RESUMEN
The bulbs of Fritillaria have been used for centuries as food and medicinal products in many Asian countries. Different Fritillaria species have distinct pharmacological effects despite of their similar appearances. Effective differentiation of Fritillaria species can avoid adulteration and is crucial to its clinical uses. In this paper, a hybrid method of matrix assisted laser desorption/ionization mass spectrometry and multivariate statistical analysis was developed for the rapid and reliable differentiation of Fritillaria species for the first time. Significantly different patterns for five Fritillaria species were obtained by MALDI-MS after instant sample extractions. Different groups of Fritillaria were confidently differentiated via an orthogonal partial least square model. In addition, a metabolomic taxonomy of five Fritillaria species was obtained based on MALDI-MS data.
Asunto(s)
Medicamentos Herbarios Chinos/análisis , Fritillaria/química , Metabolómica/métodos , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción/métodos , Contaminación de Medicamentos/prevención & control , Medicamentos Herbarios Chinos/química , Medicamentos Herbarios Chinos/normas , Fritillaria/clasificación , Fritillaria/metabolismo , Aprendizaje Automático , Análisis Multivariante , Raíces de Plantas/química , Raíces de Plantas/metabolismo , Análisis de Componente Principal , Reproducibilidad de los ResultadosRESUMEN
Peptides adducted with different divalent Group IIB metal ions (Zn(2+), Cd(2+), and Hg(2+)) were found to give very different ECD mass spectra. ECD of Zn(2+) adducted peptides gave series of c-/z-type fragment ions with and without metal ions. ECD of Cd(2+) and Hg(2+) adducted model peptides gave mostly a-type fragment ions with M(+â¢) and fragment ions corresponding to losses of neutral side chain from M(+â¢). No detectable a-ions could be observed in ECD spectra of Zn(2+) adducted peptides. We rationalized the present findings by invoking both proton-electron recombination and metal-ion reduction processes. As previously postulated, divalent metal-ions adducted peptides could adopt several forms, including (a) [M + Cat](2+), (b) [(M + Cat - H) + H](2+), and (c) [(M + Cat - 2H) + 2H](2+). The relative population of these precursor ions depends largely on the acidity of the metal-ion peptide complexes. Peptides adducted with divalent metal-ions of small ionic radii (i.e., Zn(2+)) would form predominantly species (b) and (c); whereas peptides adducted with metal ions of larger ionic radii (i.e., Hg(2+)) would adopt predominantly species (a). Species (b) and (c) are believed to be essential for proton-electron recombination process to give c-/z-type fragments via the labile ketylamino radical intermediates. Species (c) is particularly important for the formation of non-metalated c-/z-type fragments. Without any mobile protons, species (a) are believed to undergo metal ion reduction and subsequently induce spontaneous electron transfer from the peptide moiety to the charge-reduced metal ions. Depending on the exothermicity of the electron transfer reaction, the peptide radical cations might be formed with substantial internal energy and might undergo further dissociation to give structural related fragment ions.
Asunto(s)
Metales Pesados/química , Péptidos/química , Secuencia de Aminoácidos , Aminoácidos/química , Cationes/química , Radicales Libres/química , Espectrometría de MasasRESUMEN
Series of doubly and triply protonated diarginated peptide molecules with different number of glutamic acid (E) and asparagine (N) residues were analyzed under ECD conditions. ECD spectra of doubly-protonated peptides show a strong dependence on the number of E and N residues. Both the backbone cleavages and hydrogen radical (H*) loss from the charge-reduced precursor ions ([M+2H](+*)) were suppressed as the number of E and N residues increases. A strong inhibition of the backbone cleavages and H* loss from [M+2H](+*) was found for peptides with 6E residues (or 4E + 2N residues). The results obtained using these model peptides were re-confirmed by analyzing N-arginated Fibrinopeptide-B (i.e., REGVNDNEEGFFSAR). In contrast to the N-arginated peptide, ECD of the doubly-protonated Fibrinopeptide-B and its analogues show extensive backbone cleavages leading to series of c- and z-ions ( approximately 80% sequence coverage). Based on these results, it is believed that peptide ions with all surplus protons sequestered in arginine-residues would show enhanced stability under ECD conditions as the number of acid-residue increases. The suppression of backbone cleavages and H* loss from [M+2H](+*) are presumably attributed to the low reactivity of the charge-reduced precursor ions. One of the possible hypothesis is that diarginated E-rich peptides may contain hydrogen bonds between carbonyl oxygen of E side chains and backbone amide hydrogen. These hydrogen bonds would provide extra stabilization for [M+2H](+*). This is the first demonstration of natural structural motifs in peptides that would inhibit the backbone fragmentation of the charge-reduced peptide ions under ECD conditions.
Asunto(s)
Secuencias de Aminoácidos , Iones/química , Espectrometría de Masas/métodos , Péptidos/química , Secuencia de Aminoácidos , Fibrinopéptido B/química , Datos de Secuencia Molecular , ProtonesRESUMEN
Electron capture dissociation (ECD) of a series of custom-synthesized oligonucleotide pentamers was performed in a Fourier-transform mass spectrometer with a conventional filament-type electron gun. Dissociation of oligonucleotide ions by electron capture generates primarily w/d-type and z/a-type ions with and without the loss of a nucleobase fragment ions. Minor yields of radical [z/a + H]. fragment ions were also observed in many cases. It is interesting to note that some nucleoside-like fragment ions and protonated nucleobase ions (except thymine-related nucleobases and nucleoside-like fragments) were observed in most ECD spectra. The formation of these low-mass fragment ions was tentatively attributed to the secondary fragmentation of the radical [z + H]. fragment ions. From the ECD tandem mass spectra of a series of C/T based binary oligonucleotide ions, including d(CTCTC), d(CTTTC), d(TCCCT), d(CCCCT), and d(TCCCC), it was clearly demonstrated that the formation of many sequence ions was sensitive to the position of cytosine (or the position of charge carrier). The findings of this work support a notion that the ECD of protonated oligonucleotide molecules is charge-directed with the electron being captured by the protonated nucleobase.
Asunto(s)
Electroquímica/métodos , Espectrometría de Masas/métodos , Oligodesoxirribonucleótidos/química , Secuencia de BasesRESUMEN
Alkaloid profiles in Stemona tuberosa were found to be highly variable. Six Stemona alkaloids isolated from the plant were subjected to on-line high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) and tandem mass spectrometry (MS/MS) analyses. Their fragmentation patterns and products were useful for their characterization. The LC/MS fingerprints of these alkaloids, though variable among samples, could provide an overall characterization of the authenticity and quality of this species and help to differentiate it from S. japonica and S. sessilifolia, as all three species are recognized as genuine sources of the herb Radix Stemonae in the Pharmacopoeia of the People's Republic of China.