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A lyophilization‒supercritical fluid extraction coupled with supercritical fluid chromatography‒quadrupole tandem mass spectrometry online method was developed for the determination of lipid mediators in breast cancer cells. Supercritical fluid extraction was applied to the cell samples for the first time due to the use of lyophilization. The conditions of supercritical fluid extraction and supercritical fluid chromatography‒quadrupole tandem mass spectrometry were investigated systematically. Under the optimized conditions, all the calibration curves for the lipid mediators showed good linearity (correlation coefficient > 0.99). The limits of detection and the limits of quantification were in the range of 0.190‒5.36 pg and 0.560‒16.2 pg, respectively. The recoveries were in the range of 70.3‒125%. The relative standard deviations of the precision ranged from 1.49‒18.7% and the accuracies were higher than 84%. Compared with liquid‒liquid extraction coupled with liquid chromatography and tandem mass spectrometry method, the present approach reduced the manual labor and obtained higher sensitivity as well as higher extraction recoveries for all 15 lipid mediators. Finally, the online method was applied to the quantification of lipid mediators in breast cancer cells and normal mammary epithelial cells. On the basis of the results, this lyophilization‒supercritical fluid extraction online coupled with supercritical fluid chromatography‒quadrupole tandem mass spectrometry method showed great promise in the analysis of lipid mediators in complex biological samples. [ABSTRACT FROM AUTHOR]

Background: Eicosanoids as inflammatory mediators take part in the regulation of disease progression. However, the application of serum eicosanoid in disease progression identification was still uncertain. Methods: Serum from 52 healthy volunteers, 34 enteritis patients and 55 colorectal cancer (CRC) patients were collected. Ultra-high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) was used to analyze the change of serum eicosanoids. Results: Of 158 eicosanoids, we found that lower levels of anti-inflammatory eicosanoids 13-HOTrE, 9-HOTrE, DHA, 11-HETE and 12-HHT were observed in enteritis and CRC group compared with healthy group, meanwhile the content of 5-iPF2α-VI as oxidative stress mediator in enteritis and CRC group was greater than that in healthy groups. Moreover, 9-HODE, 13-HODE, 12,13-diHOME, 8-HETE and 15-HETE were dramatically decrease in CRC group compared with non-CRC group. Additionally, the change of 5-, 12- and 15-HETE content in serum sample was associated with progression from healthy to enteritis, finally to CRC. No significant difference between serum eicosanoids and the expression of CerbB-2 and Ki67 were observed. Conclusion: Serum eicosanoids might be used as a possible biomarker for identifying among health, enteritis and CRC. [ABSTRACT FROM AUTHOR]

Sample preparation is very important in the chromatographic analysis due to its time consumed and deviations produced, therefore the problem of on-line sample preparation techniques is the front project in the field of analytical chemistry. In this paper, the progress of on-line sample preparation techniques in chromatographic analysis, including solid phase extraction, solid phase microextraction, liquid phase microextraction, membrane assisted extraction, field assisted extraction, gas phase extraction, thermal desorption and microfluidic chip is reviewed.