Phase separation liquid-liquid extraction for the quantification of 8-iso-Prostaglandin F2 Alpha in human plasma by LC-MS/MS

Abstract

Summary Background. Reactive oxygen species (ROS) are produced in the body during normal metabolism by means of enzymes and non-enzymatic chemical reduction of molecular oxygen. In case of prevalence of ROS formation over their elimination, highly reactive free radicals can be accumulated and can cause multiple damages to the biomolecules and cells. Determination of isoprostanes in biological matrices is most often used to register free radical damage and requires selective, sensitive and specific techniques. Methods. This study presents the development and validation of an LC-MS/MS method for the determination of 8-iso-Prostaglandin F2α in human plasma utilising a modified liquid-liquid extraction procedure with phase separation. Results. Modified sample preparation procedure assured higher extraction yield, clear separation of organic layer from plasma water phase and protein precipitates, and better purified product for instrumental analysis. Linearity was validated in the range 0.1 – 5.0 µg/L with R2>0.996; normalised matrix varied between 86.0 and 108.3%, accuracy ranged from 90.4 % to 113.9% and precision both within-runs and between-runs was less than 7%. With a run time of 10 min, a throughput of over 50 samples per working day could be performed. Conclusion. The method meets all the current industrial validation criteria and allows the accurate and precise determination of 8-iso-PGF2α in human plasma at diagnostically significant concentration range.

Packed-Fiber Solid Phase-Extraction Coupled with HPLC-MS/MS for Rapid Determination of Lipid Oxidative Damage Biomarker 8-Iso-Prostaglandin F2α in Urine

“…The ME and normalized ME values were 95.4% and 104.2%, respectively. The data were in the 85–115% range, indicating that the influence of the urine matrix was well-controlled [ 25 ].…”

“…PS-electrospun nanofibers were originally introduced as the adsorbent material. Tomov et al proposed a liquid–liquid extraction (LLE) method to prepare plasma samples [ 25 ]; Im et al combined SPE, LLE, and derivatization for urine pretreatment [ 29 ], and Moral et al [ 30 ] proposed an SPE-after-derivatization method. But all these methods consumed a larger volume of organic solvent and required time-consuming nitrogen evaporation and redissolution steps to concentrate the sample.…”

“…However, GC/MS requires time-consuming and laborious prederivational treatment of the sample, which often produces artifacts and contamination [ 23 , 24 ]. Therefore, LC-MS has drawn more attention over the last two decades for its improved specificity and sensitivity, and much easier pretreatment [ 21 , 25 ].…”

Development and validation of an LC-MS/MS method for determination of 8-iso-prostaglandin f2 Alpha in human saliva

“…To achieve maximum sensitivity and specificity of the method, considering the low isoprostanes concentration in saliva (in ng/L range), we had to accomplish a very good chromatographic separation and optimal mass detector settings. Our previous experience with the core shell chromatographic column in determining isoprostanes in blood plasma (30) predetermined the use of the same column. Using a gradient elution, we achieved the cleanest and most symmetrical peaks, which was necessary for establishing a very good low limit of detection (LOD) and low limit of quantification (LLOQ) values, respectively 10 ng/L and 25 ng/L.…”

Oxidative Stress in the Oral Cavity before and After Prosthetic Treatment

“…The release of isoprostanes from membrane structures happens under the action of phospholipases and platelet-activating factor acetyl hydrolase (PAF-AH) [25]. The use of isoprostanes as a marker of oxidative stress level has two advantagestheir low chemical reactivity and presence in all biological fluids [26]. Furthermore, their local concentration may be used for assessment of the specific body system or area [27].…”