Cloud Point Extraction in the Determination of Drugs in Biological Matrices.

Cloud point extraction (CPE) is a simple, safe and environment-friendly technique used in the preparation of various samples. It was primarily developed for the assessment of environmental samples, especially analyzed for metals. Recently, this technique has been used in the extraction and determination of various chemical compounds (e.g., drugs, pesticides and vitamins), in various matrices (e.g., human plasma, human serum, milk and urine). In this review, we show that CPE is a reliable method of extraction and can be used in analytical laboratories in combination with other techniques that can be used in the determination of drugs and other chemicals in the human biological matrix. According to the literature, a combination of different methods provides good recovery and can be used in the simultaneous determination of many drugs in a single analysis. CPE can be optimized by changing its conditions (e.g., type of surfactant used, incubation temperature, pH and the addition of salts). In this review, we present the optimized CPE methods used in the determination of various pharmaceuticals and describe how the conditions affect the performance of extraction. This data might support future designing of the new CPE applications that are simple and more accurate. We compared CPE with other extraction methods and also showed the advantages and disadvantages of various extraction techniques along with a discussion on their environmental impact. According to the publications reviewed, it is obvious that CPE is an easy, safe, rapid and inexpensive method of extraction.

Matrix effect screening for cloud-point extraction combined with liquid chromatography coupled to mass spectrometry: Bioanalysis of pharmaceuticals.

Matrix effects are one of the most challenging issues in the analysis of complex samples using liquid chromatography coupled to mass spectrometry (LC-MS). Apart from the instrumental origin, these effects are also related to sample preparation. Cloud-point extraction (CPE) is rarely combined with LC-MS as it requires the use of surfactants which might interfere with droplet evaporation. Thus, they are suspected to cause a significant matrix effect (signal suppression). In this paper, 73 model drugs with different physicochemical properties were screened to analyse how susceptible LC-MS is to the absolute and relative matrix effect (ME) when coupled with CPE for measurements in human plasma. Three combinations of the surfactant Triton X-114 concentration (1.5% or 6%) and extraction temperature (40 or 55 °C) in six pH values gave over 1300 analyte-sample preparation condition pairs. A new term - surfactant effect (calculated for the standard solution) - allowed us to distinguish between the surfactant effect and that related to interferences from human plasma. The screening revealed that CPE combined with LC-MS is not related to a significant ME in the optimal pH of extraction. A significant absolute ME (<85% or>115%) was observed only for 25% of the analytes. Data processing (principal component analysis, classification trees, partial least squares-discriminant analysis) based on the extraction conditions and molecular descriptors helped to identify compounds prone to the matrix effect and speed up method development. A low surfactant concentration and low temperature decreased both the absolute and relative ME. pH of the extraction influenced only the relative ME. Low retention time reduced the risk of relative ME, whereas high polarity and the possibility of hydrogen bond formation minimized the occurrence of the surfactant effect and absolute ME. A significant relative ME (>15%) was observed only for 11% of the compounds, thus CPE merged with LC-MS allowed to measure drug concentrations in a reliable manner for majority of compounds. The presented approach may be further applied to other analytes and matrices.

Cloud-point extraction is compatible with liquid chromatography coupled to electrospray ionization mass spectrometry for the determination of antazoline in human plasma.

Cloud-point extraction (CPE) is attracting increasing interest in a number of analytical fields, including bioanalysis, as it provides a simple, safe and environmentally-friendly sample preparation technique. However, there are only few reports on the application of this extraction technique in liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis. In this study, CPE was used for the isolation of antazoline from human plasma. To date, only one method of antazoline isolation from plasma exists-liquid-liquid extraction (LLE). The aim of this study was to prove the compatibility of CPE and LC-ESI-MS/MS and the applicability of CPE to the determination of antazoline in spiked human plasma and clinical samples. Antazoline was isolated from human plasma using Triton X-114 as a surfactant. Xylometazoline was used as an internal standard. NaOH concentration, temperature and Triton X-114 concentration were optimized. The absolute matrix effect was carefully investigated. All validation experiments met international acceptance criteria and no significant relative matrix effect was observed. The compatibility of CPE and LC-ESI-MS/MS was confirmed using clinical plasma samples. The determination of antazoline concentration in human plasma in the range 10-2500ngmL(-1) by the CPE method led to results which are equivalent to those obtained by the widely used liquid-liquid extraction method.

Cloud-point extraction is compatible with liquid chromatography coupled to electrospray ionization mass spectrometry for the determination of bisoprolol in human plasma.

Cloud-point extraction (CPE) draws increasing interest in a number of analytical fields including bioanalysis, but combining CPE and LC-MS with electrospray ionization (ESI) in the determination of drugs in biological fluids such as plasma, serum or blood has not been reported so far. Bisoprolol was determined in human plasma by CPE using Trition X-114 as a surfactant and metoprolol as the internal standard. NaOH concentration, temperature and Trition X-114 concentration were optimized. All analyses were performed using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS). All validation experiments met international acceptance criteria and no significant matrix effect was observed. The compatibility of CPE and LC-ESI-MS/MS was confirmed using clinical plasma samples and appropriate statistical tests. The determination of bisoprolol concentration in human plasma in the range 1.0-70ngmL(-1) by the CPE method leads to the results which are equivalent to those obtained by the widely used liquid-liquid extraction method. The results revealed that a structural analogue may be an appropriate internal standard when CPE is used as the extraction technique. CPE offers significant practical advantages over the classical extraction methods, including a positive impact on the environment, therefore its wider application in future pharmacokinetic studies is justifiable.