Interfacial Rheological and Emulsion Properties of Self-Assembled Cyclodextrin-Oil Inclusion Complexes.

To investigate the effect of the molecular size of alkanes and the cavity size of cyclodextrins (CDs) on the formation of interfacial host-guest inclusion complexes, the interfacial tension (IFT) of CD (α-CD, ß-CD, γ-CD) solutions against oils (hexadecane, dodecylbenzene) was determined by interfacial dilational rheology measurements. The results show that the "space compatibility" between CDs and oil molecules is crucial for the formation of interface host-guest inclusion complexes. Hexadecane with a smaller molecular size can form host-guest inclusion complexes with small cavities of α-CD and ß-CD, dodecylbenzene with a larger molecular size can form interfacial aggregates with the medium-sized cavity of ß-CD easily, and the polycyclic aromatic hydrocarbon molecules in kerosene can form inclusion complexes with the large cavity of γ-CD. The formation of interfacial inclusion complexes leads to lower IFT values, higher interfacial dilational modulus, nonlinear IFT responses to the interface area oscillating, and skin-like films at the oil-water interface. What's more, the phase behavior of Pickering emulsions formed by CDs with different oils is explored, and the phenomena in alkane-CD emulsions are in line with the results in dilatation rheology. The interfacial active host-guest structure in the kerosene-γ-CD system improves the stability of the Pickering emulsion, which results in smaller emulsion droplets. This unique space compatibility characteristic is of great significance for the application of CDs in selective host-guest recognition, sensors, enhanced oil recovery, food industries, and local drug delivery.

Systematic and comprehensive strategy for reducing matrix effects in LC/MS/MS analyses.

A systematic, comprehensive strategy that optimizes sample preparation and chromatography to minimize matrix effects in bioanalytical LC/MS/MS assays was developed. Comparisons were made among several sample preparation methods, including protein precipitation (PPT), liquid-liquid extraction (LLE), pure cation exchange solid-phase extraction (SPE), reversed-phase SPE and mixed-mode SPE. The influence of mobile phase pH and gradient duration on the selectivity and sensitivity for both matrix components and basic analytes was investigated. Matrix effects and overall sensitivity and resolution between UPLC technology and HPLC were compared. The amount of specific matrix components, or class of matrix components, was measured in the sample preparation extracts by LC/MS/MS with electrospray ionization (ESI) using both precursor ion scanning mode and multiple reaction monitoring (MRM). PPT is the least effective sample preparation technique, often resulting in significant matrix effects due to the presence of many residual matrix components. Reversed-phase and pure cation exchange SPE methods resulted in cleaner extracts and reduced matrix effects compared to PPT. The cleanest extracts, however, were produced with polymeric mixed-mode SPE (both reversed-phase and ion exchange retention mechanisms). These mixed-mode sorbents dramatically reduced the levels of residual matrix components from biological samples, leading to significant reduction in matrix effects. LLE also provided clean final extracts. However, analyte recovery, particularly for polar analytes, was very low. Mobile phase pH was manipulated to alter the retention of basic compounds relative to phospholipids, whose retention tends to be relatively independent of pH. In addition to the expected resolution, speed and sensitivity benefits of UPLC technology, a paired t-test demonstrated a statistically significant improvement with respect to matrix effects when this technology was chosen over traditional HPLC. The combination of polymeric mixed-mode SPE, the appropriate mobile phase pH and UPLC technology provides significant advantages for reducing matrix effects resulting from plasma matrix components and in improving the ruggedness and sensitivity of bioanalytical methods.

A study of ion suppression effects in electrospray ionization from mobile phase additives and solid-phase extracts.

Since the wide adoption of liquid chromatography/tandem mass spectrometry (LC/MS/MS), the ion suppression/enhancement phenomenon is the latest barrier to high-throughput analysis. This consequence of a nonoptimized analytical method can lead to adverse effects during quantitation (i.e. poor accuracy and precision). Previous papers have reported that ion suppression is a direct result of endogenous material present in biological samples. However, in the case of a solid-phase liquid chromatography/tandem mass spectrometry (SPE/LC/MS/MS) system, the measured result is the combination of several operating conditions and parameters. Little has been done to effectively monitor and/or choose optimized conditions for the complete sequence of extraction, clean up, separation and analysis. This paper describes a simple setup for quantification of ion suppression/enhancement. Several mobile phase additives, ion-pairing agents and SPE extracts were measured and compared against a standard reference. The results demonstrated that a clean up of plasma extracts based on ion exchange leads to minimal ion suppression/enhancement for the compounds that were investigated.

Analysis of a basic drug by on-line solid-phase extraction liquid chromatography/tandem mass spectrometry using a mixed mode sorbent.

Two on-line configurations using multiple 6- and 10-port valves were investigated for high-flow on-line extraction of a basic drug in rat plasma and human urine using a reversed-phase and a cation-exchange SPE sorbent. The first configuration studied was a single reversed-phase extraction cartridge (2.1 x 20 mm, 25 microm) using an optimized washing protocol. The results showed that up to 1.5 mL of sample (urine or plasma diluted 1:1) can be injected with limits of quantification (LOQs) as low as 100 pg/mL. The second configuration studied was the combination of a cation exchanger and a reversed-phase cartridge using at-column dilution. The results showed better LOQs than those obtained with the single cartridge at 10 pg/mL with the same injection volume. The mass spectrometer was operated in the multiple reaction monitoring (MRM) mode. All calibration curves gave an average of 5% relative standard deviation (RSD).

Performance of an ultra-low elution-volume 96-well plate: drug discovery and development applications.

Recently, sample preparation has been considered to be the major cause of bottlenecks during high-throughput analysis. With the assistance of robotic liquid handlers and the 96-well plate format, more samples can be prepared for subsequent liquid chromatography/tandem mass spectrometry (LC/MS/MS) analysis. Protein precipitation is still widely used despite potential loss of sensitivity or variable results due to ion suppression. The use of solid-phase extraction (SPE) clearly gives superior results but may not be as cost effective as protein precipitation due to the labor and material costs associated with the process. Here, a novel 96-well SPE plate is described that was designed to minimize the elution volume required for quantitative elution of analytes. The plate is packed with 2 mg of a high-capacity SPE sorbent that allows loading of up to 750 microL of plasma, while the novel design permits elution with as little as 25 microL. Therefore, the plate offers up to a 15-fold increase in sample concentration. The evaporation and reconstitution step that is typically required in SPE is avoided due to the concentrating ability of the plate. Examples of applications in drug discovery/development are shown and results are compared to protein precipitation. Excellent sensitivity and linearity are demonstrated.