Forced degradation analysis of recombinant adeno-associated virus serotype 8 based on analytical anion exchange chromatography coupled to orthogonal characterization

Oligonucleotide and mRNA Analysis Resource Roundup

NIST26 introduces integrated chromatogram processing, deconvolution, and library searching for EI GC-MS and LC-MS/MS data, but the workflow begins with one essential requirement: chromatographic data must be available in the mzML format. In his presentation <a target="_blank" rel="noopener noreferrer" href="https://labrulez.com/pdf/Conversion_to_mz_ML_63e134bb34.pdf">Conversion to mzML Format for NIST26 Chromatogram Processing</a>, James Little of Mass Spec Interpretation Services explains how vendor data can be converted to mzML using either instrument-vendor software or the freely available MSConvert tool from ProteoWizard, and highlights several settings that are critical for successful processing in NIST26.<h4>NIST26 Chromatogram processing requires mzML</h4>For the chromatogram-processing workflow described in the presentation, mzML is the accepted input format. Proprietary LC-MS or GC-MS data therefore need to be converted before they can be loaded into the NIST26 chromatogram-processing environment.There are two principal ways to obtain an mzML file:<ul><li>conversion using software supplied by the instrument manufacturer,</li><li>conversion using MSConvert, which is included in the free ProteoWizard software package.</li></ul>Using an open format such as mzML also provides a practical way to move mass spectrometry data between different software environments without relying exclusively on proprietary vendor formats.<h4>Using ProteoWizard and MSConvert</h4>MSConvert is installed as part of the complete ProteoWizard package rather than as a separate application. After installing ProteoWizard, users can access either the graphical MSConvert interface or the command-line version.The presentation recommends the stable release for routine work, while noting that newer or nightly builds may sometimes provide improved support for recently introduced vendor formats or additional control over data-processing filters. This can be particularly relevant when handling vendor files in which centroiding behavior or peak picking requires closer attention.For most users, the graphical interface provides a straightforward route from proprietary raw data to an mzML file suitable for further NIST26 processing.<h4>Start by selecting mzML as the output format</h4>The first step in MSConvert is to select the source file or dataset using the Browse function.The software supports a broad range of mass spectrometry data formats, and the file-selection window can either be restricted to a particular vendor format or set to display any supported spectral format.After selecting the source data, the output format must be set to mzML. The screenshots in the presentation show this setting directly in the MSConvert options panel before the conversion is started.The user can also select the destination folder and other file-output options before adding the selected dataset to the conversion queue.<h4>Add the data and start the conversion</h4>Once the source file and output settings have been selected, the file is added to the conversion list using the Add command.The conversion itself is then launched with Start.For simple datasets that are already stored in the correct spectral representation, this may be all that is required. However, James Little emphasizes an additional requirement that is particularly important for NIST26: the data must be centroided.<h4>Centroided data are required</h4>According to the presentation, profile-mode data are not acceptable for this NIST26 chromatogram-processing workflow. The spectra must instead be in centroided, or peak-picked, form.Profile spectra contain the full measured shape of each mass spectral signal across the m/z axis. Centroided data reduce these profiles to discrete peaks represented by their characteristic m/z and intensity values.If the original vendor data are still in profile mode, centroiding can be performed during conversion by adding the Peak Picking filter in MSConvert.This means that the conversion step can perform two operations at once:vendor raw data → centroiding / peak picking → mzMLThe resulting mzML file can then be used as input for NIST26 chromatogram processing.<h4>Peak Picking must be the first filter</h4>One of the most important practical details in the presentation is the position of the Peak Picking operation in the MSConvert filter sequence.If centroiding is required, Peak Picking must be the first algorithm or filter in the list. Placing another processing function before it can interfere with the intended conversion workflow.MSConvert allows the user to select the Peak Picking filter, define the relevant MS levels, and add it to the processing sequence before starting conversion. The example shown in the presentation applies vendor-based peak picking to selected MS levels.If there is uncertainty about whether the source data are already centroided, the Peak Picking filter can still be included. According to the presentation, applying it to data that are already centroided does not alter the file unnecessarily.This provides a useful safeguard when the acquisition mode of an older dataset is uncertain.<h4>Selecting the appropriate MS levels</h4>Peak picking does not necessarily have to be applied identically to every spectrum in a dataset. MSConvert allows users to specify the MS levels to which the filter should be applied.This is especially relevant for LC-MS/MS data, where a file can contain both precursor-level MS spectra and tandem MS spectra.The conversion settings should therefore reflect the way the original data were acquired and how those data will subsequently be processed in NIST26.<h4>Be aware of changes in file size</h4>The presentation also notes that converted files may become substantially different in size from the original vendor files.This is worth considering when working with large LC-MS/MS datasets or processing batches of chromatograms. The size of the resulting mzML file can depend on the original acquisition mode, centroiding, compression settings, the number of MS levels, and other output parameters.Storage requirements should therefore be considered before converting large collections of raw data.<h4>A simple workflow for preparing data for NIST26</h4>The overall procedure presented by James Little can be summarized as a short sequence:<ol><li>Obtain the original vendor LC-MS or GC-MS data.</li><li>Open the data in MSConvert or use the vendor's own conversion software.</li><li>Select mzML as the output format.</li><li>Check whether the spectra are centroided.</li><li>If necessary, add Peak Picking as the first processing filter.</li><li>Select the appropriate MS levels.</li><li>Add the file to the conversion list.</li><li>Start the conversion.</li><li>Use the resulting mzML file for NIST26 chromatogram processing.</li></ol>The procedure itself is straightforward, but the centroiding requirement is important. A file can have the correct <code>.mzML</code> extension and still be unsuitable for the workflow if its spectra remain in profile mode.<h4>From vendor data to integrated NIST26 processing</h4>File conversion may appear to be a relatively minor step compared with deconvolution, spectral interpretation, and library searching, but it establishes the foundation for the entire downstream workflow.By using mzML as the common input format, NIST26 can process data originating from different instrument platforms through a more consistent workflow. ProteoWizard and MSConvert provide a practical bridge between proprietary mass spectrometry files and this standardized format.The key message from the presentation is therefore simple: convert to mzML, ensure the spectra are centroided, and place Peak Picking first whenever centroiding is required. Once those conditions are met, the data are ready for the integrated chromatogram-processing and library-searching capabilities available in NIST26.

Conversion to mzML Format for NIST26 Chromatogram Processing

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Waters/Wyatt ViscoStar HPLC viscometer

Waters ViscoStar is a highly sensitive, on-line differential viscometer used in conjunction with SEC-MALS-IV to determine the size and conformation of all types of biopolymers, synthetic polymers and even proteins and peptides.

<h4>Highly Sensitive</h4>A highly sensitive, on-line differential viscometer used in conjunction with SEC-MALS-IV to determine the size and conformation of all types of biopolymers, synthetic polymers and even proteins and peptides.<h4>Multiple Novel Technologies</h4>The ViscoStar incorporates multiple novel technologies to provide the highest sensitivity, stability and solvent compatibility of any available viscometer for GPC. Its ease-of-use and serviceability make it the perfect companion for Wyatt's DAWN™ light scattering and Optilab™ refractive index detectors.<h4>Another path to polymer and protein characterization: SEC-MALS-IV</h4>When coupled with size-exclusion chromatography and a concentration detector such as the Optilab™ , the ViscoStar derives the intrinsic viscosity (IV) of polymers. An inherently valuable physico-chemical property in itself, IV is best utilized together with molar mass information from multi-angle light scattering (SEC-MALS-IV) to derive the hydrodynamic radius, conformation, and branching ratio of a macromolecule in solution.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Visco_Star_pump_pulse_suppression_with_inset_e8c2fbfc18.jpg" alt="Waters ViscoStar: Patented pulse compensation technology in the ViscoStar eliminates pump fluctuations. At the hardware level, the bridge is impedance matched for dynamic balance, greatly reducing the effect of pump pulses. An additional layer of software filtering eliminates residual pump pulses." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Visco_Star_pump_pulse_suppression_with_inset_e8c2fbfc18.jpg 233w," sizes="100vw" width="348" height="233"><h5>Key benefits of the ViscoStar</h5><ul><li>High sensitivity, equivalent to 100 ng of 100 kDa polystyrene in THF at 1.0 mL/min injected onto a standard GPC column</li><li>High dynamic range of over 135,000:1. The full measurement range corresponds to over 13 mg/mL of 100 kDa polystyrene in THF</li><li>Extended solvent compatibility with all-316 stainless steel transducers</li><li>Temperature range from 4 °C to 70 °C for polymers or proteins that require a range of temperatures</li><li>Minimal band broadening thanks to fast pressure transducers</li><li>Pump pulse suppression to below 4 Pa (hardware alone) or 1 Pa (hardware + software) without loss of chromatographic resolution</li><li>Automated capillary bridge tuning with no moving parts, to guarantee a perfectly balanced bridge, every run</li><li>Enhanced thermal stability with drift of &lt;2.5 Pa/hour</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Visco_Star_cmc_plga_intrinsic_viscosity_graphs_be99c5d115.jpg" alt="Waters ViscoStar: The ViscoStar provides the highest available dynamic range and sensitivity in online viscometry. This graph shows the intrinsic viscosities of 1) poly(lactic co-glycolic acid), or PLGA, in THF and 2) carboxymethyl cellulose, or CMC, in aqueous mobile phase, both measured with a ViscoStar and Optilab, spanning a range from tens to thousands of mL/g. The same instruments readily measure intrinsic viscosity values in the low single digits with excellent signal-to-noise, even at very low concentrations." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Visco_Star_cmc_plga_intrinsic_viscosity_graphs_be99c5d115.jpg 236w," sizes="100vw" width="406" height="268"><h5>SEC-IV without MALS?</h5>Yes, as much as we love MALS for optimal polymer analysis, it's true that there are occasions when it just won't fit the bill.Universal calibration is a parallel technique to SEC-MALS for determining molecular weight, commonly used to characterize linear polymers. While universal calibration overcomes some of the problematic issues associated with column calibration by reference molecules, it does not account for non-ideal column behavior caused by chemical solute-column interactions or hyperbranching.Universal calibration is particularly-well suited to replace MALS for:<ul><li>Strongly fluorescent polymers, in particular those not adequately addressed by a DAWN™ with long-wavelength (785 nm) laser or a standard DAWN (685 nm) with fluorescence-blocking filters.</li><li>Weakly-scattering polymer/solvent systems with very low dn/dc values</li></ul>The Mark-Houwink-Sakurada relationship makes use of empirical relationships between intrinsic viscosity and molar mass, specific to each polymer and solvent, to estimate molar mass from SEC-IV measurements. Of course, the best way to determine MHS coefficients is to measure them using SEC-MALS-IV and ASTRA™!<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Visco_Star_v3_protein_rh_measurements_0951d3769a.jpg" alt="Waters ViscoStar: Three IgG proteins with similar MWs eluted at different elution volumes. To understand their different elution properties, online DLS is typically used to measure the hydrodynamic radius, R h , of a protein. However, online DLS did not provide reliable insights, in this case due to the relatively high experimental uncertainty of the DLS. The precise Rh values obtained with a MALS detector and the ViscoStar III, on the other hand, clearly revealed the causes. IgG B eluted later than IgG A due to its more compact structure; IgG C had a slight interaction with the SEC column." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Visco_Star_v3_protein_rh_measurements_0951d3769a.jpg 237w," sizes="100vw" width="401" height="264"><h4>Specifications</h4><h5>Measurement Capabilities – Differential Viscosity</h5><ul><li>Sensitivity: Detects as little as 0.1 µg of 100 kDa polystyrene in THF at 1 mL/min under typical GPC conditions.</li><li>Differential Pressure Performance:<ul><li>Noise: &lt; 0.05 Pa</li><li>Range: 6.9 kPa, with built-in overpressure protection</li><li>Drift: &lt; 2.5 Pa/hr</li><li>Linearity: Within 0.5% of full scale</li><li>Dynamic Range: 135,000:1</li></ul></li><li>Capillary Bridge Autobalance: Thermally regulated, leak-free mechanism ensures stable baseline and precision.</li><li>Sample Shear Rate: Up to 5000 Hz with water at 1 mL/min</li></ul><h5>Fluidics and Flow Compatibility</h5><ul><li>Maximum Flow Rate:<ul><li>Aqueous: 1.5 mL/min</li><li>THF: 3 mL/min</li></ul></li><li>Delay Volume Options: Configurable at 16.2, 10.8, or 5.4 mL; optional factory-installed upgrade to 32.4 mL (Note: actual column fluid volume is approx. half the listed value)</li><li>Maximum Inlet-to-Outlet Pressure: 690 kPa</li><li>Wetted Materials: 316 stainless steel, PTFE, zirconium silicate, Kalrez, Inconel, and 26-1 stainless steel – all selected for chemical resistance and long-term durability</li></ul><h5>Temperature Control</h5><ul><li>Operating Range: 4 °C to 70 °C</li><li>Stability: Maintains temperature within &lt; 0.005 °C for precise measurements</li><li>Ambient Conditions: 0 °C to 40 °C; relative humidity 20% to 70%</li></ul><h5>Electronics and Connectivity</h5><ul><li>Digital Output: 20-bit resolution at 10 Hz for high-fidelity data</li><li>Auxiliary I/O:<ul><li>Inputs: Autoinject contact closure, Alarm In, and 2 analog inputs (18-bit, ±10 V)</li><li>Outputs: Autoinject retransmit, Alarm Out, Recycle Out</li></ul></li><li>Communication Interface: Ethernet for seamless integration into lab networks and data systems</li></ul><h5>User Interface</h5><ul><li>Front Panel Display: 7” (178 mm) high-resolution capacitive touchscreen for monitoring signals, diagnostics, and adjusting settings with ease</li></ul><h5>Instrument Dimensions</h5><ul><li>Size: 58 cm (L) × 36 cm (W) × 18 cm (H)</li></ul><h4>Other Viscometers</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1267">microViscoStar™ </a>- Similar to the ViscoStar but designed specifically for use with UHPLC/APC.</li></ul>

Waters/Wyatt microViscoStar viscometer

The Waters microViscoStar incorporates multiple novel technologies to provide the highest sensitivity, stability and solvent compatibility of any available viscometer for UHPLC. 

<h4>Highly Sensitive</h4>A highly sensitive, on-line differential viscometer used with SEC-MALS-IV to determine the size and conformation of polymers, proteins and peptides separated by UHPLC.<h4>Multiple Novel Technologies</h4>The microViscoStar™ incorporates multiple novel technologies to provide the highest sensitivity, stability and solvent compatibility of any available viscometer for UHPLC. Its ease-of-use and serviceability make it the perfect companion for Wyatt's microDAWN™ light scattering and microOptilab™ refractive index detectors for UHPLC.<h4>Another path to polymer and protein characterization: SEC-MALS-IV</h4>When coupled with size-exclusion chromatography and a concentration detector such as the Optilab™ , the microViscoStar derives the intrinsic viscosity (IV) of polymers. An inherently valuable physico-chemical property in itself, IV is best utilized together with molar mass information from multi-angle light scattering (SEC-MALS-IV) to derive the hydrodynamic radius, conformation, and branching ratio of a macromolecule in solution.<img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Waters_Visco_Star_pump_pulse_suppression_with_inset_e8c2fbfc18.jpg" alt="Waters microViscoStar: Patented pulse compensation technology in the microViscoStar eliminates pump fluctuations. At the hardware level, the bridge is impedance-matched for dynamic balance, greatly reducing the effect of pump pulses. An additional layer of software filtering eliminates residual pump pulses." srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Waters_Visco_Star_pump_pulse_suppression_with_inset_e8c2fbfc18.jpg 233w," sizes="100vw" width="348" height="233"><h5>Key benefits of the ViscoStar</h5><ul><li>High sensitivity, equivalent to 5 ng of 100 kDa polystyrene in THF at 1.0 mL/min injected onto a standard GPC column</li><li>High dynamic range of over 135,000:1. The full measurement range corresponds to over 13 mg/mL of 100 kDa polystyrene in THF</li><li>Extended solvent compatibility with all-316 stainless steel transducers</li><li>Temperature range from 4 °C to 70 °C for polymers or proteins that require a range of temperatures</li><li>Minimal band broadening thanks to fast pressure transducers</li><li>Pump pulse suppression to below 4 Pa (hardware alone) or 1 Pa (hardware + software) without loss of chromatographic resolution</li><li>Automated capillary bridge tuning with no moving parts, to guarantee a perfectly balanced bridge, every run</li><li>Enhanced thermal stability with drift of &lt;1.25 Pa/hour</li></ul><h4>SEC-IV without MALS?</h4>Yes, as much as we love MALS for optimal polymer analysis, it's true that there are occasions when it just won't fit the bill.Universal calibration is a parallel technique to SEC-MALS for determining molecular weight, commonly used to characterize linear polymers. While universal calibration overcomes some of the problematic issues associated with column calibration by reference molecules, it does not account for non-ideal column behavior caused by chemical solute-column interactions or hyperbranching.Universal calibration is particularly-well suited to replace MALS for:<ul><li>Strongly fluorescent polymers</li><li>Weakly-scattering polymer/solvent systems with very low dn/dc values</li></ul>The Mark-Houwink-Sakurada relationship makes use of empirical relationships between intrinsic viscosity and molar mass, specific to each polymer and solvent, to estimate molar mass from SEC-IV measurements. Of course, the best way to determine MHS coefficients is to measure them using SEC-MALS-IV and ASTRA™<h4>Specifications</h4><h5>Measurement Capabilities – Differential Viscosity</h5><ul><li>Sensitivity: Detects as little as 5 ng of 100 kDa polystyrene in THF at 1 mL/min under typical UHPLC-SEC conditions.</li><li>Differential Pressure Performance:<ul><li>Noise: &lt; 0.05 Pa</li><li>Range: Up to 6.9 kPa, with overpressure protection</li><li>Drift: &lt; 1.25 Pa/hr</li><li>Linearity: Within 0.5% of full scale</li><li>Dynamic Range: 135,000:1</li></ul></li><li>Capillary Bridge Autobalance: Thermal, leak-free mechanism ensures stable baseline performance.</li><li>Sample Shear Rate: Up to 15,000 Hz with water at 1 mL/min for high-resolution viscosity profiling.</li></ul><h5>Fluidics and Compatibility</h5><ul><li>Maximum Flow Rate: Aqueous and THF: up to 1 mL/min (Note: Using an inline UV detector may reduce allowable flow rate.)</li><li>Delay Volume: 5.4 mL</li><li>Maximum Inlet-to-Outlet Pressure: 3000 kPa (435 psi)</li><li>Wetted Materials: 316 stainless steel, PTFE, zirconium silicate, Kalrez®, Inconel, 26-1 stainless steel, and nickel</li></ul><blockquote>Note: Due to the location of wetted components, this instrument is primarily recommended for use with organic solvents. High-ionic-strength buffers may cause corrosion.</blockquote><h5>Temperature Control</h5><ul><li>Operating Range: 4 °C to 70 °C</li><li>Stability: Maintains temperature within &lt; 0.005 °C</li><li>Ambient Conditions: 0 °C to 40 °C; 20% to 70% relative humidity</li></ul><h5>Electronics and Connectivity</h5><ul><li>Digital Output: 20-bit resolution at 10 Hz</li><li>Auxiliary I/O:<ul><li>Inputs: Autoinject contact closure, Alarm In, and 2 analog inputs (16-bit, ±10 V)</li><li>Outputs: Autoinject retransmit, Alarm Out, Recycle Out</li></ul></li><li>Communication Interface: Ethernet</li></ul><h5>User Interface</h5><ul><li>Front Panel Display: 7” (178 mm) high-resolution, capacitive multi-touch screen for real-time monitoring, configuration, and diagnostics</li></ul><h5>Instrument Dimensions</h5><ul><li>Size: 60 cm (L) × 36 cm (W) × 17 cm (H)</li></ul><h4>Other Viscometers</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1117">ViscoStar™</a> - A highly sensitive, on-line differential viscometer used in conjunction with SEC-MALS to determine the size and conformation of all types of biopolymers, synthetic polymers and even proteins and peptides.</li><li>The ViscoStar incorporates multiple novel technologies to provide the highest sensitivity, stability and solvent compatibility of any available viscometer for GPC. Its ease-of-use and serviceability make it the perfect companion for Wyatt's DAWN® light scattering and Optilab® refractive index detectors. Temperature-controlled from 4 °C to 70 °C .</li></ul>

Waters/Wyatt Optilab On-line Differential Refractometer

The Waters Optilab is a unique dRI detector for standard HPLC/GPC with 256 times the detection power and 50 times the dynamic range of every other RI detector for chromatography in existence today, making it a valuable addition to any MALS setup.

<h4>Independent of Chromaphores</h4>Independent of chromaphores or fluorophores, differential Refractive Index (dRI) detectors serve as accurate and versatile concentration detectors in all types of solvents. dRI is part of standard instrumentation for a variety of macromolecular characterization techniques including standard chromatography, SEC-MALS, FFF-MALS and CG-MALS.<h4>Unique dRI Detector</h4>The Optilab is a unique dRI detector for standard HPLC/GPC with 256 times the detection power and 50 times the dynamic range of every other RI detector for chromatography in existence today, making it a valuable addition to any MALS setup.<h4>Product Description</h4>Using a combination of cutting-edge semiconductor photodiode technology and proprietary computer algorithms, the Optilab achieves an unprecedented combination of sensitivity and range. These features mean that it addresses both standard chromatographic applications and some challenges unique to light scattering measurements such as high concentrations, determination of sample refractive increments (dn/dc) and solvent refractive index.<h5>Key Benefits</h5><ul><li>Extraordinary dRI measurement range, -0.0047 to +0.0047 RIU (comparable to 25 mg/mL protein in PBS)</li><li>Highly sensitive, ±7.5 x 10 -10 &nbsp;RIU across the entire measurement range</li><li>Measures solvent absolute refractive index, required for MALS calculations</li><li>Measures at the same wavelength as the MALS detector, for maximum accuracy in MALS calculations</li><li>Band broadening &lt; 20 µL preserves SEC resolution</li><li>Temperature range 4 °C to 65 °C for proteins and polymers that require a range of temperatures</li><li>High-resolution, touch-screen front panel displays data graphs and diagnostics, even with the host computer turned off</li></ul><h5>Unparalleled Features</h5>Even though the Optilab offers a dRI measurement range orders of magnitude beyond that of other dRI detectors, unlike other detectors there are no range or gain settings in the Optilab. The full range of instrument detection is always present, and the full sensitivity exists over the entire range.The Optilab can also do something no other on-line refractive index detector can: it can measure the absolute refractive index of a solution. In addition, it can measure the dn/dc of a solvent at the same wavelength of light as the light scattering instrument. And just to make sure it can't be matched, you can replace the light source in the Optilab for other wavelengths (e.g., 690, 633, 488, etc.) in just minutes!The Optilab can go below ambient temperatures as easily as above ambient. No hassle temperature control can be programmed from 4 °C to 65 °C. Compatible with all HPLC systems.<h4>Specifications</h4><h5>Measurement Capabilities – Differential Refractive Index</h5>Standard Optilab Configuration<ul><li>Measurement Range: –0.0047 to +0.0047 RIU</li><li>Peak-to-Peak Noise: ±7.5 × 10⁻¹⁰ RIU (Measured per ASTM-E1303-95 at 25 °C)</li></ul>High Concentration (HC) Option<ul><li>Measurement Range: –0.0026 to +0.034 RIU</li><li>Peak-to-Peak Noise: ±1.5 × 10⁻⁹ RIU (Optimized for higher concentration samples)</li><li>Response Time: User-selectable from 0.1 to 5 seconds</li><li>Absolute Refractive Index Measurement Range: 1.2 to 1.8 RIU</li><li>Absolute RI Sensitivity: ±0.002 RIU</li></ul>Fluidics and Flow Properties<ul><li>Band Broadening: &lt;12 µL</li><li>Fill Volume (Inlet Tubing + Flow Cell): 51.4 µL</li><li>Outlet Tubing Volume (0.030" ID): 370 µL</li></ul>Pressure Ratings<ul><li>Inlet Back Pressure (0.5 mL/min H₂O @ 20 °C): &lt;22 psi (1.5 bar)</li><li>Max Outlet Back Pressure: 30 psi (2 bar)</li></ul>Temperature Control<ul><li>Sample Temperature Control Range: 4 °C to 65 °C</li><li>Regulation Stability: ±0.005 °C</li></ul>Light Source Options<ul><li>Standard: 660 nm</li><li>HC Model (Optional): 488 nm, 785 nm, or 940 nm</li></ul>Safety Features<ul><li>Built-in vapor and liquid leak sensors for enhanced lab safety</li></ul>Electronics and Connectivity<ul><li>Digital Output: 22+ bit resolution @ 10 Hz</li><li>Analog Output: ±10 V or ±1 V, 18-bit resolution</li><li>Computer Interface: Ethernet</li></ul>Auxiliary I/O Ports<ul><li>Inputs:<ul><li>Analog In (16-bit, ±10 V A/D)</li><li>Zero dRI</li><li>Purge</li><li>Recycle In</li><li>Autoinject In</li><li>Alarm In</li></ul></li><li>Outputs:<ul><li>Autoinject Retransmit</li><li>Alarm Out</li><li>Recycle Out (controls Orbit™ valve switching)</li></ul></li></ul>User Interface<ul><li>Front Panel Display: 7” (178 mm) high-resolution multi-touch screen for viewing signals, diagnostics, and adjusting instrument settings</li></ul>Instrument Dimensions<ul><li>Size: 58 cm (L) × 36 cm (W) × 18 cm (H)</li></ul><h4>Other Refractive Index Detectors</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1269">microOptilab™</a> - The first RI detector specifically designed for use with all UHPLC systems.</li></ul>

Waters/Wyatt microOptilab On-line Differential Refractometer

The Waters microOptilab brings the universal detection capabilities, unsurpassed concentration range, and superb sensitivity and linearity

<h4>Independent of Chromaphores</h4>Independent of chromaphores or fluorophores, differential Refractive Index (dRI) detectors measure concentrations of all types of samples in almost all solvents. The microOptilab brings the universal detection capabilities, unsurpassed concentration range, and superb sensitivity and linearity – familiar from the Optilab™ – to the domain of UHPLC.<h4>SEC-MALS with UHPLC</h4>Like standard SEC, UHPLC-SEC analysis relying on column calibration with molecular standards is unreliable when the standards do not accurately reflect your samples in terms of conformation, non-ideal (e.g., hydrophobic) column interactions or reversible association. SEC-MALS employs a microDAWN™ MALS detector downstream of the SEC column and UV detector, to determine molar mass / molecular weight distributions of macromolecules independently of column calibration.Refractive index detection is an important component of any SEC-MALS system, and necessary for analyzing conjugated proteins or polymers devoid of chromaphores. The microOptilab connects with the microDAWN MALS detector for a complete UHP-SEC-MALS setup that transfers the many benefits of SEC-MALS analysis to the realm of UHPLC.<h5>Key benefits</h5><ul><li>Very low band broadening, &lt; 4 µL, to maintain the resolution of very narrow UHPLC peaks</li><li>Extraordinary dRI measurement range, -4.7 x 10 -3 &nbsp;to +4.7 x 10 -3 &nbsp;RIU (comparable to 25 mg/mL protein in PBS)</li><li>Highly sensitive, ± 1.5 x 10 -9 &nbsp;RIU across the entire measurement range</li><li>Measures solvent refractive index, require for MALS calculations</li><li>Measures at the same wavelength as the MALS detector, for maximum accuracy in MALS calculations</li><li>Temperature range 4 °C to 65 °C for proteins and polymers that require a range of temperatures</li><li>High-resolution, touch-screen front panel displays data graphs and instrument diagnostics, even with the host computer turned off</li></ul><h5>Unparalleled features</h5><ul><li>Considering the extremely small peak volumes of UHPLC, band broadening in the microOptilab is kept to the absolute minimum, at less than 4 µL, as compared to 20 - 40 µL or more for standard RI detectors. As a result, the superior resolution from UHPLC columns and instruments will not be compromised with the microOptilab RI detector.</li><li>Even though the microOptilab offers a dRI measurement range orders of magnitude beyond that of other UHPLC dRI detectors, unlike other detectors there are no range or gain settings in the microOptilab. The full range of instrument detection is always present, and the full sensitivity exists over the entire range. The small flow cell and proprietary temperature regulation enable stable RI baselines and signals which further enhance the detector's sensitivity.</li><li>Unlike all other dRI detectors, the microOptilab also measures absolute refractive index in order to determine solvent RI, necessary in light scattering calculations and useful in assessing the accuracy of mixing of solvent gradients.</li><li>The microOptilab can be operated below ambient temperatures as easily as above ambient. No hassle temperature control can be programmed down to 4 °C or as high as 65 °C.</li><li>The microOptilab is compatible with all UHPLC systems.</li></ul><h4>Specifications</h4><h5>Measurement Capabilities – Differential Refractive Index</h5><ul><li>Differential RI Measurement Range: –0.0047 to +0.0047 RIU</li><li>Peak-to-Peak Noise: ±1.5 × 10⁻⁹ RIU (Measured per ASTM-E1303-95 at 25 °C with a 0.5 sec collection interval)</li><li>Response Time: User-selectable from 0.04 to 5 seconds</li><li>Absolute Refractive Index Range: 1.2 to 1.8 RIU</li><li>Absolute RI Sensitivity: ±0.002 RIU</li></ul>Fluidics and Flow Properties<ul><li>Band Broadening: &lt;4 µL (at 0.3 mL/min)</li><li>Fill Volume (Inlet Tubing + Flow Cell): 13.7 µL</li><li>Outlet Tubing Volume (0.030” ID): 228 µL</li></ul>Pressure Ratings<ul><li>Inlet Back Pressure (0.5 mL/min H₂O @ 20 °C): &lt;320 psi (22 bar)</li><li>Maximum Permissible Outlet Pressure: 30 psi (2 bar)</li></ul>Temperature Control<ul><li>Control Range: 4 °C to 65 °C</li><li>Stability: ±0.005 °C for precise thermal regulation</li></ul>Optical Source and Safety<ul><li>Light Source: 660 nm wavelength</li><li>Safety Sensors: Integrated vapor and liquid leak detection</li></ul>Electronics and Connectivity<ul><li>Digital Output Rate: 25 Hz</li><li>Analog Output: ±10 V or ±1 V, 18-bit resolution</li><li>Computer Interface: Ethernet for seamless data connectivity</li></ul>Auxiliary I/O Ports<ul><li>Inputs:<ul><li>Analog In (16-bit, ±10 V A/D)</li><li>Zero dRI</li><li>Purge</li><li>Recycle In</li><li>Autoinject In</li><li>Alarm In</li></ul></li><li>Outputs:<ul><li>Autoinject Retransmit</li><li>Alarm Out</li><li>Recycle Out (Orbit™ valve switching)</li></ul></li></ul>User Interface<ul><li>Display: 7” (178 mm) high-resolution multi-touch screen for viewing live signals, diagnostics, and configuring instrument settings</li></ul>Instrument Dimensions<ul><li>Size: 58 cm (L) × 36 cm (W) × 18 cm (H)</li></ul><h4>Other Refractive Index Detectors</h4><ul><li><a target="_blank" rel="noopener noreferrer" href="https://lcms.labrulez.com/products/1268">Optilab™</a> - A unique on-line differential refractometer for measuring concentration of any macromolecule, regardless of chromophores. Temperature controlled from 4 °C to 65 °C. The high-concentration option accommodates protein concentration up to 180 mg/mL.</li></ul>

Watrex SCI-AQUA ROWAPUR BASIC Water Purification System

Fully automatic system for the preparation of pure water based on reverse osmosis. Reverse osmosis membrane is protected by three-stage pre-filtration consisting of sediment filter and two carbon adsorption filters.

Fully automatic system for the preparation of pure water based on reverse osmosis. Reverse osmosis membrane is protected by three-stage pre-filtration consisting of sediment filter and two carbon adsorption filters.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/sci-aqua-basic/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex SCI-AQUA BASIC</a> is driven by water pressure from water distribution network, it does not require electric power. It prepares water free of mechanical impurities, chlorine, 95-99% of all dissolved substances are removed.&nbsp;Depending on input water composition and supply pressure the output water has a specific conductivity in the range of 10-20 µS/cm.Price: <bdi>384&nbsp;€</bdi> ex. VAT<h4>Features</h4><ul><li>Removes 95 – 99% of dissolved substances</li><li>Typical specific conductivity of produced water 10 – 20 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Daily production capacity: 50 – 70 liters</li><li>Input water pressure: 0,3 – 0,7 MPa</li><li>Input water temperature: &lt; 35 °C</li><li>Maximum input water TDS: 1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

Watrex SCI-AQUA ROWAPUR DEMI 90 Water Purification System

Fully automatic system for the preparation of clean water based on reverse osmosis combined with purification by a mixed ion exchanger.

Fully automatic system for the preparation of clean water based on reverse osmosis combined with purification by a mixed ion exchanger. Reverse osmosis membrane is protected by two-stage pre-filtration consisting of sediment and carbon filter.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/rowapur-demi-90/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex ROWAPUR DEMI</a> is driven by water pressure from water distribution network, it does not require electric power.The output water has a specific conductivity less than &lt;1 µS/cm. The system includes a conductivity tester with optical indication of output water quality.Price: <bdi>761 €</bdi> ex. VAT<h4>Features</h4><ul><li>Specific conductivity of produced water &lt;1 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>1000 – 2000 liters polishing filter capacity</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Specific conductivity of produced water: &lt; 1 µS/cm</li><li>Daily production capacity:&nbsp;up to 90 liters</li><li>Input water pressure:&nbsp;0,3 – 0,7 MPa</li><li>Input water temperature:&nbsp;&lt; 35 °C</li><li>Maximum input water TDS:&nbsp;1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

Watrex SCI-AQUA ROWAPUR DEMI 180 Water Purification System

Fully automatic system for the preparation of clean water based on reverse osmosis combined with purification by a mixed ion exchanger. Reverse osmosis membrane is protected by two-stage pre-filtration consisting of sediment and carbon filter.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/rowapur-demi-180/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex ROWAPUR DEMI</a> is driven by water pressure from water distribution network, it does not require electric power.The output water has a specific conductivity less than &lt;1 µS/cm. The system includes a conductivity tester with optical indication of output water quality.Price: 833<bdi> €</bdi> ex. VAT<h4>Features</h4><ul><li>Specific conductivity of produced water &lt;1 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>1000 – 2000 liters polishing filter capacity</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Specific conductivity of produced water: &lt; 1 µS/cm</li><li>Daily production capacity:&nbsp;up to 180 liters</li><li>Input water pressure:&nbsp;0,3 – 0,7 MPa</li><li>Input water temperature:&nbsp;&lt; 35 °C</li><li>Maximum input water TDS:&nbsp;1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

Watrex SCI-AQUA ROWAPUR 090 Water Purification System

Fully automatic system for the preparation of pure water based on reverse osmosis.

Fully automatic system for the preparation of pure water based on reverse osmosis. Reverse osmosis membrane is protected by three-stage pre-filtration consisting of sediment filter and two carbon adsorption filters.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/rowapur-90/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex ROWAPUR</a> is driven by water pressure from water distribution network, it does not require electric power.It prepares water free of mechanical impurities, chlorine, 95-99% of all dissolved substances are removed. Depending on input water composition and supply pressure the output water has a specific conductivity in the range of 10-20 µS/cm.Price: 500<bdi>&nbsp;€</bdi> ex. VAT<h4>Features</h4><ul><li>Removes 95 – 99% of dissolved substances</li><li>Typical specific conductivity of produced water 10 – 20 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Daily production capacity:&nbsp;up to 90 liters</li><li>Input water pressure:&nbsp;0,3 – 0,7 MPa</li><li>Input water temperature:&nbsp;&lt; 35 °C</li><li>Maximum input water TDS: 1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

Watrex SCI-AQUA ROWAPUR 180 Water Purification System

Fully automatic system for the preparation of pure water based on reverse osmosis. Reverse osmosis membrane is protected by three-stage pre-filtration consisting of sediment filter and two carbon adsorption filters.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/rowapur-180-reverse-osmosis-system/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex ROWAPUR</a> is driven by water pressure from water distribution network, it does not require electric power.It prepares water free of mechanical impurities, chlorine, 95-99% of all dissolved substances are removed. Depending on input water composition and supply pressure the output water has a specific conductivity in the range of 10-20 µS/cm.Price: 541<bdi>&nbsp;€</bdi> ex. VAT<h4>Features</h4><ul><li>Removes 95 – 99% of dissolved substances</li><li>Typical specific conductivity of produced water 10 – 20 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Daily production capacity:&nbsp;up to 180 liters</li><li>Input water pressure:&nbsp;0,3 – 0,7 MPa</li><li>Input water temperature:&nbsp;&lt; 35 °C</li><li>Maximum input water TDS: 1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

Watrex SCI-AQUA ROWAPUR 270 Water Purification System

Fully automatic system for the preparation of pure water based on reverse osmosis. Reverse osmosis membrane is protected by three-stage pre-filtration consisting of sediment filter and two carbon adsorption filters.<a target="_blank" rel="noopener noreferrer" href="https://watrex.com/obchod/water-purification/reverse-osmosis-systems/rowapur-270-reverse-osmosis-system/?utm_source=labrulez&amp;utm_medium=propagace&amp;utm_campaign=labrulez_produkty">Watrex ROWAPUR</a> is driven by water pressure from water distribution network, it does not require electric power.It prepares water free of mechanical impurities, chlorine, 95-99% of all dissolved substances are removed. Depending on input water composition and supply pressure the output water has a specific conductivity in the range of 10-20 µS/cm.Price: 592<bdi>&nbsp;€</bdi> ex. VAT<h4>Features</h4><ul><li>Removes 95 – 99% of dissolved substances</li><li>Typical specific conductivity of produced water 10 – 20 µS/cm</li><li>6 to 12 months filters lifetime</li><li>2 – 5 years RO membrane lifetime</li><li>No electricity is required for operation</li><li>Supplied from water distribution network</li></ul><h4>Technical specification</h4><ul><li>Daily production capacity:&nbsp;up to 270 liters</li><li>Input water pressure:&nbsp;0,3 – 0,7 MPa</li><li>Input water temperature:&nbsp;&lt; 35 °C</li><li>Maximum input water TDS: 1200 mg/l</li></ul><img src="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/Watrex_Water_purification_systems_8810a2c48a.jpg" alt="Watrex: Water purification systems" srcset="https://storage.googleapis.com/labrulez-bucket-strapi-h3hsga3/thumbnail_Watrex_Water_purification_systems_8810a2c48a.jpg 245w," sizes="100vw" width="2286" height="1130">Basic requirements for laboratory operations include the easy availability of pure laboratory water in the necessary quantity and quality. Distillation, a popular technique for producing purified water in the past, has virtually completely supplanted by less energy-intensive and operationally dependable alternatives today. Reverse osmosis is the foundation of the majority of contemporary systems for producing purified water; in reality, this principle is supplemented with adsorption, ion exchange, filtration, ultrafiltration, and other physical-chemical techniques.

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