Sample preparation, electrospray ionization

Because plasma and urine are both aqueous matrixes, reverse-phase or polar organic mode enantiomeric separations are usually preferred as these approaches usually requires less elaborate sample preparation. Protein-, cyclodextrin-, and macrocyclic glycopeptide-based chiral stationary phases are the most commonly employed CSPs in the reverse phase mode. Also reverse phase and polar organic mode are more compatible mobile phases for mass spectrometers using electrospray ionization. Normal phase enantiomeric separations require more sample preparation (usually with at least one evaporation-to-dryness step). Therefore, normal phase CSPs are only used when a satisfactory enantiomeric separation cannot be obtained in reverse phase or polar organic mode. 

Desorption Electrospray Ionization (DESI). DESI is a novel gentle ionization method for surface analysis (Figure 2.6).[19,20] Like classical ESI, it operates at atmospheric pressure. No sample preparation is required. A solvent passes through the capillary of the electrospray source charged droplets are produced (primary ions) and they are directed to a solid sample. Their impact with the surface causes sample molecules to be ionized and... 

Desorption electrospray ionization DESI Spray Nonvolatile molecular ions Direct, preparation-free analysis of samples... 

J. Axelsson, A.-M. Hoberg, C. Waterson, P. Myatt, G. L. Shield, J. Varney, D. M. Haddleton, and P. J. Derrick. Improved Reproducibility and Increased Signal Intensity in Matrix-Assisted Laser Desorption/Ionization as a Result of Electrospray Sample Preparation. Rapid Commun. Mass Spectrom., 11(1997) 209-213. 

Bonfiglio, R., King, R. C., Olah, T. V, and Merkle, K. (1999). The effects of sample preparation methods on the variability of the electrospray ionization response for model drug compounds. Rapid Commun. Mass Spectrom. 13, 1175-1185. 

Fion, N., Gellon, J. O., Jensen, H., and Girault, H. H. (2003). On-chip protein sample desalting and preparation for direct coupling with electrospray ionization mass spectrometry. ]. Chromatogr. A 1003, 11-19. 

Direct analysis of solid samples or analytes present on solid surfaces without any sample preparation has always been a topic of interest. Desorption electrospray ionization (DESI) is an atmospheric pressure desorption ionization method introduced by Cooks et al., producing ions directly from the surface to be analyzed, which are then sampled with the mass spectrometer [22, 37]. DESI is based on charged liquid droplets that are directed by a high velocity gas jet (in the order of 300 m s ) to the surface to be analyzed. Analytes are desorbed from the surface and analyzed by mass spectrometer (Eig. 1.15). 

Mass spectrometry (MS) is widely used to ascertain the purity, total mass of the protein produced, and detect any covalent modifications (Cohen and Chait, 2001). Both electrospray ionization (ESI) and MALDI may be used although for intact proteins ESI has the advantage of being accurate to 1 Da. Using the simple protocol described in Protocol 2.11, the MS of whole protein samples can be readily automated without the need for sample preparation. This method has proved successful for the... 

Nanospray is a miniaturized version of electrospray. In the original setup of Wilm and Mann (8) it is utilized as an off-line technique using disposable, finely drawn (1 -gm tip), metallized glass capillaries to infuse samples at 10-30 nL/min flow rates. This allows more than 50 min analysis time with just a 1-pT sample. Due to the formation of much smaller droplets and the more effective ionization, there is often no need for LC separation, since the separation is accomplished in m/z or by MS/MS. However, limited reproducibility with respect to quantification and a more complex sample preparation can be seen as drawbacks. An on-line version for hyphenation with capillary and nano-LC as well as CE (slightly modified) is now commercially available. 

A newer approach for lipid analysis is electrospray ionization tandem mass spectrometry (ESI-MS/MS) (Welti et al., 2002). This method requires limited sample preparation and sample size to identify and quantify lipids. Fauconnier et al. (2003) used ESI-MS/MS to analyze phospholipid and galactolipid levels during aging of potato tubers. 

DESI Desorption electrospray ionization (DESI) is a recently developed technique that permits formation of gas-phase ions at atmospheric pressure without requiring prior sample extraction or preparation. A solvent is electrosprayed at the surface of a condensed-phase target substance. Volatilized ions containing the electrosprayed droplets and the surface composition of the target are formed from the surface and subjected to mass analysis (Takats et al., 2005 Wiseman et al., 2005 Kauppila et al., 2006). 

A sensitive and rapid chromatographic procedure using a selective analytical detection method (electrospray ionization-mass spectrometry in SIM mode) in combination with a simple and efficient sample preparation step was presented for the determination of zaleplon in human plasma. The separation of the analyte, IS, and possible endogenous compounds are accomplished on a Phenomenex Lima 5-/rm C8(2) column (250 mm x 4.6 mm i.d.) with methanol-water (75 25, v/v) as the mobile phase. To optimize the mass detection of zaleplon, several parameters such as ionization mode, fragmentor voltage, m/z ratios of ions monitored, type of organic modifier, and eluent additive in the mobile phase are discussed. Each analysis takes less than 6 min. The calibration curve of zaleplon in the range of 0.1-60.0 ng/ml in plasma is linear with a correlation coefficient of >0.9992, and the detection limit (S/N = 3) is 0.1 ng/ml. The within- and between-day variations (RSD) in the zaleplon plasma analysis are less than 2.4% (n = 15) and 4.7% (n = 15), respectively. The application of this method is demonstrated for the analysis of zeleplon plasma samples [14]. 

Miniaturized LC/MS formats based on micromachined chip-based electrospray emitters and ionization sources on silicon (Schultz et al., 2000 Licklider et al., 2000 Ramsey and Ramsey 1997 Xue et al., 1997) and plastic (Vrouwe et al., 2000 Yuan and Shiea, 2001, Tang et al., 2001) microchips is a proactive approach for scale-down platforms. Various micromachining processes are used to fabricate these devices. These microanalytical technologies would create integrated sample preparation and LC/MS applications. The potential benefits of such a system include reduced consumption of sample/reagents, low cost, and disposability. 

Because anurans, including L. splendida, often breed in water, the peptide components of the secretions of the paratoid and rostral glands were purified directly by HPLC. The active component was sequenced with the aid of electrospray ionization mass spectrometry (ESI-MS) and given the name splendipherin (17).76 The structure of this 25-amino acid peptide subsequently was confirmed by comparison with a synthetically prepared sample, which further established that all amino acids were of the L-configuration. 

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