Sensitive mass spectrometric assay for determination of 15-deoxy-Δ12,14-prostaglandin J2 and its application in human plasma samples of patients with diabetes

Abstract

The determination of individual prostaglandins (PG) in humans is mainly performed in urine samples. The quantification of PGs in human plasma could improve the understanding of particular PG species under various physiological and pathological conditions. 15-Deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) is a dehydrated downstream product of PGD2 and is of high interest due to its recently discovered anti-inflammatory effects. Increasing availability of highly sensitive mass spectrometry allows the quantification of low abundant biomarkers like 15d-PGJ2 in human plasma samples. Herein, a sensitive LC-MS/MS method for the determination of 15d-PGJ2 was established. The method was validated according to the guidance of the American Food and Drug Administration and tested in plasma samples from patients with poorly controlled diabetes, considered to be a pro-inflammatory condition. Extraction of 15d-PGJ2 was achieved with an easy-to-use liquid-liquid extraction by ethyl acetate following a methanol precipitation. The lower limit of quantification was 2.5 pg mL−1 and linearity (R 2 = 0.998) was guaranteed between 2.5 and 500 pg mL−1 for 15d-PGJ2. Selectivity was assured by the use of two individual mass transitions (qualifier and quantifier). Precision and accuracy were validated in an inter- and intraday assay with a coefficient of variation below 11.8% (intraday) and 14.7% (interday). In diabetic patients with an HbA1C > 9%, increased plasma concentrations of 15d-PGJ2 compared to control plasma were measured. 15d-PGJ2 correlated negatively with the inflammation marker C-reactive protein. The developed LC-MS/MS method represents a new possibility to quantify 15d-PGJ2 with high specificity in human plasma samples. This may contribute to a better understanding of the potential anti-inflammatory effects of 15d-PGJ2 in severe long-term pro-inflammatory disorders like diabetes, cancer, or cardiovascular disease.Electronic supplementary materialThe online version of this article (10.1007/s00216-017-0748-1) contains supplementary material, which is available to authorized users.

Cannabidiol Regulates the Expression of Keratinocyte Proteins Involved in the Inflammation Process through Transcriptional Regulation

“…15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) was estimated using ultra-performing liquid chromatography tandem mass spectrometry (LCMS 8060, Shimadzu, Kyoto, Japan) [40]. PGF2α-d4 was used as an internal standard for quantification.…”

Acute fibrinolysis shutdown occurs early in septic shock and is associated with increased morbidity and mortality: results of an observational pilot study

“…Extraction of 6-keto-prostaglandin F1α (PG F1α) and 11-dehydro-thromboxane B 2 (11d-TX B 2 ) by liquid-liquid extraction was performed as described previously [10]. The quantification via LC-MS/MS was performed with minor modifications [10].…”

“…Extraction of 6-keto-prostaglandin F1α (PG F1α) and 11-dehydro-thromboxane B 2 (11d-TX B 2 ) by liquid-liquid extraction was performed as described previously [10]. The quantification via LC-MS/MS was performed with minor modifications [10]. Calibration range for each compound was as follows: PG F1α at 0.05, 0.1, 0.2, 0.4 and 0.8 pmol for each calibrator per injection; 11d-TX B 2 at 0.05, 0.1, 0.2, 0.4 and 0.8 pmol for each calibrator per injection.…”

A comparative study of small molecules targeting eIF4A

“…The large target spectrum of 15d-PGJ2 is borne out by a recent proteomic study of endothelial cells exposed to 5 µM 15d-PGJ2 for 4 h (far below the concentration at which effects on translation are reported), where 358 target proteins were identified, among which were many initiation (eIF4A1, eIF4G1, eIF4G3, eIF3B eIF3D, eIF3H, eIF3F) and elongation (EF1D, EF2, EF1G, mitochondrial EF-Tu) translation factors (Marcone and Fitzgerald 2013). The physiological concentrations of 15d-PGJ2 in body fluids has been reported to be in the nanomolar range-far lower than the concentrations required to inhibit translation in vitro (Morgenstern et al 2018).…”