), such as TopCount, has also been reported, thereby providing methods for both identification and quantification using off-line scintillation counting [4].…”