rFSH in medically assisted procreation: Evidence for ovarian follicular hyperplasia and interest of mass spectrometry to measure 17-hydroxyprogesterone and Δ4-androstenedione in serum.

Ovarian monitoring requires the determination of serum estradiol and progesterone levels. We investigated whole follicular steroidogenesis under rFSH in medically assisted procreation (MAP: 26 IVF, 24 ICSI) compared to 11 controls (IUI). Estrone, estradiol, Δ4-androstenedione, testosterone, progesterone and 17-hydroxyprogesterone were measured by immunoassay and mass spectrometry except for estrogens. At the start of a spontaneous or induced cycle, steroids levels fluctuated within normal ranges: estradiol (314-585 pmol/L), estrone (165-379 pmol/L) testosterone (1.3-1.6 nmol/L), Δ4-androstenedione (4.5-5.6 nmol/L), 17-hydroxyprogesterone (2.1-2.2 nmol/L) and progesterone (1.8-1.9 nmol/L). 17-hydroxyprogesterone, Δ 4-androstenedione and estradiol predominated. Then estradiol and oestrone levels rise, but less markedly for oestrone in IUI. In MAP, rFSH injections induce a sharp increase in estrogens associated with a rise in 17-hydroxyprogesterone and Δ4-androstenedione levels, disrupting oestrogen/androgen ratios. rFSH stimulation induces an ovarian hyperplasia and Δ4pathway which could become abnormal. Determining 17-hydroxyprogesterone and Δ4-androstenedione levels with LC-MS/MS may therefore be useful in managing recurrent MAP failures.

Ultra high performance liquid chromatography-quadrupole-time of flight analysis for the identification and the determination of resveratrol and its metabolites in mouse plasma.

Resveratrol is a polyphenol that has numerous interesting biological properties, but, per os, it is quickly metabolized. Some of its metabolites are more concentrated than resveratrol, may have greater biological activities, and may act as a kind of store for resveratrol. Thus, to understand the biological impact of resveratrol on a physiological system, it is crucial to simultaneously analyze resveratrol and its metabolites in plasma. This study presents an analytical method based on UHPLC-Q-TOF mass spectrometry for the quantification of resveratrol and of its most common hydrophilic metabolites. The use of (13)C- and D-labeled standards specific to each molecule led to a linear calibration curve on a larger concentration range than described previously. The use of high resolution mass spectrometry in the full scan mode enabled simultaneous identification and quantification of some hydrophilic metabolites not previously described in mice. In addition, UHPLC separation, allowing run times lower than 10 min, can be used in studies that requiring analysis of many samples.

Residual glutaraldehyde levels in fiberoptic endoscopes: measurement and implications for patient toxicity.

Most gastroenterology societies recommend glutaraldehyde for fiberoptic endoscope disinfection. However, glutaraldehyde toxicity has been suspected in patients examined with endoscopes disinfected with this compound. The aim of our study was to determine the residual levels of glutaraldehyde in fiberoptic endoscopes after either manual or automatic disinfection and to evaluate the extent of toxicity. Furthermore, the procedures for disinfection currently performed by the department were compared with the new French guidelines. We used both manual and automatic disinfection procedures and flushed sterile distilled water through the lumens of endoscopes before use. Residual glutaraldehyde levels were determined using liquid chromatography coupled to spectrophotometric detection. In a total of 92 measurements it was found that residual glutaraldehyde levels were higher and more variable after manual disinfection (< 0.2-159.5 mg/L) than after automatic disinfection (< 0.2-6.3 mg/L). We conclude that local procedures for disinfection need to be improved to conform to the new French guidelines. Since thresholds for the toxic dose of glutaraldehyde and international norms for levels of residual glutaraldehyde in equipment have not been defined, additional studies combining accurate measurements in fiberoptic endoscopes and clinical observations of endoscopy patients will be required to draw more definitive conclusions.

Fast specific separation and sensitive quantification of bactericidal and sporicidal aldehydes by high-performance liquid chromatography: example of glutaraldehyde determination.

This article describes the design and the validation of the HPLC determination of glutaraldehyde at g/l and mg/l concentrations, after derivatization by 2,4-dinitrophenylhydrazine and using the external standard method. At low concentrations, the reaction mixture needs to be heated and a weight ratio of 500 for the 2,4-dinitrophenylhydrazine reagent and the glutaraldehyde ensures a linear calibration curve. In contrast, high concentrations do not require heating of the reaction mixture and a weight ratio of 32 proved to be sufficient. The optimized HPLC method has been validated for both ranges of concentrations. Between 1.25 and 10 mg/l, the content can be determined by the external standard method, with a repeatability of 0.5%. The detection limit is 0.2 mg/l. Between 0.31 and 2.5 g/l, the content can also be determined by the external standard method, with a repeatability of 0.4%. Finally, statistical analysis has demonstrated that aqueous solutions of glutaraldehyde are stable for at least three days at 4 degrees C within the mg to g range.