Electrospray efficiency

Hence, as previously mentioned, interfacing chiral CE with ESI/MS is severely hampered by the presence of nonvolatile additives, such as CD, which often leads to a significant loss of electrospray efficiency and ion source contamination. In order to circumvent these limitations, several approaches have been investigated, such as fhe combined column coupling with voltage switching and the partial-filling technique. 

The Z-spray inlet/ion source is a particularly efficient adaptation of the normal in-line electrospray source and gets its name from the approximate shape of the trajectory taken by the ions between their formation and their entrance into the analyzer region of the mass spectrometer. A Z-spray source requires much less maintenance downtime for cleaning. 

True electrospray is most efficient at flow rates of between 5 and 10 p,lmin , which are not directly compatible with the majority of HPLC applications. There are two approaches to providing reduced flow rates of an appropriate magnitude. 

The effect of the buffer on the efficiency of electrospray ionization was mentioned earlier in Section 4.7.1. This is a good example of the dramatic effect that this may have - good chromatographic separation and ionization efficiency with formic, acetic and propionic acids, and good separation, although with complete suppression of ionization, with trifluoroacetic acid (TFA), the additive used for the protein application described previously. Post-column addition of propionic acid to the mobile phase containing TFA has been shown to reduce, or even... 

The applicable HPLC flow rate with ESI is lower than that with thermospray or APCI, usually below the O.SmLmin range. The typical flow rate is 0.10-0.20 mL min for ESI, which means that the effluent flow introduced into the electrospray must be reduced by splitting when using a conventional HPLC column (4.6-mm i.d. x 250 mm). Currently, narrower columns (e.g., 2.1-mm i.d.) and slower flow rates are commonly used to achieve the desirable flow rates. The advantage of this approach is that improved separation efficiency and faster separations are also achieved (at the cost of sample capacity). 

ES ionisation can be pneumatically assisted by a nebulising gas a variant called ionspray (IS) [129]. ESI is conducted at near ambient temperature too high a temperature will cause the solvent to start evaporating before it reaches the tip of the capillary, causing decomposition of the analyte during ionisation and too low a temperature will allow excess solvent to accumulate in the sources. Table 6.20 indicates the electrospray ionisation efficiency for various solvents. 

Table 6.20 Electrospray ionisation efficiency Solution in water ...

As the vast majority of LC separations are carried out by means of gradient-elution RPLC, solvent-elimination RPLC-FUR interfaces suitable for the elimination of aqueous eluent contents are of considerable use. RPLC-FTTR systems based on TSP, PB and ultrasonic nebulisa-tion can handle relatively high flows of aqueous eluents (0.3-1 ml.min 1) and allow the use of conventional-size LC. However, due to diffuse spray characteristics and poor efficiency of analyte transfer to the substrate, their applicability is limited, with moderate (100 ng) to unfavourable (l-10pg) identification limits (mass injected). Better results (0.5-5 ng injected) are obtained with pneumatic and electrospray nebulisers, especially in combination with ZnSe substrates. Pneumatic LC-FI1R interfaces combine rapid solvent elimination with a relatively narrow spray. This allows deposition of analytes in narrow spots, so that FUR transmission microscopy achieves mass sensitivities in the low- or even sub-ng range. The flow-rates that can be handled directly by these systems are 2-50 pLmin-1, which means that micro- or narrow-bore LC (i.d. 0.2-1 mm) has to be applied. 

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). 

Yiang, Y. Hofstadler, S. A. A highly efficient and automated method of purifying and desalting PCR products by electrospray ionization mass spectrometry. Anal. Biochem. 2003,316, 50-57. 

Although El MS is an efficient way to provide structural information on several molecular constituents of various lipid substances it only provides partial information and it is particularly not suitable for the study of the low volatile components. High molecular weight and nonvolatile compounds are particularly difficult to analyse in this way and it may therefore be interesting to explore the possibilities of other ionisation modes such as electrospray for an accurate structural study of high molecular constituents such as monoester and diester species of beeswax (Gamier et al., 2002) and TAGs of animal fats... 

One reason for lower sensitivity is the lack of flexibility to optimize the positions of the sprayers on the MUX interface another may be the lower electrospray desolvation efficiency on the MUX. The longer total cycle time on a MUX interface with a quadrupole MS in comparison to a single sprayer interface adds another concern. Assuming typical chromatographic peak widths appeared on average at 15 sec, 17 data points could be easily detected across the peak for each transition with a total cycle time of 0.88 sec on a conventional single sprayer set-up. With the MUX, only 12 data points could be detected across the same peak even with a total cycle time of 1.24 sec because of the introduction of additional interspray time on top of dwell time. Hence, when MUX is used with a quadrupole mass analyzer, it is important to consider dwell time and chromatographic peak width... 

Megaflow electrospray An electrospray system capable of producing droplets directly from HPLC flow rates of the order of 1 ml min-1 (true electrospray is most efficient at flow rates of the order of 10 p.lmin-1). 

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