Automatic Supported Liquid Extraction (SLE) Coupled with HILIC-MS/MS: An Application to Method Development and Validation of Erlotinib in Human Plasma

Abstract

A novel bioanalytical method was developed and validated for the quantitative determination of erlotinib in human plasma by using the supported liquid extraction (SLE) sample cleanup coupled with hydrophilic interaction liquid chromatography and tandem mass spectrometric detection (HILIC-MS/MS). The SLE extract could be directly injected into the HILIC-MS/MS system for analysis without the solvent evaporation and reconstitution steps. Therefore, the method is simple and rapid. In the present method, erlotinib-d6 was used as the internal standard. The SLE extraction recovery was 101.3%. The validated linear curve range was 2 to 2,000 ng/mL based on a sample volume of 0.100-mL, with a linear correlation coefficient of > 0.999. The validation results demonstrated that the present method gave a satisfactory precision and accuracy: intra-day CV < 5.9% (<8.4% for the lower limit of quantitation, LLOQ) with n = 6 and the accuracy of 98.0–106.0%; inter-day CV < 3.2% (<1.5% for LLOQ) with n = 18 and the accuracy of 100.0–103.2%. A dilution factor of 10 with blank plasma was validated for partial volume analysis. The stability tests indicated that the erlotinib in human plasma is stable for three freeze-thaw cycles (100.0–104.5% of the nominal values), or 24-h ambient storage (100.0–104.8% of the nominal values), or 227-day frozen storage at both -20 °C (91.5–94.5% of the nominal values) and -70 °C (93.3–93.8% of the nominal values). The results also showed no significant matrix effect (<6.3%) even with direct injection of organic extract into the LC-MS/MS system. The validated method has been successfully applied to support a clinical study.

Recent developments in the chromatographic bioanalysis of approved kinase inhibitor drugs in oncology

“…The introduction of a 96-well format with a robotic liquid handling system for the analysis of erlotinib significantly improved the throughput of SLE. The methyl-tert butyl ether extract could be directly injected into the HILIC-MS/MS system for analysis without the solvent evaporation and reconstitution steps [91].…”

“…The most often used format in high throughput bioanalysis is the 96-well format microtiter plate. Most sample preparative methods can be employed on the 96-well format, and the aid of liquid handling robots makes processing the plates even faster, whilst requiring less human operation [91]. Of the reviewed TKI assays, some make use of the format for PP [31,72], SPE [21,92,94] and SLE [91].…”

“…Most sample preparative methods can be employed on the 96-well format, and the aid of liquid handling robots makes processing the plates even faster, whilst requiring less human operation [91]. Of the reviewed TKI assays, some make use of the format for PP [31,72], SPE [21,92,94] and SLE [91]. The 96-well format provides a universal format for analytical sample preparation, plates can be handled through devices like vacuum or pressurizing systems aiding in the flow through SPE or SLE columns or filters.…”

Separation and Determination of Process-Related Impurities of Erlotinib Using Reverse-Phase HPLC with a Photo-Diode Array Detector

“…A thorough literature survey has indicated several HPLC methods for the determination of erlotinib·HCl in pharmaceutical formulations as well as in biological fluids using ultra-violet and mass spectra detection. [3][4][5][6][7][8] The HPLC method was developed for the assay of erlotinib in the presence of degradation products. 9 The analysis of erlotinib and its metabolites is conducted in rat tissue sections by MALDI quadrupole time-of-flight mass.…”

A Rapid and Simple UHPLC-UV Method for Quantitative Determination of Erlotinib and Its Active Metabolite OSI-420 in Human Serum, and Its Application in a Non-Small Cell Lung Cancer Patient

“…Several analytical methods have been developed using liquid chromatography-tandem mass spectrometry (LC/MS/MS) [11][12][13][14][15][16][17] or high-performance liquid chromatography with ultraviolet detection (HPLC-UV) [3,18,19] to determine the concentrations of erlotinib and/or OSI-420 in human biological samples. LC/MS/MS methods are highly selective and sensitive, but due to their cost and complexity, they are not available in most hospital laboratories.…”