Electrospray interface operation

The pump must provide stable flow rates from between 10 ttlmin and 2 mlmin with the LC-MS requirement dependent upon the interface being used and the diameter of the HPLC column. For example, the electrospray interface, when used with a microbore HPLC column, operates at the bottom end of this range, while with a conventional 4.6 mm column such an interface usually operates towards the top end of the range, as does the atmospheric-pressure chemical ionization (APCI) interface. The flow rate requirements of the different interfaces are discussed in the appropriate section of Chapter 4. 

The MS/MS response for each analyte must first be optimized on the specific instrument to be used. This is usually done by infusing a solution of the analyte into the HPLC mobile phase without a column present. The composition of the mobile phase should match that expected at the time of analyte elution within 25%. The instrument is first operated in the LC/MS mode, and the settings for the electrospray interface are... 

Owing to the anionic character of LAS, an electrospray ionisation (ESI) interface operated in negative ion mode is particularly attractive for the mass spectrometric detection of this surfactant type. Consequently, a great part of the atmospheric pressure ionisation-mass spectrometry (API-MS) work on LAS is devoted to the application of (— )-ESI-MS. 

For capillary zone electrophoresis (CZE) mass spectrometry coupling, another modification of an ESI interface has been developed. This interface uses a sheath flow of liquid to make the electrical contact at the CZE terminus, thus defining both the CZE and electrospray field gradients. This way, the composition of the electro sprayed liquid can be controlled independently of the CZE buffer, thereby providing operation with buffers that could not be used previously, e.g., aqueous and high ionic strength buffers. In addition, the interface operation becomes independent of the CZE flow rate. [62]... 

The most recent modification of the NBD-Cl method involves a further improvement in its qualitative support (616). It involves the infusion of the extract employed for thin-layer chromatography via an electrospray interface into a mass spectrometer operating in the multiple-stage mass spectrometry mode, thus allowing confirmation of suspect results. The cleanup of the thyroid gland samples was also performed with a selective extraction procedure, based on the specific complex formation of the thiouracil, methylthiouracil, propylthiouracil and phenylthiouracil, tapazole, and mercaptobenzimidazole residues with mercury ions bound in a Dowex 1-X2 affinity column. 

Detection of imatinib is performed by triple quadrupole mass spectrometer with an electrospray ionization (ESI) interface operated in positive ion mode [103, 104, 106,107,109,110]. Except the methods published by Parise et al. for imatinib and its main metabolite [108], and for nilotinib [111], where a single quadrupole mass spectrometer was used, most TKIs are analyzed in plasma by atmospheric pressure ionization (electrospray or turbo ion spray) coupled to triple stage mass spectrometer. Expectedly, higher limit of quantifications for imatinib (30 ng/ml) [108], and nilotinib (5 ng/ml) [111], are obtained for the assays using single quadrupole MS (see Table 2). 

Successful operation of the electrospray interface requires the optimization of a variety of interrelated ion-source parameters. In addition, the solvent composition and flow rate may influence the system performance. 

Capillary IC features a flow rate in the range of 10-30 pF/min, thus requiring modifications and reoptimization of existing electrospray interfaces that are usually optimized for either analytical flow (100 pF/min to several milliliters/minute) or nanoflow (<1 pF/min) rates. The optimization of interface parameters such as probe temperature, nebulizer gas, needle voltage, type of desolvation solvent, and the flow rate plays a critical role in establishing instrument sensitivity. When capillary IC is operated at a flow rate of 10-20 pF/min, a probe temperature of 300 °C, a needle voltage of 3 kV, a nebulizer gas pressure of 65 psi, and the use of... 

The total taxane (TTAX) concentration, a sum of the concentrations for CT-2103, TXL, and TXL-metabohtes was determined from scintillation counting of the plasma or tissue sample homogenates. Extractable taxanes, including TXL and organically extractable TXL metabolites, were determined by scintillation counting of ethyl acetate extractions of the plasma, tumour, liver, and spleen samples. Plasma and tissue TXL concentrations were also determined by HPLC/radiometric analysis of the extracts. Metabolites were identified by HPLC followed by mass spectrometry on a Quattro It (Micromass, Manchester, UK) triple quadrupole mass spectrometer fitted with an electrospray orthogonal Z spray ion interface operating in the positive ion mode. ... 

For the 2006—2007 samples, an acetonitrile/water gradient was used at a flow rate of 0.3 mL/min, starting at 5% acetonitrile/95% water/0.05% formic acid, increasing to 40% acetonitrile/60% water/0.05% formic acid in 15 min, and then, finally, to 100% acetonitrile in 10 min, and subsequently held constant for 10 min. Between the runs, the analytical colirrrm was reequihbrated for 15 min. For the 2008—2009 samples, a methanol/water gradient was rrsed at 0.3 mL/min (A) ultrapure water, 0.05% formic acid (B) methanol 0.05% formic acid a linear gradient of 95% A to 0% in 20 min was employed. The mass spectrometer was operated in positive ionization mode irsing an electrospray interface. 

Capillary electrophoresis is usually conducted by applying approximately 30 kV between anode and cathode. In CE/MS with positive-ion operation, the CE anode is at 30 kV, while the CE cathode is at the high voltage of the electrospray interface—for example, at 5 kV. The voltage drop over the column is reduced to 25 kV, resulting in a longer analysis time. 

Thermospray was quite popular before the advent of electrospray, but has now given way to the more robust API techniques, although TSP sources continue to operate. Developed as an LC-MS interface, this technique calls for a continuous flow of sample in solution. 

The ionspray (ISP, or pneumatically assisted electrospray) LC-MS interface offers all the benefits of electrospray ionisation with the additional advantages of accommodating a wide liquid flow range (up to 1 rnl.rnin ) and improved ion current stability [536]. In most LC-MS applications, one aims at introducing the highest possible flow-rate to the interface. While early ESI interfaces show best performance at 5-l() iLrnin, ion-spray interfaces are optimised for flow-rates between 50 and 200 xLmin 1. A gradient capillary HPLC system (320 xm i.d., 3-5 xLmin 1) is ideally suited for direct coupling to an electrospray mass spectrometer [537]. In sample-limited cases, nano-ISP interfaces are applied which can efficiently be operated at sub-p,Lmin 1 flow-rates [538,539]. These flow-rates are directly compatible with micro- and capillary HPLC systems, and with other separation techniques (CE, CEC). 

Many excellent reviews on the development, instrumentation and applications of LC-MS can be found in the literature [560-563]. Niessen [440] has recently reviewed interface technology and application of mass analysers in LC-MS. Column selection and operating conditions for LC-MS have been reviewed [564]. A guide to LC-MS has recently appeared [565]. Voress [535] has described electrospray instrumentation, Niessen [562] reviewed API, and others [566,567] have reviewed LC-PB-MS. For thermospray ionisation in MS, see refs [568,569]. Nielen and Buytenhuys [570] have discussed the potentials of LC-ESI-ToFMS and LC-MALDI-ToFMS. Miniaturisation (reduction of column i.d.) in LC-MS was recently critically evaluated [571]. LC-MS/MS was also reviewed [572]. Various books on LC-MS have appeared [164,433,434,573-575], some dealing specifically with selected ionisation modes, such as CF-FAB-MS [576] or API-MS [577],... 

An electrospray (ES) interface, which also operates at atmospheric... 

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