Anal. Chem. 2003, 75, 5978-5983
Incorporation of a Venturi Device in Electrospray Ionization Li Zhou,† Bingfang Yue,† David V. Dearden,† Edgar D. Lee,† Alan L. Rockwood,‡ and Milton L. Lee*,†
Department of Chemistry and Biochemistry, Brigham Young University, Provo, Utah 84602-5700, and ARUP Institute for Clinical and Experimental Pathology, Salt Lake City, Utah 84108
Electrospray ionization has grown to be one of the most commonly used ionization techniques for mass spectrometry, and efforts continue to improve its performance. Typically, the sprayer tip must be very close to the entrance orifice of the mass spectrometer in order to maximize the conduction of ions from the sprayer into the mass spectrometer. However, because of space-charge repulsion, most ions never reach the sampling orifice. In this work, an industrial air amplifier, for which the working mechanism is based on venturi and coanda effects, was added between an electrospray ionization source and a time-of-flight mass spectrometer. When a series of reserpine solutions (0.5, 1.0, 5.0, and 10.0 µM) were monitored using mass spectrometry, an over 5-fold increase in m/z 609.3 ion intensity was measured for a separation distance of 14 mm between the electrospray tip and interface capillary inlet, as compared to when the electrospray tip was in its normal position 1 mm in front of the inlet without the amplifier. When a voltage was applied to the air amplifier to further assist in focusing the electrosprayed ions, an ∼18-fold increase in m/z 609.3 ion intensity was obtained. In addition, a 34-fold reduction in method detection limit was observed. The significance of electrospray ionization (ESI) for mass spectrometry (MS) has recently been emphasized by the awarding of the Nobel Prize to Fenn1 for his original work in this area.2-4 ESI is most recognized today for its application to biomolecules, where high sensitivity is of paramount importance. ESI is known for its high sensitivity; however, it has the potential of becoming even more sensitive. The major limitation in sensitivity with ESI/MS is due to low ion transmission from the ESI source to the sampling orifice and through any stages utilized for transfer of ions from atmosphere to vacuum in the MS. Although the ionization efficiency approaches 100%, the typical ion transmission efficiency from the ESI source to the extraction region of the MS is only 0.01-0.1%.5-7 †
Brigham Young University. ARUP Institute for Clinical and Experimental Pathology. (1) Press Release: The Nobel Prize in Chemistry 2002, the Royal Swedish Academy of Sciences, 2002. (2) Yamashita, M.; Fenn, J. B. J. Phys. Chem. 1984, 88, 4451-4459. (3) Whitehouse, C. M.; Dreyer, R. N.; Yamashita, M.; Fenn, J. B. Anal. Chem. 1985, 57, 675-679. (4) Fenn, J. B.; Mann, M.; Meng, C. K.; Wong, S. F.; Whitehouse, C. M. Science 1989, 246, 64-71. ‡
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During the process of electrospray ionization, analyte ions are generated at atmospheric pressure and transferred into the lowpressure extraction region of the MS via a conductance-limiting aperture located in the high-pressure region.8,9 Gas-phase collisions and Coulombic repulsion that are inevitably involved result in expansion of the ion cloud, directing ions away from the extraction region of the MS, thus decreasing the sensitivity. Although conventional ion optic devices based on Coulombic effects can effectively focus ions in a vacuum, they are largely ineffective in avoiding or reversing ion-cloud expansion generated by gas-phase collisions and Coulombic repulsion at high pressures.10-12 To assist in desolvation and transmission of ions from the electrospray tip to the sampling nozzle for conventional and microbore liquid chromatography (LC) flow rates, Henion et al. reported an “ion spray” device in which a high-velocity sheath flow nebulizing gas was directed past the electrospray tube tip. By optimizing the flow rate for desolvating the electrosprayed ions, an ∼30% increase in ion signal intensity was obtained as compared to a relatively low flow rate. No indication was given as to ion signal improvement compared to a conventional ESI source without the nebulizer-assisted device.13,14 Covey et al.15 developed an ESI source in which a heated gas flow was directed at an angle toward the flow axis of a nebulizerassisted electrospray source and intersected the droplet flow at a region upstream of the orifice of the MS. The intersecting gas flow mixed with the droplet flow in a turbulent fashion and helped to desolvate ions in the droplets and move them toward the orifice. The intersecting flow device reportedly provided an increase in sensitivity of over 10 times and significantly lowered the back(5) Fabbi, J. C.; Rockwood, A. L.; Sin, J. C. H.; Woolley, C.; Lee, E. L.; Jones, J. Proc. 45th ASMS Conf. Mass Spectrom. and Allied Topics; Palm Springs, CA, 1997; p 51. (6) Zook, D. R.; Bruins, A. P. Int. J. Mass Spectrom. Ion Processes 1997, 162, 129-147. (7) Cech, N. B.; Enke, C. G. Mass Spectrom. Rev. 2001, 20, 362-387. (8) Burlingame, A. L.; Boyd, R. K.; Gaskell, S. J. Anal. Chem. 1998, 70, 647R716R. (9) Shaffer, S. A.; Tolmachev, A.; Prior, D. C.; Anderson, G. A.; Udseth, H. R.; Smith, R. D. Anal. Chem. 1999, 71, 2957-2964. (10) Douglas, D. J.; French J. B. J. Am. Soc. Mass Spectrom. 1992, 3, 398-408. (11) Dodonov, A. F.; Kozlovsky, V.; Loboda, A.; Raznikov, V.; Sulimenkov, I.; Tolmachev, A.; Kraft, A.; Wollnik, H. Rapid Commun. Mass Spectrom. 1997, 11, 1649-1656. (12) Javahery, G.; Thomson, B. J. Am. Soc. Mass Spectrom. 1997, 8, 697-702. (13) Henion, J. D.; Covey, T. R.; Bruins, A. P. U.S. Patent 4,861,988, 1989. (14) Mordehai A.; Hopfgartner, G.; Henion, J. D. U.S. Patent 5,352,892, 1994. (15) Covey, T. R.; Anacleto, J. F. U.S. Patent 5,412,208, 1995. 10.1021/ac020786e CCC: $25.00
© 2003 American Chemical Society Published on Web 09/24/2003
ground in the resulting mass spectra. However, it was necessary to carefully control the two flows and the angle between them for stability of the electrospray. Smith et al. improved the sensitivity of ESI by designing a socalled “ion funnel” in the first vacuum stage of the mass spectrometer between the sampling nozzle and skimmer.16-18 This ion funnel consists of a series of cylindrical ring electrodes of progressively smaller internal diameters. By coapplying RF and DC electric fields on the electrode series, the ion cloud is more effectively focused, and the Coulombic-driven ion cloud expansion is reduced under pressures of 10-4 up to 9 Torr. Thus, ions are more efficiently captured, focused, and transmitted as a collimated ion beam from the sampling nozzle orifice to the skimmer. Over 1 order of magnitude increase in ion signal intensity was reported, as compared to a conventional ESI source.19 A recent improvement to this ion funnel is the use of a multicapillary inlet.18 With the combination of multicapillary inlet and ion funnel, Kim et al.16-18 reported ion transmission efficiencies that are 23 times greater than can be obtained with conventional ESI ion optics. However, the ion funnel improves ion transport only at reduced pressures and cannot be applied at atmospheric pressure conditions between the electrospray tip and sampling nozzle, where most ion losses occur. Until now, few papers have reported effective methods of improving ion signals in the high-pressure region between the sprayer and the entrance orifice of the MS. Smith et al.20,21 placed a ring electrode downstream from an ESI sprayer to focus ions into the MS. Franzen22 employed a focusing ring at atmospheric pressure on the inner wall of a heated glass capillary interface for ESI/MS. French et al.23 and Gulcicek et al.24 reported similar designs of a heated silica capillary to assist in desolvation in which end plate and cylindrical lenses were mentioned. These lenses were located at substantial distances from both the sprayer and the inlet orifice of the heated capillary. According to their papers, the electrode rings were useful; however, no mention was made concerning how much they improved ion signal intensities. Finally, Schneider et al.25,26 described an oblong-shaped stainless steel electrode ring that was connected to a high-voltage power supply and placed near the electrospray tip at a potential less than that of the sprayer. It was reported that this lens produced a 2-fold increase in ion signal intensity and a 2-fold reduction in the signal RSD. Other advantages included an increase in formation of multiply charged ions, less critical positioning of the sprayer for (16) Belov, M. E.; Gorshkov, M. V.; Udseth, H. R.; Anderson, G. A.; Smith, R. D. Anal. Chem. 2000, 72, 2271-2279. (17) Kim, T.; Tolmachev, A. V.; Harkewicz, R.; Prior, D. C.; Anderson, G.; Udseth, H. R.; Smith, R. D. Anal. Chem. 2000, 72, 2247-2255. (18) Kim, T.; Udseth, H. R.; Smith, R. D. Anal. Chem. 2000, 72, 5014-5019. (19) Shaffer, S. A.; Tang, K.; Anderson, G. A.; Prior, D. C.; Udseth, H. R.; Smith, R. D. Rapid Commun. Mass Spectrom. 1997, 11, 1813-1817. (20) Olivares, J. A.; Nguyen, N. T.; Yonker, C. R.; Smith, R. D. Anal. Chem. 1987, 59, 1230-1232. (21) Smith, R. D.; Olivares, J. A.; Nguyen, N. T.; Udseth, H. R. Anal. Chem. 1988, 60, 436-441. (22) Franzen, J. B. U.S. Patent 5,747,799, 1998. (23) French, J. B.; Reid, N. M.; Buckley, J. A. U.S. Patent 4,121,099, 1978. (24) Gulcicek, E. E.; Whitehouse, C. M. U.S. Patent 5,432,343, 1995. (25) Schneider, B. B.; Douglas, D. J.; Chen, D. D. Y. Rapid Commun. Mass Spectrom. 2001, 15, 2168-2175. (26) Schneider, B. B.; Douglas, D. J.; Chen, D. D. Y. J. Am. Soc. Mass Spectrom. 2002, 13, 906-913.
Figure 1. Schematic diagram of the ESI/air amplifier/TOFMS system: (1) ESI source, (2) air amplifier entrance, (3) air amplifier exit, (4) metallic heated capillary (capillary inlet), and (5) nozzle.
Figure 2. Schematic diagram of the air amplifier: (1) nitrogen supply inlet, (2) annular chamber, (3) annular gap, (4) induced airflow, and (5) output air flow.
optimum performance, and more ease in use compared to the ion funnel and intersecting flow devices. An alternative to focusing the electrospray ion beam toward the sampling nozzle is to place the electrospray tip as close to the nozzle as possible so that a large portion of the spray enters the vacuum region. Low flow rates from small-bore ESI tips are desirable for stability of the “Taylor cone” and production of fine electrospray droplets. This combination has been accomplished using microspray and, especially, nanospray sources. The improvement in response can be explained by the fact that sprayed droplets are already small enough to produce gas-phase ions directly. Analyte concentrations down to 1 fmol can be easily sprayed without sheath flow or pneumatic assistance for MS detection.7,27 The closeness of the electrospray tip to the sampling orifice is limited by the electrical discharge threshold between the highvoltage sprayer and the nozzle counter electrode, which is dependent on the voltage applied to the sprayer tip. To overcome these limitations, Jorgenson et al.28 and Sheehan et al.29,30 reported low-pressure electrospray devices, in which analyte solutions were electrosprayed inside the vacuum chamber at reduced pressures. When the electrospray device was positioned in a very low pressure region, Plazer et al.32 reported significant loss of analytes (27) Karas, M.; Bahr, U.; Dulcks, T. Fresenius’ J. Anal. Chem. 2000, 366, 669676. (28) Jorgenson, J. W.; Dohmeier, D. M. U.S. Patent 5,115,131, 1992. (29) Sheehan, E. W. U.S. Patent 5,838,002, 1998. (30) Sheehan, E. W.; Willoughby, R. C.; Jarrell, J. A.; Strand, D. M. U.S. Patent 6,278,111, 2001. (31) Foret, F.; Zhou, H.; Gangi, E.; Karger, B. L. Electrophoresis 2000, 21, 13631371.
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Figure 3. Mass spectra of a 1.0 µM reserpine solution obtained using an ESI/TOFMS and a separation distance of 14 mm between the ESI tip and capillary inlet: (A) without air amplifier; (B) with air amplifier and applied voltage, but no gas flow; (C) with air amplifier and gas flow, but no applied voltage; and (D) with air amplifier, gas flow, and applied voltage. Conditions: 5 kHz pulser frequency, 1 min run time, direct infusion at 1.5 µL min-1, RSD ) (20%.
and appearance of fine droplets on the walls of the vacuum chamber and heated transfer line, thus resulting in a serious decrease in sensitivity. On the other hand, Foret et al.31 reported an improvement in ion signal intensity for CE/MS using a liquidjunction ESI interface at slightly subatmospheric pressure compared to using a liquid sheath ESI interface. It is generally believed that ESI technology has reached the point where modifications produce only minor gains in ion transmission in atmospheric pressure regions of ESI sources.19 New approaches are sorely needed to take advantage of the high sensitivity that is potentially available. In this vein of thought, we report the use of a commercial air amplifier to assist in focusing and desolvating ions in ESI. This device generates a concentric high-velocity converging gas flow around the electrospray tip to reduce spreading of the electrospray plume, to assist in desolvation, and to improve conduction of ions to the sampling orifice of the MS. Applying a voltage to the air amplifier assists in focusing and conduction of ions. EXPERIMENTAL SECTION Materials. Reserpine was purchased from Sigma (St. Louis, MO). HPLC grade methanol, water, and glacial acetic acid were obtained from Mallinckrodt Baker (Paris, KY). Reserpine solutions were prepared by dissolving the salt in a mixture of methanol/ water/acetic acid (69.5:29.5:1 v/v) at concentrations of 0.5, 1.0, 5.0, and 10.0 µM, respectively. Instrumentation. Microspray configurations were utilized for low-flow ESI (1.0-1.5 µL min-1). Microspray tips were prepared (32) Platzer, B. WO Patent 9534089, 1995.
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by cutting 90-µm-o.d., 20-µm-i.d. fused-silica capillary tubing (Polymicro Technologies, Phoenix, AZ) into ∼1.0 in. lengths, which promoted the onset of electrospray at relatively low voltages.33 A Jaguar time-of-flight mass spectrometer (TOFMS, LECO, St. Joseph, MI) with a homemade heated capillary interface (Figure 1) was used to test the air amplifier focusing system. An aluminum industrial air amplifier (HMC-Brauer, Mount Farm, Milton Keynes, U.K.) was remachined out of stainless steel and placed between the sprayer tip and interface capillary.34 Two high-voltage power supplies (series 230, Bertan, Hicksville, NY) were connected to the ESI source and air amplifier, respectively. The voltages on the ESI source, air amplifier, interface capillary, and skimmer were set at +2.8 to +4.0 kV, 0.0 to +3.0 kV, +300 V, and +65 V, respectively. The air amplifier was grounded, except when a voltage was applied. The various reserpine solutions were introduced at an infusion flow rate of 1.5 µL min-1. RESULTS AND DISCUSSION A schematic diagram of the air amplifier is shown in Figure 2. High-velocity compressed nitrogen gas at 4-6 L min-1 flows from the supply inlet (1) and into the annular chamber (2). The flow is throttled by the annular gap (3), and the resultant thin layer of high-velocity nitrogen gas adheres to the wall profile, which turns the nitrogen gas through 90° to pass down the bore. The action (33) Lazar, I. M.; Lee, E. D.; Sin, J. C. H.; Rockwood, A. L.; Onuska, K. D.; Lee, M. L. Am. Lab. 2000, 2, 110-119. (34) Chu, I. H.; Zhang, H.; Leming, S.; Dearden, D. V. J. Am. Chem. Soc. 1993, 115, 5736-5744.
Table 1. Amplification Factors for Reserpine m/z 609.3 Ion Intensity Obtained Using an ESI/Air Amplifier/TOFMS base peak intensity/amplification factor (IB/(IA - 1)) with air amplifierb(IB) concn (µM)
without air amplifiera (IA)
applied voltage, no gas flowc
gas flow, no applied voltaged
applied voltage, gas flowe
0.5 1.0 5.0 10.0
134/223/1111/3394/-
228/0.7 357/0.6 1778/0.6 5091/0.5
1131/7.4 1727/6.7 7812/6.0 21689/5.4
2809/20.0 4311/18.3 26592/22.9 63480/17.7
a Signal obtained when the electrospray tip was positioned 1 mm from the sampling orifice of the TOFMS; RSD ) (16%. b Signal obtained when the electrospray tip was positioned 14 mm from the sampling orifice of the TOFMS. c RSD ) (19%. d RSD ) (16%. e RSD ) (20%.
Figure 4. Reserpine mass spectral base peak intensity (m/z 609.3) versus on-axis distance between the ESI tip and capillary inlet: (A) with air amplifier, capillary inlet fixed at 25.5 mm inside the exit of the air amplifier, and the ESI tip moved along the axis; (B) with air amplifier, ESI tip fixed at 6 mm inside the entrance of the air amplifier, and the capillary inlet position changed along the axis; and (C) without the air amplifier. Conditions: 5 kHz pulser frequency; 1 min run time; direct infusion at 1.5 µL min-1; RSD ) (17, 14, and 21% for A, B, and C, respectively.
of the high-velocity nitrogen gas streaming down the bore of the air amplifier causes a pressure drop, which induces a large flow of ambient air into the device (4). The net effect is that the venturi device uses the energy from a small volume of compressed nitrogen gas to produce a large volume, large velocity, and lowpressure outlet air flow (5). The outlet flow volume can be as high as 100 times the supply flow, that is, 400-600 L min-1. To test the air amplifier focusing system, ion signal enhancements were studied using the instrumental setup shown in Figure 1. A series of reserpine concentrations were analyzed under the conditions of (1) no air amplifier; (2) with air amplifier and applied
voltage (1.9-2.0 kV), but no venturi-induced gas flow; (3) with air amplifier and venturi-induced gas flow, but no applied voltage; and (4) with air amplifier, venturi-induced gas flow, and applied voltage. The peak intensities for m/z 609.3 were monitored. Ten determinations of each measurement were made for statistical considerations. The capillary interface was heated to 75 °C. Figure 3 shows examples of mass spectra obtained for 1.0 µM reserpine solution using the ESI-TOFMS without air amplifier (Figure 3A); ESI/air amplifier without gas flow, but with applied voltage (Figure 3B); ESI/air amplifier without applied voltage, but with gas flow (Figure 3C); and ESI/air amplifier with both gas flow and applied voltage (Figure 3D). These experiments were performed with the ESI tip axially positioned 6 mm inside the entrance of the air amplifier and the capillary inlet axially positioned 22.5 mm inside the exit of the air amplifier (i.e., the ESI tip was positioned 14 mm from the capillary inlet along the axial direction). The greatest enhancement in ion signal intensity was observed when gas flow and voltage were used together with the air amplifier. With no air flow and 1.9 to 2.0 kV applied to the air amplifier (4.0 kV ESI voltage), the m/z 609.3 ion signal intensity increased by over 50%, as compared to when the electrospray tip was positioned 1 mm in front of the capillary inlet (i.e., no air amplifier) and 2.8 kV was applied to the ESI source (Figure 4C). With gas flow through the air amplifier and no voltage applied, over a 5-fold increase (amplification factor) was obtained. When 4.0 kV was applied to the ESI source and 1.9-2.0 kV was applied to the air amplifier with venturi-induced gas flow, we obtained an ∼18-fold increase (amplification factor) in ion signal intensity (Table 1). To find the optimum positions of the ESI tip and capillary inlet as described above, the ESI tip, air amplifier, and capillary inlet positions were axially changed relative to each other until the measured ion intensity was at a maximum. This was accomplished by moving the ESI tip from 12 mm inside the entrance of the air amplifier to 20 mm outside the entrance of the air amplifier at 1 mm increments and, at each increment, moving the ESI tip and air amplifier axially together so that the capillary inlet was axially positioned from 25.5 mm inside the exit of the air amplifier to 8.5 mm outside the exit of the air amplifier. It was found that when the electrospray tip was axially positioned 6 mm inside the Analytical Chemistry, Vol. 75, No. 21, November 1, 2003
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Figure 5. Reserpine mass spectral base peak intensity (m/z 609.3) versus off-axis distance for (A) ESI tip, with air amplifier, and the capillary inlet axially fixed; and (B) capillary inlet, with air amplifier, and the ESI tip axially fixed. Conditions: 5 kHz pulser frequency; 1 min run time; direct infusion at 1.5 µL min-1; RSD ) (21 and 14% for A and B, respectively. Table 2. Method Detection Limits Using an ESI/Air Amplifier/TOFMS
condition(s) no air amplifier air amplifier with gas flow air amplifier with gas flow and 1.9-2.0 kV applied
Figure 6. Reserpine mass spectral base peak intensity (m/z 609.3) versus reserpine concentration (logarithmic scale) for (A) with air amplifier, gas flow, and applied voltage; (B) with air amplifier and gas flow, but no applied voltage; and (C) without the air amplifier. Conditions: 5 kHz pulser frequency, 1 min run time, direct infusion at 1.5 µL min-1, RSD ) (20%. The dotted line represents the experimental noise level.
entrance of the air amplifier and the capillary inlet was axially positioned 22.5 mm inside the exit of the air amplifier (i.e., the ESI tip was positioned 14 mm from the capillary inlet along the axial direction), the ion intensity reached its peak value. The results of two of these experiments are shown in Figure 4A and B. Figure 4A shows the base peak intensity as a function of distance between the ESI tip and capillary inlet when the capillary inlet was axially fixed 22.5 mm inside the exit of the air amplifier. Figure 4B shows the base peak intensity as a function of distance between the ESI tip and the capillary inlet when the ESI tip was axially fixed 6 mm inside the entrance of the air amplifier. The highest ion intensity was observed when the capillary inlet was positioned 22.5 mm inside the exit of the air amplifier and the ESI tip was positioned 6 mm inside the entrance of the air amplifier, that is, the ESI tip was positioned 14 mm from the capillary inlet along the axial direction. Furthermore, a relatively broad range of ESI tip and capillary inlet positions was found for maintaining strong ion signal intensities. Even when the distance between the ESI tip and capillary inlet was 20 mm, the ion intensity was still higher than when the ESI tip was 5982 Analytical Chemistry, Vol. 75, No. 21, November 1, 2003
detection limits (fmol)
detection limits (nmol/L)
reduction (fold)
5.30 0.64 0.16
3.53 0.43 0.11
8.3 33.9
positioned 1 mm in front of the sampling orifice without the air amplifier (Figure 4C). To evaluate the relationship between ion intensity and off-axis distance of the ESI tip or the capillary inlet from their optimum positions, each was moved off-axis while the other was axially fixed in its optimum position. When the ESI tip was moved off-axis by (2 mm while the capillary inlet was axially fixed, the ion intensity decreased by 40% (Figure 5A). When the capillary inlet was positioned off-axis by (2 mm while the ESI tip was axially fixed, the ion intensity decreased by 19% (Figure 5B). Very little loss in ion signal intensity was observed when the electrospray tip or the capillary inlet was moved (1 mm off-axis. Finally, the base peak intensity (m/z 609.3) was plotted against concentration with the air amplifier in its optimum position (Figure 6). After linear regression, the method detection limits were calculated on the basis of concentrations corresponding to three times the signal-to-noise ratio. A 34-fold reduction in method detection limit was obtained (Table 2). In addition to enhancing analyte ion intensity, the air amplifier also suppresses background chemical noise. The gain in ion signal intensity is attributed to the ability of the air amplifier to stabilize the electrospray and improve conduction of ions into the MS. The ESI tip can be located farther from the sampling orifice than for conventional ESI to produce better desolvation and less possibility of discharge. Another advantage of the device is that the sprayer can be positioned along the axial direction straight toward the nozzle. Complex devices with offaxis orientation of the ESI needle with respect to the nozzle for separating ions from neutrals and improving desolvation are not necessary.35
CONCLUSIONS A commercial air amplifier assisted in focusing electrosprayed ions along the axial direction toward the sampling orifice of a mass spectrometer. Using an ESI/TOFMS, a signal enhancement of ∼18 times and a detection limit reduction of ∼34 times were
observed when using the air amplifier, as compared to conventional micro ESI.
(35) Taflin, D. C.; Ward, T. L.; Davis, E. J. Langmuir 1989, 5, 376-384.
AC020786E
Received for review December 30, 2002. Accepted August 1, 2003.
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