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December 5, 2017, 715004753 Rev. B

Page 82

B.6

Flow cell specifications

The following table lists the flow cell specifications for the 2998 PDA detector. In the ACQUITY Arc
system, the low-dispersion analytical flow cell is standard. In the ACQUITY Arc Bio system, the
inert, low-dispersion analytical flow cell is standard. In an Alliance

®

HPLC system, the analytical

flow cell is standard and the auto-purification, microbore, and semi-preparative are optional flow
cells.

Path length

10 mm (analytical flow cell)

Cell volume

8.4 µL (analytical flow cell)

Pressure limit

6895 kPa (69 bar, 1000 psi)

Wetted materials

• Standard: 316 stainless steel, fluoropolymer, fused silica,

PEEK™

• ACQUITY Arc

®

Bio: Fluoropolymer, fused silica, MP35N,

PEEK, titanium

a. US patent number 5,883,721

Table B–6:

2998 PDA detector flow cell specifications

Description

Volume (



L)

Pathlength
(mm)

Tubing internal
diameter (inches)

Pressure rating
(kPa/bar/psi)

Inlet

Outlet

Analytical

8.4

10.0

0.010

0.010

6895/69/1000

Low-dispersion
analytical

8.4

10.0

0.005

0.005

6895/69/1000

Inert, low-dispersion
analytical

8.4

10.0

0.005

0.005

6895/69/1000

Auto-purification
• analytical
• prep

12.3

0.5

0.010
0.040

0.040

13,790/138/2000

Microbore

2.7

8.0

0.005

0.005

6895/69/1000

Semi-preparative

16.3

3.0

0.020

0.020

6895/69/1000

Cuvette

N/A

10.0

N/A

N/A

N/A

Table B–5:

Optical component specifications (continued)

Attribute

Specification

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Содержание 2998

Страница 1: ...2998 Photodiode Array Detector Overview and Maintenance Guide 715004753 Revision B Copyright Waters Corporation 2015 2017 All rights reserved...

Страница 2: ...December 5 2017 715004753 Rev B Page ii...

Страница 3: ...or the most recent revision of this document consult the Waters website waters com Trademarks Waters Waters Quality Parts THE SCIENCE OF WHAT S POSSIBLE ACQUITY ACQUITY Arc Alliance Empower and MassLy...

Страница 4: ...n all cases where the symbol is used to find out the nature of the potential hazard and any actions which have to be taken Considerations specific to the 2998 Photodiode Array Detector Power cord repl...

Страница 5: ...strument so that it is difficult to disconnect the power cord Equipment misuse notice If equipment is used in a manner not specified by its manufacturer protections against personal injury inherent in...

Страница 6: ...and above normal levels of a compound If sample trays are the same or very similar vary the location of the QC samples in the trays Ensure that QC sample results fall within an acceptable range and e...

Страница 7: ...dustrial Scientific and Medical ISM instrument requirements Group 1 products apply to intentionally generated and or used conductively coupled radio frequency energy that is necessary for the internal...

Страница 8: ...December 5 2017 715004753 Rev B Page viii...

Страница 9: ...i Intended use of the 2998 Photodiode Array Detector vi Calibrating vi Quality control vi EMC considerations vii Canada spectrum management emissions notice vii ISM Classification ISM Group 1 Class B...

Страница 10: ...ices 34 2 7 Plumbing the 2998 PDA detector 38 2 7 1 Connecting the detector to the gas supply 39 2 8 Starting up and shutting down the 2998 PDA detector 40 2 8 1 Starting the detector 40 2 8 2 Monitor...

Страница 11: ...hooting 58 5 Spectral Contrast Theory 61 5 1 Comparing absorbance spectra 61 5 2 Representing spectra as vectors 62 5 2 1 Vectors derived from two wavelengths 62 5 2 2 Vectors derived from multiple wa...

Страница 12: ...tion 83 C 1 1 Clean solvents 83 C 1 2 Solvent quality 83 C 1 3 Preparation checklist 83 C 1 4 Water 83 C 1 5 Use buffers 84 C 1 6 Tetrahydrofuran 84 C 2 Solvent miscibility 84 C 2 1 How to use miscibi...

Страница 13: ...December 5 2017 715004753 Rev B Page xiii C 9 3 Wavelength selection for chromophore detection 91 C 9 4 Mobile phase absorbance 92...

Страница 14: ...December 5 2017 715004753 Rev B Page xiv...

Страница 15: ...le light UV Vis spectrophotometer With a photodiode array of 512 photodiodes and an optical resolution of 1 2 nm pixel the detector operates within a range of 190 to 800 nm The figure below illustrate...

Страница 16: ...a deviation from absorbance versus concentration linearity The absorbance of mobile phase can reduce the linear range Table 1 1 Optics assembly components Component Function Lamp Deuterium source lamp...

Страница 17: ...e entrance of the flow cell and a taper to the internal bore of the flow cell prevents light rays from striking the internal walls of the flow cell The Waters TaperSlit flow cell so called because of...

Страница 18: ...n the array The following figure shows an absorbance spectrum of benzene Note that the wavelength resolution is sufficient to resolve five principal absorption peaks Figure 1 4 Benzene spectrum at dif...

Страница 19: ...electing the appropriate sampling rate A sufficient number of points must fall across a peak to define its shape Thus the definition between peaks is lost at very low sampling rates Empower uses the e...

Страница 20: ...the filter time constant that you select Increasing the filter time constant reduces baseline noise improving the signal to noise ratio Nevertheless increasing the constant too much artificially broad...

Страница 21: ...e effects Greatly decrease baseline noise Shorten and broaden peaks For the highest resolution select an appropriate sampling rate and choose a fast filter time constant For the highest sensitivity se...

Страница 22: ...remains the same 1 5 Computing absorbance data points The absorbance data points are calculated by the detector and then transmitted to the database MassLynx or Empower software The absorbance calcula...

Страница 23: ...t as the absorbance data 1 5 3 Data averaging The data the detector reports to the database Empower or MassLynx can be the average of data points After calculating absorbance the detector averages abs...

Страница 24: ...ingle data point The baseline noise decreases as the number of exposures sample increases Tip The amount of data stored is based on wavelength range spectral resolution runtime and sample rate Specify...

Страница 25: ...s that you select It then subtracts that value from the absorbance value Abs Comp t Abs t CRef t where Abs Comp absorbance compensated Abs absorbance CRef compensation reference t elapsed time after s...

Страница 26: ...December 5 2017 715004753 Rev B Page 26...

Страница 27: ...kit Release notes When unpacking shipping cartons ensure that their contents appear listed on the packing slips and that conversely the items listed on the packing slips are among the contents Recomme...

Страница 28: ...e to allow proper function of the drip management system drain tube to which you can connect a waste reservoir that diverts solvent leaks from the flow cell 2 4 Stacking system modules This procedure...

Страница 29: ...and connected near the system To connect to the electricity source Recommendation When connecting to the electricity source use a line conditioner or an uninterruptible power supply UPS for optimum l...

Страница 30: ...teady green when the lamp is ignited 2 6 Connecting signal cables See also Waters Ethernet Instrument Getting Started Guide The following figure shows the rear panel location of the connectors used to...

Страница 31: ...detector Chart mark Adds a chart mark at 10 of full scale to either or both analog output channels Signal Out 1 and Signal Out 2 and is configurable Auto zero Calculates an offset value that when add...

Страница 32: ...that accompany the instrument control software 2 6 1 1 Single Waters instrument connection In a single Waters instrument system configuration the connection hardware requires only one standard Etherne...

Страница 33: ...multiple Ethernet instruments Network hubs are not supported Figure 2 6 Mixed system of IEEE and Ethernet instruments An inject start trigger is needed for the 2998 PDA detector Figure 2 7 All Etherne...

Страница 34: ...ignal connections between the 2998 PDA detector s rear panel and these devices Waters Alliance Separations Module Waters eSAT IN Module Waters or other manual injector Other manufacturers integrator o...

Страница 35: ...he generation of a stop flow connect cables shown in the following table and figure Requirement To automatically stop the solvent flow within the system in the event of an error condition or hardware...

Страница 36: ...separations module Alliance separations module connector B 2998 PDA detector connector B Pin 4 stop flow red Pin 6 switch 1 red Pin 5 stop flow black Pin 7 switch 1 black Table 2 6 2998 PDA detector...

Страница 37: ...m an injector it performs the corresponding auto zero or inject start function To send an auto zero or chart mark signal from an injector to the detector connect cables as shown in the following table...

Страница 38: ...rs plastic or cloth covered Compression screw assemblies 3 To make plumbing connections to the detector 1 Measure the length of tubing needed to connect the column outlet to the detector inlet Warning...

Страница 39: ...a quarter turn 6 Seat the free end of the column outlet tubing in the fitting of the detector inlet and then tighten the reusable finger tight fitting as in step 5 7 Seat the end of the detector outle...

Страница 40: ...cause the detector to fail the reference energy diagnostic test Ensure that the solvent is correctly composed and that it is of high quality and miscible with any other solvents used in the system Use...

Страница 41: ...10 mL of degassed methanol if the mobile phase does not contain buffers 2 Power off the detector 2 9 Using a cuvette The 2998 PDA detector cuvette option allows for ease of use in Sample handling Warn...

Страница 42: ...using the cuvette when the HPLC instruments are in the idle or static state and operating under identical flow cell conditions 2 9 1 Before you begin Recommendation To ensure accurate results for the...

Страница 43: ...the slot and does not slide forward Ensure that the cuvette does not become dislodged when you are replacing the cuvette holder 4 Gently guide the cuvette holder back into the flow cell assembly until...

Страница 44: ...December 5 2017 715004753 Rev B Page 44...

Страница 45: ...Offices When you contact Waters be prepared to provide this information Nature of the symptom Instrument serial number Solvent s Method parameters sensitivity and wavelength Type and serial number of...

Страница 46: ...lockages in the solvent lines and flow cell Damage to the detector components Microbial growth 3 4 Maintaining the flow cell The flow cell of the 2998 PDA detector requires maintenance when these even...

Страница 47: ...ffers flush the cell with 10 mL of HPLC grade water and then flush with 10 mL of a low surface tension solvent such as methanol Requirement Ensure that the solvent is miscible with the previous mobile...

Страница 48: ...ly s front plate Grasp the handle and gently pull the flow cell assembly toward you 7 Unpack and inspect the new flow cell 8 Square the flow cell assembly in front of the opening and then insert it in...

Страница 49: ...sible and then tighten slightly more using the 1 4 inch flat blade screwdriver 12 Connect the column outlet tubing to the flow cell inlet 13 Connect the waste line to the flow cell outlet 14 Close the...

Страница 50: ...remely hot during operation Warning To avoid eye injury from ultraviolet radiation exposure power off the detector before changing the lamp wear eye protection that filters ultraviolet light keep the...

Страница 51: ...amp dialogue box type the serial number of the new lamp see the label attached to the lamp connector wire and then click OK 3 6 Replacing the fuses The 2998 PDA detector requires two 100 to 240 Vac 50...

Страница 52: ...s of spring loaded fuse holder 3 With minimum pressure withdraw the spring loaded fuse holder Figure 3 6 Removing fuse holder 4 Remove and discard the fuses 5 Ensure that the new fuses are properly ra...

Страница 53: ...cell Before you power on the detector prime the system to fill the flow cell with solvent and remove any air To ensure that the detector is properly aligned and calibrated fill the flow cell with sol...

Страница 54: ...calibration You can also start this procedure manually Notes This procedure can adversely affect spectral library matching and peak purity calibrations You might need to reacquire spectral libraries...

Страница 55: ...oubleshoot Rear panel Result The PDA Detector Rear Panel dialog box appears The following table describes the 2998 PDA detector I O connections Table 4 1 2998 PDA detector analog out event in connecti...

Страница 56: ...correct If after running the diagnostic functions applicable to your problem and troubleshooting the detector you cannot correct an error condition contact Waters Technical Service department 4 2 1 Tr...

Страница 57: ...analysis Table 4 2 Lamp LED indications LED mode and color Description Unlit Indicates that the detector lamp is extinguished Constant green Indicates that the detector lamp is ignited and that the de...

Страница 58: ...ush the flow cell see page 47 or apply slight backpressure of 207 to 345 kPa 2 to 3 bar 30 to 50 psi on the detector waste outlet For example connect a 1 to 2 foot 30 to 60 cm length of 0 009 inch 0 2...

Страница 59: ...2 to 3 bar 30 to 50 psi on the detector waste outlet For example connect a 1 to 2 foot 30 to 60 cm length of 0 009 inch 0 23 mm ID tubing connected to the detector waste outlet Weak lamp Replace the...

Страница 60: ...December 5 2017 715004753 Rev B Page 60...

Страница 61: ...When measured at specific solvent and pH conditions the shape of a compound s absorbance spectrum characterizes the compound The varying extent of UV Vis absorbance occurring at different wavelengths...

Страница 62: ...ce spectral sensitivity set the bench resolution to 1 2 nm Tip Do not include wavelengths where there is no analyte absorbance 5 2 1 Vectors derived from two wavelengths The spectral contrast algorith...

Страница 63: ...versely if any comparison of ratios does not match the corresponding vectors point in different directions 5 3 Spectral contrast angles Spectra of identical shape have vectors that point in the same d...

Страница 64: ...figure the absorbance spectra of two compounds A and B are similar They therefore produce a small spectral contrast angle 3 0 Figure 5 4 Spectra with a small spectral contrast angle Compound A Compou...

Страница 65: ...contrast angle between these absorbance spectra of the same compound is 3 4 Figure 5 5 Normalized absorbance spectra of a compound at two concentrations 5 4 Undesirable effects Shape differences betw...

Страница 66: ...tometric error for all spectral contrast operations the maximum spectral absorbance of a compound should be less than 1 AU Note that the absorbance of the mobile phase reduces the working linear dynam...

Страница 67: ...can help determine whether the shape difference between spectra is significant In general a spectral contrast angle less than its threshold angle indicates that shape differences are attributable to n...

Страница 68: ...December 5 2017 715004753 Rev B Page 68...

Страница 69: ...the failure of individuals to comply with any safety precaution when installing repairing or operating any of its instruments or devices The following symbols warn of risks that can arise when you ope...

Страница 70: ...se precautions when working in the vicinity of such tubing when it is pressurized Wear eye protection Extinguish all nearby flames Do not use tubing that is or has been stressed or kinked Do not expos...

Страница 71: ...ures Warning To avoid personal contamination with biohazards toxic materials or corrosive materials you must understand the hazards associated with their handling Guidelines prescribing the proper use...

Страница 72: ...ktionst chtigkeit verantwortlichen Personen kann zum Entzug der Bedienungsbefugnis des Systems f hren Avvertenza qualsiasi modifica o alterazione apportata a questa unit e non espressamente autorizzat...

Страница 73: ...d k nnen nichtmetallische Schl uche quellen dadurch wird der Berstdruck des Schlauches erheblich reduziert Attenzione fare attenzione quando si utilizzano tubi in materiale polimerico sotto pressione...

Страница 74: ...ion assur e par le mat riel risque d tre d fectueuses Vorsicht Der Benutzer wird darauf aufmerksam gemacht dass bei unsachgem er Verwendung des Ger tes die eingebauten Sicherheitseinrichtungen unter U...

Страница 75: ...ez toujours les fusibles par d autres du type et de la puissance indiqu s sur le panneau proximit du couvercle de la boite fusible de l instrument Vorsicht Zum Schutz gegen Feuer die Sicherungen nur m...

Страница 76: ...icht Zum Schutz gegen Feuer die Sicherungen nur mit Sicherungen ersetzen deren Typ und Nennwert im Abschnitt Sicherungen ersetzen des Kapitels Wartungsverfahren angegeben sind Attenzione per garantire...

Страница 77: ...nd their associated statements can appear on labels affixed to the packaging in which instruments devices and component parts are shipped Safety ground Frame or chassis terminal Fuse Functional ground...

Страница 78: ...December 5 2017 715004753 Rev B Page 78 Upper limit of temperature Lower limit of temperature Temperature limitation Symbol Description...

Страница 79: ...vironmental specifications for the 2998 PDA detector B 3 Electrical specifications The following table lists the electrical specifications for the 2998 PDA detector Table B 1 Physical specifications A...

Страница 80: ...ee 2 A measure of pollution on electrical circuits which may produce a reduction of dielectric strength or surface resistivity Degree 2 refers only to normally nonconductive pollution Occasionally how...

Страница 81: ...254 nm Bandwidth 3 6 nm 3 pixel bunch Warm up time 60 minutes Environmental stability 2 C h Flow cell dry analytical 10 mm Sampling rate 2 points s Linearity b 5 at 2 0 AU propylparaben series at 257...

Страница 82: ...Wetted materials Standard 316 stainless steel fluoropolymer fused silica PEEK ACQUITY Arc Bio Fluoropolymer fused silica MP35N PEEK titanium a US patent number 5 883 721 Table B 6 2998 PDA detector fl...

Страница 83: ...through 0 45 m filters before their use Solvents distilled in glass generally maintain their purity from lot to lot use them to ensure the best possible results C 1 3 Preparation checklist The follow...

Страница 84: ...scibility If you are running a high temperature application consider the effect of the higher temperature on solvent solubility Buffers dissolved in water can precipitate when mixed with organic solve...

Страница 85: ...7 7 15 4 2 Tetrahydrofuran 0 55 66 0 17 220 4 3 Ethyl acetate 0 47 77 1 19 260 4 3 1 propanol 2 30 97 2 15 210 4 3 2 propanol 2 35 117 7 15 4 4 Methyl acetate 0 45 56 3 15 17 260 4 5 Methyl ethyl keto...

Страница 86: ...ibility C 3 Buffered solvents When using a buffer use a good quality reagent and filter it through a 0 45 m filter Do not leave the buffer stored in the system after use Flush all fluid line pathways...

Страница 87: ...lvent degassing Mobile phase difficulties account for 70 or more of all liquid chromatographic problems Using degassed solvents is important especially at wavelengths below 220 nm Degassing provides s...

Страница 88: ...s section describes the solvent degassing techniques that will help you to attain a stable baseline Degassing your solvent also improves reproducibility and pump performance You can use either of the...

Страница 89: ...unit volume of solvent are removed Surface area of the degassing membrane The length of the degassing membrane is fixed in each vacuum chamber To increase the length of membrane you can connect two or...

Страница 90: ...sulfide 290 Toluene 285 Ethylene dichloride 230 Xylene 290 Table C 3 Wavelength cutoffs for different mobile phases Mobile Phase UV Cutoff nm Mobile Phase UV Cutoff nm Acetic acid 1 230 Sodium chlori...

Страница 91: ...to determine the best wavelength for a particular analysis Table C 4 Electronic absorption bands of representative chromophores Chromophore Chemical Configuration max nm max L m cm max nm max L m cm E...

Страница 92: ...centration of the mobile phase affects its absorbance Examples of several mobile phases are given in the table below Tip The absorbances in the table below are based on a 10 mm pathlength Allene C C 4...

Страница 93: ...02 0 01 Triethylamine 1 2 33 2 42 2 50 2 45 2 37 1 96 0 50 0 12 0 04 0 01 Buffers and Salts Ammonium acetate 10 mM 1 88 0 94 0 53 0 29 0 15 0 02 0 01 Ammonium bicarbonate 10 mM 0 41 0 10 0 01 0 01 EDT...

Страница 94: ...rs PIC reagents PIC A 1 vial L 0 67 0 29 0 13 0 05 0 03 0 02 0 02 0 02 0 02 0 01 PIC B6 1 vial L 2 46 2 50 2 42 2 25 1 83 0 63 0 07 0 01 PIC B6 low UV 1 vial L 0 01 0 01 PIC D4 1 vial L 0 03 0 03 0 03...

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