MEPS as a Rapid Sample Preparation Method to Handle Unstable Compounds in a Complex Matrix: Determination of AZD3409 in Plasma Samples Utilizing MEPS-LC-MS-MS

Abstract

The aim of the present investigation is to develop a simple, fast, and sensitive method for the determination of a new candidate drug, AZD3409, in rat, dog, and human plasma samples. AZD3409 is stable in aqueous solutions at low pH (< 4) but not in whole blood or in plasma. In rat plasma at 25 degrees C, more than 90% of the compound is degraded within 40 min. When 20 mg of NaF and 50 microL of protease inhibitor cocktail are added to 1.0 mL of rat blood, AZD3409 is stable for up to about 90 min. Due to the instability of AZD3409, microextraction in packed syringe (MEPS) is used as an online and fast sample-preparation method, followed by liquid chromatography-tandem mass spectrometry (LC-MS-MS) for the quantitation of this compound in plasma samples. In MEPS, the sampling sorbent is 1 mg of polystyrene polymer packed in a 250-microL syringe. When the plasma sample (50-250 microL) is withdrawn through the syringe by an autosampler, the analyte is adsorbed to the solid phase. The analyte is then eluted with an organic solvent such as methanol or the LC mobile phase (20-50 microL) directly into the instrument's injector. MEPS is rapid and easy to use. The lower limit of quantitation for AZD3409 is established to be 0.024 microM. The accuracy of the quality-control samples ranged from 89% to 102%, and the precision (C.V.%) had a value of 11-16% for the plasma samples. The calibration curve in plasma is obtained in the concentration range 0.022-9.0 microM. The coefficients of determination (R2) for plasma samples were> or = 0.998 for all runs. The present method is used for the analysis of rat and dog plasma samples.

A review of current trends and advances in modern bio-analytical methods: Chromatography and sample preparation

“…Typical examples include silica-based (C2, C8, C18), RAM or MIPs sorbents [195]. MEPS can be connected on-line to an LC [196,[198][199][200][201][202][203][204] or a GC [195] without any modification of the instrument. Connection to a robot makes the method fully automated.…”

“…Thus, several drugs such as local anaesthetics and their metabolites [195,196]; the anticancer drugs roscovitine [198], olomoucine [199], cyclophosphamide [200] and busulfan [201]; the ␤-blocker drugs acebutolol and metoprolol [202]; the antidepressant drugs dopamine and serotonine [203] as well as anti-addictive methadone [204] have been extracted successfully from biological samples such as plasma, urine or blood.…”

Effectiveness of high-throughput miniaturized sorbent- and solid phase microextraction techniques combined with gas chromatography–mass spectrometry analysis for a rapid screening of volatile and semi-volatile composition of wines—A comparative study

“…The MEPS technique has been used to extract various analytes from biological samples. Several drugs such as local anaesthetics and their metabolites [21]; the anticancer drugs roscovitine [22], olomoucine [23], and busulfan [24]; the ␤-blocker drugs acebutolol and metoprolol [25]; the anti-depressant drugs dopamine and serotonine [26] as well as anti-addictive methadone [27] have been successfully extracted by MEPS from biological samples such as plasma, urine or blood.…”

The emerging technique, microextraction in packed syringes in separation science – Can it eliminate matrix effects and fit with LC/MS/MS strategy?

“…Microextraction in packed syringes (or sorbents), popularly known as MEPS, is being used in a number of recent research work [1][2][3][4][5][6][7][8][9][10]. Recently, a review appeared on the use of MEPS especially as it relates to the versatility of the new tool to provide an avenue for improved sample preparation to aid the speed, sensitivity and selectivity options provided by LC-MS/MS [11].…”

“…There is a strong notion that MEPS in particular provides differentiation from the regular SPE and therefore could replace the need of the analytical column in LC-MS/MS (i.e., directly used online with the MS detector). For instance, in the assay of a novel drug candidate, AZ3409, due to rapid degradation of the compound in spite of adding a stabilizer such as sodium fluoride, MEPS technique came in handy because it served the purpose of rapid online extraction and coupled directly with the injector interface of LC-MS/MS [1]. There has been a suggestion that MEPS incorporation into mainstream LC-MS/MS strategy may not only reduce sample handing time but also contribute for an improvement in detectable limits as in the case of methadone [3].…”