This study reports the development and application of a liquid chromatography method coupled to electrospray tandem mass spectrometry (LC-MS/MS) for the identification and quantification of the five most common juvenile hormone (JH) homologs and methyl farnesoate (MF). The protocol allows the simultaneous analysis in a single LC run of JH I, JH II, JH III, JH III bisepoxide (JHB3) and JH III skipped bisepoxide (JHSB3). The identification of JHs is based on multiple reaction monitoring (MRM), using two of the most abundant fragmentation transitions for each hormone. Addition of deuterated JH III as an internal standard permits the absolute quantification of the different JHs. The JH homologs common structural features led to similar chromatographic behavior, as well as related fragmentation patterns, which facilitated the simultaneous detection of all the homologs in a single LC-MS/MS run. The protocol detects JHs in the low femtomole range, allowing often the analysis of JH in individual insects. Fragmentation of each of the JH homologs generates unique diagnostic ions that permitted the identification and quantification of JHs from samples of different species of Diptera, Lepidoptera, Heteroptera and Hymenoptera. Having a simple protocol, which can undisputedly determine the identity of the homologs present in a particular species, provides us with the opportunity to identify and quantify JHs existing in insects that are pests, vector of diseases or important research models.
- MeSH
- chromatografie kapalinová * MeSH
- Diptera chemie MeSH
- Heteroptera chemie MeSH
- Hymenoptera chemie MeSH
- juvenilní hormony analýza chemie MeSH
- Lepidoptera chemie MeSH
- tandemová hmotnostní spektrometrie * MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, N.I.H., Extramural MeSH
Antimicrobial peptides (AMP) which kill microbes by a fundamentally different mechanism of action than do traditional antibiotics are a new category of compounds capable of fighting resistant pathogens. The cationic AMP isolated from insects comprise a significant group among those 2000 peptides already registered in The Antimicrobial Peptide Database. The novel AMP include lucifensins – the insect defensins isolated from medicinal larvae of flies which are routinely used in maggot therapy and AMP ranking among -helical amphipathic peptides isolated from the venom of stinging hymenopterans. The AMP isolated from the venom show strong antimicrobial activities and low or moderate toxicity to eukaryotic cells. Systematic structure modification of these naturally occurring AMP resulted in their analogs with enhanced antimicrobial activities.
- Klíčová slova
- lucifensin,
- MeSH
- antibakteriální látky * izolace a purifikace MeSH
- antiinfekční látky izolace a purifikace MeSH
- defensiny * farmakokinetika farmakologie chemie izolace a purifikace klasifikace terapeutické užití MeSH
- farmaceutické databáze MeSH
- histatiny aplikace a dávkování farmakokinetika terapeutické užití MeSH
- Hymenoptera * chemie imunologie MeSH
- kationické antimikrobiální peptidy * farmakokinetika farmakologie chemie izolace a purifikace terapeutické užití MeSH
- lidé MeSH
- magaininy aplikace a dávkování farmakokinetika terapeutické užití MeSH
- mikrobiální testy citlivosti MeSH
- molekulární struktura MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH