Distinction of chiral penicillamine using metal-ion coupled cyclodextrin complex as chiral selector by trapped ion mobility-mass spectrometry and a structure investigation of the complexes.

Penicillamine (Pen) is a common chiral drug that is obtained from penicillin. Between the two enantiomers of Pen, only D-Pen can be used to treat cystinuria and rheumatoid arthritis while L-Pen is toxic. Therefore, it requires great efforts for the research of the rigorous analysis and distinction of the two enantiomers. The non-covalent combination of chiral molecules and chiral selectors (CSs) has been proved as a unique strategy for chiral distinction by ion mobility spectrometry in coupling with -mss spectrometry (IM-MS). Here, we developed a simple method to distinguish D, L-Pen by using special CSs for IM-MS separation. The CSs utilized here include cyclodextrins (CD) and linear chain oligosaccharides plus metal ions. We found that non-covalent complexes [Pen+ß-CD + Li]+ could be easily formed by electrospray ionization of the mixture of the solution, and the chirality of Pen could be effectively recognized by measuring their mobilities due to the different collision cross collision sections of [D-Pen+ß-CD + Li]+ and [L-Pen+ß-CD + Li]+. A detailed analysis of [Pen+ß-CD + Li]+ was then conducted by the optical rotation measurements and NMR experiments to reveal their structural differences. Furthermore, DFT calculation showed the differences of molecular conformation between the complexes. The results provide a new powerful method for fast analysis and recognition of chirality of Pen compounds by IM-MS.

Direct distinction of ibuprofen and flurbiprofen enantiomers by ion mobility mass spectrometry of their ternary complexes with metal cations and cyclodextrins in the gas phase.

Enantiomeric drugs are widely used and play important roles in pharmaceuticals. Ion mobility spectrometry coupled with mass spectrometry technology provides a unique method for distinguishing the enantiomeric drugs, enantiomeric identification, and quantitation in the gas phase. In this study, enantiomeric molecules of ibuprofen and flurbiprofen were clearly recognized by forming host-guest complex ions using trapped ion mobility time-of-flight mass spectrometry. Ternary complex ions can be produced easily by electrospray ionization of the mixed solutions of ibuprofen, cyclodextrins, and CaCl2 , LiCl, or NaCl, as well as flurbiprofen, cyclodextrins, and CaCl2 . The relative contents of different chiral ibuprofens in a mixed solution were also quantitatively measured. This new method is a simple, effective, and a convenient enantioselective analysis method.

Discrimination of Aminobiphenyl Isomers in the Gas Phase and Investigation of Their Complex Conformations.

The analysis of positional isomers is of great significance because their different chemical properties but similar structures make separation difficult. In this work, a simple method for simultaneously discriminating three positional isomers of 2-aminobiphenyl (2-ABP), 3-ABP, and 4-ABP was studied by ion mobility spectrometry (IMS) and quantum mechanical calculations at the molecular level. In the experiments, three ABP isomers were mixed with α-, ß-, and γ-cyclodextrins (CD), and the IMS results show that the three ABP isomers were clearly recognized by the formed complex of [α-CD + ABP + H]+ via measuring their IMS, in which the different ion mobilities of 1.515, 1.544, 1.585 V·s·com-2 with the collision cross sections (CCS) of 307.3, 312.5, 320.8 Å2 were obtained for [α-CD + 2-ABP + H]+, [α-CD + 3-ABP + H]+, and [α-CD + 4-ABP + H]+, respectively. Collision induced dissociation analysis of the three [α-CD + ABP + H]+ isomer complexes were further studied, indicating that the same fragmentation process required different collisional energies, and the greater the CCS for the [α-CD + ABP + H]+ with looser structure and the smaller energy required. Besides, the favorable conformation and the CCS value of the different [CD + ABP + H]+ isomer complexes were measured via quantum mechanical calculations to detail their intermolecular interactions. It revealed that the intermolecular binding between 2-ABP and α-CD is different from that of 3- and 4-ABP, resulting in different molecular conformations and CCS, and the interaction modes of ABP with ß-CD are similar to that with γ-CD, which are very consistent with the experimental observations. Finally, relative quantification of the method was performed, and satisfactory linearity with correlation coefficients (R2) greater than 0.99 was obtained. This method for isomer discrimination and conformation analysis possesses the advantages of simplicity, sensitivity, cost-effectiveness, and as such it may be widely applied in chemistry and pharmaceutical sciences.

Mass filter with phase modulation of radio frequency voltage.

In this work, the parametric quadrupole resonance caused by the phase modulation of waveform potential is studied. Based on analytical and numerical description of the ion motion in the quadrupole mass filter with the phase modulation, a stability island is found with good ion optical properties such as high ion transmission efficiency (16%), high mass resolution (peak width measured at 10% of peak height, R0.1 = 6000), the required separation time (100-150 radio frequency [RF] cycles), and good peak shape of trapezoid form. Furthermore, the analysis of the frequency spectrum of the applied potential and the quadrupole mass filter (QMF) acceptance are also presented; a resonance frequency is found from this spectrum. Finally, a suitable stability X-islands with relative modulation frequency, ν = 2 ± ß , ß = 1 P , 10 ≤ P ≤ 40 , is established and studied in detail. Here, ß is the imaginary part of the characteristic exponent of stable solutions to the Mathieu equation.

Investigation of noncovalent interactions between peptides with potential intrinsic sequence patterns by mass spectrometry.

RATIONALE: The conformation of a protein largely depends on the interactions between peptides. Specific and intrinsic sequence peptide patterns, such as DNA double helix backbones, may be present in proteins. A computational statistical deep learning method has supported this assumption, but it has not been experimentally proven. Mass spectrometry, as a fast and accurate experimental method, could be used to evaluate the interaction of biomolecules. The results would be of great value for further study of the mechanism of protein folding. METHODS: Several potential intrinsic peptides were chosen by the deep learning method, including seven groups of pentapeptides and five groups of nonapeptides. The noncovalent interactions between mixed polypeptides were investigated by electrospray ionization mass spectrometry (ESI-MS) in full-scan and collision-induced dissociation (CID) modes. Molecular dynamics and molecular mechanics Poisson-Boltzmann surface area (MD-MM/PBSA) analyses were also performed to support the results. RESULTS: The ESI-MS spectra showed that 11 of the 12 groups of mixed polypeptides formed binary and ternary complexes with relatively high stability. The binding between nonapeptide groups was stronger than that between pentapeptide groups according to the relative intensity. The binding energies calculated by the MM/PBSA binding energy tool also provided strong evidence for the combination of the complexes. Electrostatic interactions, hydrophobic interactions, and van der Waals forces were thought to stabilize the complexes according to the binding models. CONCLUSIONS: The results implied the formation of stable complexes between polypeptides and identified their noncovalent interactions, proving that specific sequences and combinations with relatively strong binding ability exist in potential intrinsic sequences of peptides in protein structures.

Simultaneous enrichment and analysis of tobacco alkaloids by microextraction coupled with mass spectrometry using a poly (N-isopropyl-acrylamide-co-divinyl-benzene-co-N, N'-methylene diacrylamide) monolithic column.

Herein, we realized the simultaneous online detection of six tobacco alkaloids (TAs) by in-tube solid-phase microextraction (In-tube SPME) coupled with mass spectrometry by a rapid, sensitive, and matrix effect-free method requiring no chromatographic separation and only minimal sample pre-treatment. A poly (N-isopropylacrylamide-co-divinylbenzene-co-N, N'-methylenediacrylamide) [Poly (NIPAAm-co-DVB-co-MBAA)] monolithic column was designed according to the chemical structures of selected TAs and used as an extraction medium engaging in hydrophobic, π-π, and hydrogen bonding interactions with analytes, allowing them to be effectively extracted. A number of important parameters were systematically optimized to achieve maximal extraction efficiency. The ion intensity of the TAs signals obtained by in-tube SPME-MS were higher than the direct MS mode by about 400 folds with the signal-to-noise ratio improved by 2-7 folds. The detection limits of the six TAs were determined as 1.99-4.06 ng g-1, with good linearity with correlation coefficients exceeding 0.99 obtained under optimal extraction conditions. Besides, TA recoveries in cigarette tobacco spiked at three concentration levels were in the range of 76.4-100.2%, and the corresponding RSDs (n = 5) were obtained as 4.32-7.16%. The extraction performance of the poly (NIPAAm-co-DVB-co-MBAA) monolithic column was well reproducible, with intra- or inter-day precision RSDs determined not to exceed 7.38%. Finally, no marked matrix effects were observed when the developed method was applied to the analysis of both high-abundance and trace-level TAs in practical samples, and the above technique was therefore concluded to be well suited for the detection of TAs in cigarette tobacco or other products.

Poly (methacrylic acid-co-diethenyl-benzene) monolithic microextraction column and its application to simultaneous enrichment and analysis of mycotoxins.

A poly (methacrylic acid-co-divinyl-benzene) [poly (MAA-co-DVB)] monolithic column was specially prepared according to the chemical structures of the three mycotoxins of aflatoxin B1, zearalenone and sterigmatocystin, and used for in-tube solid-phase microextraction (in-tube SPME) of the selected mycotoxins. The poly (MAA-co-DVB) monolithic column was characterized in detail, and exhibited effective hydrophobic, π-π, and hydrogen bonding interactions towards the target analytes. By coupling the poly (MAA-co-DVB) monolithic column-based in-tube SPME with high-performance liquid chromatography, a simple, sensitive and matrix effect-free method for determination of mycotoxins was developed. Some important parameters related to extraction efficiency were optimized systematically. Under the optimal conditions, the enrichment factors for the three mycotoxins ranged from 71.9 to 98.7. Recoveries of mycotoxins in rice grains with three spiked concentration levels were in the range of 78.0-102.8%, with satisfactory relative standard deviations (n = 4) of 2.96-4.74%. Good linearity was obtained with correlation coefficients larger than 0.99, and the detection limits (S/N = 3) of the three mycotoxins were between 0.69 and 2.03µgkg-1. The developed poly (MAA-co-DVB) monolithic column based in-tube SPME-HPLC method was successfully used to sensitively determine trace mycotoxins in practical samples of rice grain without interference peaks, indicating that it is promising for the detection of trace mycotoxins in complex grain samples.

Rapid fabrication of ionic liquid-functionalized monolithic column via in-situ urea-formaldehyde polycondensation for pressurized capillary electrochromatography.

A novel strategy for rapidly fabricating ionic liquid (IL)-bonded multifunctional monolithic stationary phase has been developed by an in-situ polycondensation of urea-formaldehyde (UF) and a lab-made acylamino-functionalized IL (1-acetylamino-propyl-3-methylimidazolium bromide, [AAPMIm]Br). Two polycondensation processes of UF with 1-amino-propyl-3-methylimidazolium bromide or [AAPMIm]Br were evaluated. Several parameters including mass ratio of urea-formaldehyde, amount of [AAPMIm]Br, polycondensation time and reaction temperature were optimized, and the [AAPMIm]Br-bonded monolithic stationary phase could be rapidly synthesized in 10min with a satisfactory permeability and mechanical stability. Used for pressurized capillary electrochromatography (pCEC), a typical hydrophilic interaction (HI) retention could be obtained in the resultant [AAPMIm]Br-bonded monolith when the content of acetonitrile (ACN) in mobile phase exceeded 20%. Multiple retention mechanisms such as hydrophilic interaction (HI), hydrogen bond (HH), anion-exchange and cation-exclude interactions, were acheived in the [AAPMIm]Br-bonded monolith. Various polar compounds including phenols, benzoic acid and its homologues, and enkephalins have been well separated and thus demonstrated a satisfactory separation performance of the obtained monolith. A facile access is lighted for rapid preparation of ionic liquid-bonded monoliths with multiple retention mechanisms for pCEC.

ETME, a novel ß-elemene derivative, synergizes with arsenic trioxide in inducing apoptosis and cell cycle arrest in hepatocarcinoma cells via a p53-dependent pathway.

Arsenic trioxide (ATO) has been identified as an effective treatment for acute promyelocytic leukemia (APL) but is much less effective against solid tumors such as hepatocellular carcinoma (HCC). In the search for ways to enhance its therapeutic efficacy against solid tumors, we have examined its use in combination with a novel derivative of ß-elemene, N-(ß-elemene-13-yl)tryptophan methyl ester (ETME). Here we report the effects of the combination on cell viability, apoptosis, the cell cycle and mitochondria membrane potential (MMP) in HCC SMMC-7721 cells. We found that the two compounds acted synergistically to enhance antiproliferative activity and apoptosis. The combination also decreased the MMP, down-regulated Bcl-2 and pro-proteins of the caspase family, and up-regulated Bax and BID, all of which were reversed by the p53 inhibitor, pifithrin-α. In addition, the combination induced cell cycle arrest at the G2/M phase and reduced tumor volume and weight in an xenograft model of nude mice. Overall, the results suggest that ETME in combination with ATO may be useful in the treatment of HCC patients particularly those unresponsive to ATO alone.