A new ionization method named surface-activated chemical ionization (SACI) has been realized. In this invention a commercially available atmospheric pressure chemical ionization (APCI) chamber, employed without any corona discharge (no-discharge APCI), has been modified with the insertion of a gold surface, leading to a significant improvement in the ionization efficiency. The ionization of the sample takes place by both gas-phase and surface-activated processes. This new ionization source is able to generate ions with high molecular mass and low charge states, leading to improved sensitivity and reduced noise. The new device has been tested in the analysis of some peptides. A comparison between the performance with and without the presence of the surface, and the optimization of the operating conditions (nebulizing gas flow, sample solution flow, pH of solution, and surface area), are reported and discussed.

In previous studies, the production of ions in an APCI source without any corona discharge was observed, and the intensity of the ion signals showed significant increases on placing a metallic surface at 45 degrees inside an orthogonal ion source. This method was named surface-activated chemical ionization (SACI). The present study was performed to investigate the mechanisms of ion production with or without the presence of the metallic surface, by varying instrumental parameters and the geometrical configuration. Approximate calculations show that, in the absence of corona discharge and of any additional surfaces, ions cannot be produced by collisional phenomena, because of their low kinetic energy, in the 10(-2) to 10(-3) eV range. Two alternative possibilities have been considered: the first takes into account that ions may originate by collision of neutral clusters of polar solvent molecules with the APCI source surfaces through clusterelectric effect. The second takes into account that the water dissociation constant k(w) is temperature dependent, passing from 10(-14.1669) at 20 degrees C to 10(-12.4318) at 90 degrees C. It means that the [H(+)] varies from 8.3 x 10(-8) to 6.1 x 10(-7) M going from 20 to 90 degrees C. Hence, at the high temperatures experimented in the APCI vaporizer, H(+) becomes available in solution in molar quantities analogous to those of analyte, and the protonation of the analyte itself can consequently occur. The activation of further ionization processes in the presence of the metallic surface can be reasonably attributed to interactions between gas-phase analyte molecules and solvent molecules adsorbed on the surface. Experiments performed with a thin layer of deuterated glycerol on the surface led to unequivocal results, i.e. the production of [M + D](+) ions of the analyte.

A novel approach, based on the use of atmospheric pressure chemical ionization ion trap mass spectrometry (APCI-ITMS) conditions, but without using corona discharge, was used to analyze peptides. The proposed method was applied to three standard peptides (bombesin, trityrosine and tyrosine-glycine-glycine) as well as peptides obtained through enzymatic digestion of two standard proteins (horse cytochrome c and horse myoglobin).

A new approach, based on the use of atmospheric pressure chemical ionization ion trap mass spectrometry (APCI-ITMS), but without a corona discharge, was investigated for application to creating and monitoring protein ions. It must be emphasized that APCI is not usually used in protein analysis. In order to verify the applicability of the proposed method to the analysis of proteins, two standard proteins (horse cytochrome c and horse myoglobin) were analyzed. A mixture of the two proteins was also analyzed showing that this novel approach, based on the use of APCI, can be used in the analysis of protein mixtures.

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The study of electron ionization (EI) induced decomposition patterns is usually undertaken by extensive metastable-ion investigations, mainly depending on spectra of the unimolecular decomposition of parent ions carried out in double-focusing instruments.By such an approach, in the last decades most of the definitive data on ion structures and on the mass spectrometric behaviour of many classes of compounds have been achieved. Unfortunately, single-quadrupole analysers do not give any definitive information on the unimolecular decomposition pathways of selected species; only by collision experiments in triple-stage quadrupolar systems can such data be obtained.','The ion-trap mass spectrometer3 (ITMS) offers a particularly interesting mass spectrometric approach due to its ability to store ions for further experiments, first of all being mass analysis. In particular, the use of a supplementary AC voltage (the so-called 'tickle' voltage) leads to effective collision-induced decompositions of preselected ionic species, even in cases where the energy deposition into the stored ionic species is lower than that usually observed in high-energy collision experiments.For these reasons ITMS seems to be highly promising for ion structure investigations as an alternative to the more complex (and more expensive) systems such as double-focusing sector instruments and triple-stage quadrupoles. In other words, what is usually obtained by the study of natural or collision-induced decomposition in multiple-stage analysers, is achieved in the ITMS by a simple, single-stage device by means of time separation of the different steps, namely electron ionization, ionic species isolation, collision of the preselected ions, mass analysis and detection of the collisioninduced decomposition products.Following our previous mass s ectrometric investigations of flavonoid cornpounds?'in this paper we will discuss the data obtained by ITMS on structure investigation of [M -HI+ ions of chalcone (2-propen-1-ona-1,3-diaphenyl) (l), whose EI mass spectrum has + Partially and preliminarily communicated at the European Meeting on Tandem Mass Spectrometry, Manchester UK, 9-11 July 1990.* Author to whom correspondence should be addressed.already been reported together with those of some of its derivatives.6-8 In such early papers the major fragmentation pathways and rearrangement processes were used to determine the type of chalcone and, in some cases, the position of substituents on the molecular skeleton. Some disagreement was expressed concerning the structure assignment of their [M -H]+ species.Ronayne and co-workers' postulated that the commonly observed intense [M -H]+ ion of chalcone was due to the loss of hydrogen from ring A in the resonance-stabilized flavylium ion structure a. The same mechanism was also proposed by Van

Through extensive studies of collisionally induced decompositions of different precursors by ion-trap mass spectrometry, the presence of a shadow region was demonstrated. If fragment ions fall into the stability diagram at a qz value of -0.78, their absolute intensities drop almost to zero. In order to test this phenomenon, different precursors and different collisionally produced fragment ions have been examined and in every case the fragment at qz= 0.78 disappeared, suggesting that such qr values can be considered as black holes in the microcosmos of the ion-trap mass spectrometer.Ion-trap mass spectrometry (ITMS) is, in our opinion, one of the most interesting mass spectrometric techniques to have been developed in recent years.' The potentialities of the instruments are currently under investigation: to date, their valid capabilities in mass analysis, ion isolation and multiple tandem mass spectrometry (MSIMS) experiments have been well p r~v e d .~.~ Mass range extension up to 25 000 Da, linked with collision yields very close to unity, make this instrument unique. We ourselves have tested the capabilities of ITMS in one of the most important Author to whom correspondence should be addressed. current mass spectrometric fields i.e., ion structure studies and isomer characterization based on collisional spectroscopy. In many cases exciting results were obtained.As has been described previously,' the equations of motion within the ion trap are described by second order differential equations (Mathieau equations), of the generalized form d2uwhere where U and V are the amplitudes of the DC and RF potentials applied at the ring electrode, ro the radius of the ring electrode, Q the angular frequency of the RF drive, m the mass of the ion. The a, and qz parameters are essential for describing the stability (trapping) or the instability (ejection) of an ion in the ion trap. Ions generated under EI conditions or from collisional experiments are usually detected at a,=O by massselective ejection of ions, occurring at q, = 0.908.'In MSIMS experiments if a third coordinate of the stability diagram is considered, so that the intensity of a daughter ion is plotted with respect to (az, qz) values, an unusual and unexpected behaviour has been observed by us: when the daughter ion falls in the range of ql values close to 0.78, its intensity drops dramatically, nearly to zero (Fig. 1). We will discuss here the results obtained by using different precursors and different daughter ions, and by variation of the main instrumental parameters, so as to obtain a phenomenological description of this strange behaviour.In Figure 2(a) the fragmentation pattern of chalcone, already studied by US,^ is reported together with its electron ionization mass spectrum (Fig. 2(b)). In Fig. 2(c) and (d) the daughter-ion spectrum of ionic species at rnlz 130 for tickle voltage of 220 mV and a tickle time of 15 ms is reported: this ion mainly decomposes by CO loss, giving rise to a daughter ion at mlz 102. Moving the parent ion position along the qz...

The new atmospheric pressure chemical ionization source, named surface-activated chemical ionization (SACI), has been used in conjunction with high-flow gradient chromatography to reduce the matrix effect. This high-flow gradient chromatography approach avoids the co-elution of analyte and biological matrix compounds that leads to a reduction in quantitation errors due to matrix effect. However, this approach cannot be employed with the classical electrospray ionization (ESI) source that usually works at low eluent flow (< 300 microL/min). SACI can work at high eluent flow (100-2000 microL/min) and can be employed in conjunction with high-flow gradient chromatography. The reduction in matrix effect in tacrolimus analysis in protein-precipitated blood samples, an important immunosuppressive agent for renal transplantation, is presented and discussed.