Improvements in Deuterium Lamps Maximize HPLC ... - T.E.A.M.
Improvements in Deuterium Lamps Maximize HPLC ... - T.E.A.M. Improvements in Deuterium Lamps Maximize HPLC ... - T.E.A.M.
Improvements in Deuterium LampsContinued from page 17Through innovative design and constructionsteps, Agilent DAD andMWD longlife deuterium lamps arealigned precisely during the manufacturingprocess. A computercontrols positions and permanentlysecures the glass bulb of the lampinto the aluminum mounting-ring,thereby creating a pre-alignedproduct for the Agilent 1100 HPLCinstrument. The PerfectFit finalproduct is not only optimized forperformance, but the lamp is easierto handle during both installationand removal.Shine-Through Deuterium Lamps forHigh-Intensity UV-Vis WavelengthsShine-Through Deuterium lamps forthe Agilent 1100 are constructedso that essentially two lamps—oneultraviolet and the other visible—areplaced in series, providing a highintensity broad-spectrum operatingrange of 200 to 800 nm.A Ten-Fold Increase in SensitivityThe overall increase in sensitivitygained when selecting an AgilentShine-Through Deuterium lamp withan Agilent 1100 could be as high astenfold. This is demonstrated inFigure 2, which compares a separationof lysozyme using an AgilentShine-Through Deuterium lamp withthe same separation conductedusing a deuterium lamp not made toAgilent specifications. Whether operatingin the ultraviolet region at 214nm or at 280 nm, the differences arestriking. Both lamps easily detect themain component at approximately13.2 minutes, but only the Agilentlamp on this absorbance scale clearlydetects the low-level impuritiesFigure 2. Lamp Selection Impacts Chromatographic Performance–Separation ofLysozyme and its ImpuritiesImpurity peaks at retention times greater than 16 minutes are clearly present when the instrumentis configured with the Agilent lamp.mAU200150100500-50mAU200150100500-50mAU1512.5107.552.50-2.5mAU1512.5107.552.50-2.5Agilent Shine-ThroughDeuterium Lamp at 214 nm0 2.5 5 7.5 10 12.5 15 17.5 20 min(Time)Non-Agilent Lamp at 214 nm0 2.5 5 7.5 10 12.5 15 17.5 20 min(Time)Agilent Shine-ThroughDeuterium Lamp at 280 nm0 2.5 5 7.5 10 12.5 15 17.5 20 minNon-Agilent Lamp at 280 nm(Time)Clear detectionof low-levelimpurity peaksPoor impurityidentificationClear detectionof low-levelimpurity peaksPoor impurityidentification0 2.5 5 7.5 10 12.5 15 17.5 20 min(Time)13.1513.1713.1513.1718.5718.9218.5718.9220.7120.71O R D E RG U I D EInstrument/Detector Lamp Type Part Number1100 Series DAD/MWD Shine-Through Deuterium 2140-05901100 Series DAD/MWD Long-Life, Shine-Through Deuterium 5181-15301090 Series Diode Array Detector (DAD) Deuterium 79883-600021050 Series Diode Array Detector Deuterium 79883-600021050 Series Multiple Wavelength Deuterium 79883-60002Detector (MWD)18 Separation Times Volume 16 Number 1 2003 www.agilent.com/chem
Improvements in Deuterium LampsContinued from page 18having retention times between16 and 20 minutes in this sample.Expanding the absorbance scale forthis peptide separation at 214 nmclearly shows that the Agilent lampprovides a six- to ten-fold highersignal-to-noise ratio than the non-Agilent lamp (see Figure 3). Byproviding higher sensitivity, Agilentlamps can extend detection capabilitiesand improve quantification attrace levels—for more than 2,000hours of use.Performance TestsWe also tested four lamps, each froma different manufacturer, for alignmenttolerance and noise followingthe ASTM protocol. The followingtable summarizes the results of thesetests. None of the four lamps testedwere correctly aligned for theAgilent 1100 instrument; all hadalignment tolerances greater than100% and noise levels were high—upto 20 times higher than Agilent specificationsfor the wavelengths listedthe table.PerfectFit Agilent LampsAgilent deuterium lamps are differentand better because they arespecifically designed to last longerand to provide enhanced sensitivitywhen using the Agilent 1100 as wellas the 1090 and 1050 Series HPLCline.Figure 3. Agilent Lamps Are Optimized to Maximize Sensitivity–Separationof Lysozyme and its Impurities (expanded scale) at 214 nmPerfectFit Agilent lamps, with increased signal-to-noise, provide increased sensitivity,precision, and accuracy at low levels.mAU605550454035mAU605550454035Agilent Lamp at 214 nm116.5816 17 18 19 20 min(Time)Non-Agilent Lamp at 214 nm116.58Peak 1 S/N = 150Peak 2 S/N = 400Peak 3 S/N = 300Peak 1 S/N = 15Peak 2 S/N = 50Peak 3 S/N = 5016 17 18 19 20 min(Time)For More InformationVisit www.agilent.com/chemand click “Library” then enterpublication 5980-2372E inthe keyword field and click“Search.” Or ask your localAgilent sales representativefor additional information.Improved S/Nensures detectionof all impuritiesNo impuritydetected218.57218.5718.92320.71320.7120.9420.94Performance Tests Find Competitor Lamp Performance Outside SpecificationsAll of these non-Agilent lamps were outside noise level and alignment specifications (>100%).Non-Agilent Lamp Alignment Tolerance (%) ASTM Noise in mAU measured at:Measurement x-dir y-dir z-dir 254 nm 750 nm 254 nm 750 nm 254 nmAgilent Specifications 100% 100% 100%
<strong>Improvements</strong> <strong>in</strong> <strong>Deuterium</strong> <strong>Lamps</strong>Cont<strong>in</strong>ued from page 17Through <strong>in</strong>novative design and constructionsteps, Agilent DAD andMWD longlife deuterium lamps arealigned precisely dur<strong>in</strong>g the manufactur<strong>in</strong>gprocess. A computercontrols positions and permanentlysecures the glass bulb of the lamp<strong>in</strong>to the alum<strong>in</strong>um mount<strong>in</strong>g-r<strong>in</strong>g,thereby creat<strong>in</strong>g a pre-alignedproduct for the Agilent 1100 <strong>HPLC</strong><strong>in</strong>strument. The PerfectFit f<strong>in</strong>alproduct is not only optimized forperformance, but the lamp is easierto handle dur<strong>in</strong>g both <strong>in</strong>stallationand removal.Sh<strong>in</strong>e-Through <strong>Deuterium</strong> <strong>Lamps</strong> forHigh-Intensity UV-Vis WavelengthsSh<strong>in</strong>e-Through <strong>Deuterium</strong> lamps forthe Agilent 1100 are constructedso that essentially two lamps—oneultraviolet and the other visible—areplaced <strong>in</strong> series, provid<strong>in</strong>g a high<strong>in</strong>tensity broad-spectrum operat<strong>in</strong>grange of 200 to 800 nm.A Ten-Fold Increase <strong>in</strong> SensitivityThe overall <strong>in</strong>crease <strong>in</strong> sensitivityga<strong>in</strong>ed when select<strong>in</strong>g an AgilentSh<strong>in</strong>e-Through <strong>Deuterium</strong> lamp withan Agilent 1100 could be as high astenfold. This is demonstrated <strong>in</strong>Figure 2, which compares a separationof lysozyme us<strong>in</strong>g an AgilentSh<strong>in</strong>e-Through <strong>Deuterium</strong> lamp withthe same separation conductedus<strong>in</strong>g a deuterium lamp not made toAgilent specifications. Whether operat<strong>in</strong>g<strong>in</strong> the ultraviolet region at 214nm or at 280 nm, the differences arestrik<strong>in</strong>g. Both lamps easily detect thema<strong>in</strong> component at approximately13.2 m<strong>in</strong>utes, but only the Agilentlamp on this absorbance scale clearlydetects the low-level impuritiesFigure 2. Lamp Selection Impacts Chromatographic Performance–Separation ofLysozyme and its ImpuritiesImpurity peaks at retention times greater than 16 m<strong>in</strong>utes are clearly present when the <strong>in</strong>strumentis configured with the Agilent lamp.mAU200150100500-50mAU200150100500-50mAU1512.5107.552.50-2.5mAU1512.5107.552.50-2.5Agilent Sh<strong>in</strong>e-Through<strong>Deuterium</strong> Lamp at 214 nm0 2.5 5 7.5 10 12.5 15 17.5 20 m<strong>in</strong>(Time)Non-Agilent Lamp at 214 nm0 2.5 5 7.5 10 12.5 15 17.5 20 m<strong>in</strong>(Time)Agilent Sh<strong>in</strong>e-Through<strong>Deuterium</strong> Lamp at 280 nm0 2.5 5 7.5 10 12.5 15 17.5 20 m<strong>in</strong>Non-Agilent Lamp at 280 nm(Time)Clear detectionof low-levelimpurity peaksPoor impurityidentificationClear detectionof low-levelimpurity peaksPoor impurityidentification0 2.5 5 7.5 10 12.5 15 17.5 20 m<strong>in</strong>(Time)13.1513.1713.1513.1718.5718.9218.5718.9220.7120.71O R D E RG U I D EInstrument/Detector Lamp Type Part Number1100 Series DAD/MWD Sh<strong>in</strong>e-Through <strong>Deuterium</strong> 2140-05901100 Series DAD/MWD Long-Life, Sh<strong>in</strong>e-Through <strong>Deuterium</strong> 5181-15301090 Series Diode Array Detector (DAD) <strong>Deuterium</strong> 79883-600021050 Series Diode Array Detector <strong>Deuterium</strong> 79883-600021050 Series Multiple Wavelength <strong>Deuterium</strong> 79883-60002Detector (MWD)18 Separation Times Volume 16 Number 1 2003 www.agilent.com/chem
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