ABSTRACT
Urinary tetranor-PGDM is a useful diagnostic biomarker for food allergy which often affects infants. We attempted to extract and measure urinary tetranor-PGDM absorbed in polymer of diapers. We applied CaCl2 to the collected polymer, determined the adequate time length of shaking the polymer to release urine, and measured tetranor-PGDM in the extracted urine. This procedure provided high linearity and recovery rate in tetranor-PGDM measurement. We also found that urinary tetranor-PGDM was stable for 24 h at 4°C in diapers. This method can be useful to monitor the food allergic condition of non-toilet trained children.
Subject(s)
Diapers, Infant , Food Hypersensitivity/diagnosis , Liquid-Liquid Extraction/methods , Prostaglandin D2/analogs & derivatives , Biomarkers/urine , Calcium Chloride , Child, Preschool , Humans , Infant , Polymers , Prostaglandin D2/isolation & purification , Prostaglandin D2/urine , Temperature , Time FactorsABSTRACT
Prostaglandins (PG) and isoprostanes (iso-PG) may be derived through cyclooxygenase or free radical pathways and are important signaling molecules that are also robust biomarkers of oxidative stress. Their quantification is important for understanding many biological processes where PG, iso-PG, or oxidative stress are involved. One of the common methods for PG and iso-PG quantifications is LC-MS/MS that allows a highly selective, sensitive, simultaneous analysis for prostanoids without derivatization. However, the currently used LC-MS/MS methods require a multi-step extraction and a long (within an hour) LC separation to achieve simultaneous separation and analysis of the major iso-PG. The developed and validated for brain tissue analysis one-step extraction protocol and UPLC-MS/MS method significantly increases the recovery of the PG extraction up to 95 %, and allows for a much faster (within 4 min) major iso-PGE2 and -PGD2 separation with 5 times narrower chromatographic peaks as compared to previously used methods. In addition, it decreases the time and cost of analysis due to the one-step extraction approach performed in disposable centrifuge tubes. All together, this significantly increases the sensitivity, and the time and cost efficiency of the PG and iso-PG analysis.
Subject(s)
Brain Chemistry , Chromatography, High Pressure Liquid/methods , Isoprostanes/isolation & purification , Prostaglandins/isolation & purification , Tandem Mass Spectrometry/methods , Animals , Chromatography, High Pressure Liquid/economics , Dinoprostone/analysis , Dinoprostone/isolation & purification , Isoprostanes/analysis , Limit of Detection , Male , Mice , Prostaglandin D2/analysis , Prostaglandin D2/isolation & purification , Prostaglandins/analysis , Tandem Mass Spectrometry/economics , Time FactorsABSTRACT
Kinetic parameters of enzymatic and non-enzymatic transformations of [3H]prostaglandin H2 (PGH2) were determined; the maximum yield of [3H]PGD2 being obtained at the keobs/koobs ratio equal to 10. The two-stage enzymatic synthesis of [3H]PGD2 with high molar radioactivity (3.15 TBq/mmol) from [3H]arachidonic acid carried out. Its identity in properties to the natural PGD2 was shown in experiments on the inhibition of ADP-induced aggregation of thrombocytes and on enzymatic oxidation with 15-hydroxyprostaglandin dehydrogenase.
