Sulfoconjugates of sterols play important roles as neurosteroids, neurotransmitters, and ion channel ligands in health and disease. In most cases, sterol conjugate analysis is performed with liquid chromatography-mass spectrometry. This is a valuable tool for routine analytics with the advantage of direct sterol sulfates analysis without previous cleavage and/or derivatization. The complementary technique gas chromatography-mass spectrometry (GC-MS) is a preeminent discovery tool in the field of sterolomics, but the analysis of sterol sulfates is hampered by mandatory deconjugation and derivatization. Despite the difficulties in sample workup, GC-MS is an indispensable tool for untargeted analysis and steroid profiling. There are no general sample preparation protocols for sterol sulfate analysis using GC-MS. In this study we present a reinvestigation and evaluation of different deconjugation and derivatization procedures with a set of representative sterol sulfates. The advantages and disadvantages of trimethylsilyl (TMS), methyloxime-trimethylsilyl (MO-TMS), and trifluoroacetyl (TFA) derivatives were examined. Different published procedures of sterol sulfate deconjugation, including enzymatic and chemical cleavage, were reinvestigated and examined for diverse sterol sulfates. Finally, we present a new protocol for the chemical cleavage of sterol sulfates, allowing for simultaneous deconjugation and derivatization, simplifying GC-MS based sterol sulfate analysis.
A short isoprenylation protocol starting from non‐conjugated N‐Boc‐N‐(1,1‐dimethylallyl)hydrazones was developed utilising Thomson's traceless bond construction. This type of [3,3]‐sigmatropic rearrangement is catalysed by the Brønsted acid triflimide and liberates only gaseous by‐products. The required N‐Boc‐N‐allylhydrazine precursor is available in three steps starting from a known diazene using biocatalytic aldol addition and Tebbe olefination as key steps. Allylhydrazones are prepared via condensation with appropriate aldehydes. Scope and limitations of the [3,3]‐sigmatropic rearrangements are analysed.
This study evaluated the HiberGene Group B Streptococcus test, a CE-IVD-approved molecular assay for rapid detection of
Streptococcus agalactiae
[Group B Streptococcus (GBS)] in human clinical specimens. Performance of the assay in terms of specificity, sensitivity and genotype inclusivity was investigated using an extended specificity panel of 113 human and animal GBS isolates, and eight isolates from other streptococcal species, from the isolate collection of the German National Reference Center for Streptococci. Broth cultures were tested according to the manufacturer’s protocol, including lysis, heat denaturation and isothermal amplification. All 104/104 (100 %) human GBS isolates of nine serotypes (Ia, Ib, II, III, IV, V, VI, VII, VIII and non-typeable) were correctly identified by the assay as GBS. Additionally, 7/9 (78 %) GBS isolates from elephants were also correctly identified. Six isolates of other streptococcal species/subspecies (
Streptococcus anginosus
,
S. constellatus
,
S. castoreus
,
S. dysgalactiae
and
S. dysgalactiae equisimilis
) were correctly reported as negative. Two
S. pyogenes
(Group A Streptococcus) isolates gave invalid results. The HG Group B Streptococcus assay identified human GBS isolates in culture with 100 % sensitivity.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.