Qualification FAQ – MicroSolv Technology Corporation

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Are column oven temperatures qualified by the HSQ Kit for HPLC?

No, the use of column ovens are not required for PQ Testing of your HPLC instruments.

Can I choose HPLC mobile phase percentages other than those used in the HSQ Spreadsheet?

The HSQ Kit gradient test calls for mobile phase compositions of 100/0% and 90/10% (for each of up to four solvent pairs). The user then inputs the absorbance values at each portion of the gradient into the HSQ Spreadsheet, and the software then automatically calculates the pump accuracy. In theory, percentages other than 90/10 could be used, […]

Can I measure the extra column volume of my HPLC system and compare it to extra column volume of another instrument?

Yes, that is one of the added features of the MicroSolv HSQ Kit™. Using the Rs Test Mixture, and the Excel template, using sophisticated algorithms, a very good estimation of your instruments extra column volume is easily determined. To compare it to another instrument, you must use the same column under the exact same conditions […]

Can I use qualification solutions with different lot numbers?

If you require or would like to use a mix of different lot numbers for solutions from the MicroSolv HSQ™, UV-vis™ or PQ™ Kit solutions, this is an acceptable protocol under some conditions: You cannot mix different lots among the L1-L6 solutions. These solutions are for UV detector linearity and are designed to be used as a set. However, you can for […]

Can I use the holmium oxide solution (PQA-HOX-01) in a UV instrument that calls for Starna or NIST cuvettes?

If your instrument manual calls for the SRM 2034 Holmium Oxide (HOX) Solution, you can use our HOX solution instead. The SRM 2034 item has been discontinued by NIST but our HOX solution is fully NIST traceable to their published standard. Therefore our HoX solution will be entirely suitable for this purpose. Holmium Oxide Solution Product […]

Can I use the HSQ Kit for IQ and OQ as well as PQ?

From USP <1058>: Installation Qualification (IQ) – The documented collection of activities necessary to establish that an instrument is delivered and designed as specified, and is properly installed in the selected environment, and that this environment is suitable for the instrument. Operational Qualification (OQ): The documented collection of activities necessary to demonstrate that an instrument will function according […]

Can I use the HSQ Kit with the Longer Column from the Original PQ Kit? – FAQ

If you’re required to use the older 150mm PQ Kit column and cannot use the 75mm HSQ Kit Column, you can adjust the parameters in the Excel Spreadsheet shown here: Change Flow Rate = 1 mL/minute Change Expected Retention Times/Run Times = 4x longer than in the Excel Sheet to compensate for lower flow rate (see […]

Can I use the MicroSolv HSQ Kits for HPLC to qualify a UHPLC Instrument?

Fundamentally, UHPLC is no different from conventional HPLC. All of the same principles apply, only UHPLC uses higher pressure pumps, lower extra-column system volume, and so on. You can even use the same standard 4um column to qualify the UHPLC instrument. Whether you are qualifying a UHPLC or HPLC instrument, the Kit provides the same […]

Can I Use the Standard, HPLC Column Supplied in the HSQ Kit for UHPLC Instrument Qualification? – FAQ

Yes, the MicroSolv HSQ Kit is supplied with a 5um Particle Size HPLC Column that can be used to qualify a UHPLC instrument; you do not need a 2um particle size in a UHPLC column. Many qualification parameters are detector related such as Absorbance Linearity, Wavelength Accuracy, etc. Column-related aspects are not involved in the […]

Can I use the UV Calibration Kit to Qualify Detection at 200nm? – FAQ

The Caffeine Standard provides two data points at 205nm and 273nm. The Holmium Oxide Solution covers 14 absorbance bands in the range of 241nm – 641nm. Hence, you can obtain a calibration curve from these various data points and then extrapolate down to 200nm. This is the lowest you would need to go because of […]

Can the HSQ Kit for HPLC be used to measure the LOQ and LOD differences between instruments in my lab?

Yes, this is another really great “extra feature” of the HSQ Kit™. By determining the, signal to noise ration, the extra column volumes and the dwell volume of each instrument using the HSQ Kit and the software provided, you can determine the differences in LOQ (limits of quantitation) and LOD (limits of detection) between instruments. […]

Can the HSQ or PQ kits be used with Waters equipment and Empower chromatography data software?

Yes, the MicroSolv HSQ™ and PQ™ kits can be used with any type of HPLC instrument or data system the instrument may have. However, note that Waters Associates™ offers a somewhat similar performance qualification kit to the MicroSolv kits that is designed to run automatically with and under Empower™, automating the entire process. The MicroSolv PQ kit […]

Can the L3 Linearity Solution Supplied in the HSQ Kit be used for UV Wavelength Accuracy? – FAQ

The short answer is yes. Many other Qualification vendors use only Caffeine for Wavelength Accuracy, at 205nm and 273nm. Holmium oxide of course covers 241-641nm, and is directly traceable to NIST standards. However, if you want, there is no reason you could not simply take the spectrum of Caffeine as well. The L3 Solution provides […]

Do the different tests in the HSQ Kit for HPLC use the same method conditions?

Yes, this is why it can be run overnight from an autosampler. Simply set your conditions, program the system for PQ Validation and let it run. Use the following Conditions: Wavelength: 273nm Injection Volume: 10µl (except for volume linearity) Flow Rate: 1 mL/minute Temperature: Ambient

Do you supply an SOP template with the HSQ Qualification Kit

We no longer supply a formal template or extensive written SOP for Performance Qualification. However, the Excel template that is provided with the HSQ (High Speed Qualification) kit can be a model for your SOP taking into account the tests that you will do and the objective of each test. Also, the Quick Start Instructions […]

Does the HSQ kit for HPLC work with a Jasco HPLC? Can all tests be performed with a 20 µL micro-loop?

It depends. With the Agilent 1100 type systems for example, a 10 µL injection volume will produce a maximum peak height for the L6 solution of typically about 1600 – 1800 mAU, which is about the maximum absorbance one would use on that detector. For your Jasco system, you would have to try to inject the […]

Does the HSQ Kit Qualify Quaternary HPLC Pumps? – FAQ

Yes, the HSQ Kit™ will work on many types of pumps; from single, binary, and also quaternary pumps. For Gradient Qualification Tests, the base method is called “GRDAB” for “Gradient, pump A/B*”, where the “*” designates that the B fluid circuit is “visualized” with the addition of the Gradient Visualization Solution (GVS). For ternary or […]

Does the method used in the HSQ Kit for HPLC need to be validated?

Validation of the method would not be applicable since it is used to qualify the instrument itself. Suppose you’re talking about LOD or LOQ of the HPLC method as a validation parameter, for example. You wouldn’t need to validate this parameter in the HSQ Kit method because its calculation first assumes that the detector is […]

Does the UV-Vis Calibration Kit Operate in all Three Ranges, UV, Vis and Vis/NIR? – FAQ

Not all Three… The MicroSolv UV Calibration Kit contains Potassium Dichromate for Absorbance Accuracy, Holmium Oxide for Wavelength Accuracy and Potassium Iodide for Stray Light measurements. It will not cover the other Wavelength Ranges and can be used to comply with USP <857> for the ranges where the supplied solutions are applicable. The Chart below […]

Fluctuations in readings when using the UV-Vis Calibration Kit

Fluctuations in readings in a UV-vis instrument are almost certainly due to a failing lamp. A typical insturment which uses a xenon lamp is normally rated for only about 500 hrs, while a deuterium can range from 1,000-3,000 hrs. However, they will sooner or later. If you are not sure of the hours on the lamp, I would […]

How can I make changes to locked cells in the Excel spreadsheet that comes with the HSQ Kit?

The spreadsheet is locked against changes so that only end-user data cells may have values entered. In order to alter locked cells, the password will be required. If you need to make such changes, the password can be provided. However, at this point, the spreadsheet would no longer be considered validated. If you are not strict GMP, please inquire […]

How can I obtain qualification solutions with a longer expiration date than normal?

All of the qualification solutions start with a 2 year expiration date from the time they were made and we have validated data to support this expiration. The date of manufacture and the expiration date are both printed on the COA provided with the product. You can use these solutions with full confidence right up to […]

How can I use the HSQ Kit to perform qualification for my specific analytes?

The MicroSolv Qualification Kits are designed to qualify instruments and not analytes. What you may want be thinking of is system suitability which is different from Performance Qualification (PQ) and our kits are not for that use. Once you qualify the instrument, it doesn’t matter what types of analytes you are working with since the […]

How do I clean the PQ or HSQ Kit™ Column?

“Since the column is used only for test purposes, it should never become contaminated, and should require only a minimal 10-15 minute equilibration prior to starting the sequence. If it ever requires cleaning, flush for 15-30 minutes with 50:50 acetonitrile : water.” —User manual, Rev. 1.08, section II.3, pg. 9. PQ & HSQ Kits for […]

How do I determine wavelength accuracy below 230nm for HPLC instruments?

Holmium Oxide is an internationally recognized wavelength calibration primary standard, supported by NIST and other standards organizations. It provides 14 official absorbance bands ranging from 241 nm to 641 nm. Caffeine is considered a secondary standard, with two absorbance bands at 205 nm and 273 nm. Caffeine is also widely used as a wavelength and […]

How do I measure pump flow accuracy in HPLC?

Use a dry 5ml volumetric flask and a calibrated stopwatch to confirm the flow accuracy at 1.0 mL/minute. Note that this flow rate check is not meant to replace the typically more extensive Operational Qualification checks at several flow rates. Rather it is to document the suitability of the pump to perform various PQ Test protocols described […]

How do I store the HPLC column supplied in the HSQ kit?

One of the best features of the MicroSolv HSQ Kit™ is that the kit was designed for ease and speed. The HPLC Column is shipped and stored in the same mobile phase that is used during the testing. The column is always ready to go with minimal equilibration time. The mobile phase/storage solvents were chosen […]

How do I store the HSQ™ & PQ™ test solutions when not in use?

The test solutions should be stored at room temperature (20-25°C) in a safe, dry place. Never refrigerate these solutions. PQ Kit Home Page

How does a Pulse from an HPLC Pump Affect the Column and its Performance? – FAQ

If your HPLC pump produces a “pulse” (i.e. cyclic fluctuation in flow/pressure), this can cause problems for your Method and Column. The pump will not be delivering optimally Precise/Accurate Flow and this can cause your system to not meet System Suitability Criteria as well as other issues. Sudden changes in the Pressure can also cause […]

How does the shorter column supplied in the HSQ Kit affect the data compared to the longer column from the PQ Kit?

The HSQ Kit comes with a 4.6 x 75mm column that is used in the qualification procedure. The older version of the the HSQ Kit (called the PQ Kit) comes with a 4.6 x 150mm column instead. This difference will not affect the qualification data results and the columns can be used interchangeably. The only difference […]

How many injections should I expect on the HSQ Kit’s HPLC Column?

Thousands… if you do not drop it, overheat it, put too much pressure through it (over 4,000 psi), subject it to pressure shock or inject anything on the column that is not part of the HSQ Kit™.

How many tests will I get out of one HSQ Kit for HPLC?

Depending on how careful you are with transferring the solutions, each kit will produce up to 30 tests. These tests can be on one instrument or on 30 different instruments.

I am getting a different retention time for the peaks in the Resolution Test Mixture on different systems. What impact does this have on the integrity of my data?

No impact at all. There will certainly be mobile phase preparation differences, aging of columns, etc. The HSQ kit instructions call for adjustment of the mobile phase to meet system suitability, which for example in our C of A, calls to adjust the mobile phase to achieve a 4-5 minute retention time window. Keep in […]

I currently use a glass filter, why should I use the HSQ Kit solutions instead when doing qualification of HPLC instruments?

Glass filters for uv detectors are not considered primary standards whereas the solutions in the MicroSolv HSQ Kit™ are primary standards.

Is the Holmium Oxide solution intended as a stand-alone product or solely as a replacement solution for the HSQ Kit?

The Holmium Oxide solution (catalog number PQA-HOX-01 or PQA-HOX-A) can be used as a stand-alone item without having to order the entire HSQ Kit. It is a component of the Kit but can be used to perform wavelength accuracy qualification on its own. You can also order it as a replacement solution if you already have the HSQ […]

Is the HSQ™ Kit complliant with the latest USP-NF chapter revisions for <857> and <1857>?

The MicroSolv HSQ™ Kit materials are NIST traceable. Compliance with USP protocols is demonstrated by the user and not the kits themselves. Specific chapters in the USP refer to procedures and the HSQ™ Kit materials can be used as part of your protocol to meet the relevant USP requirements. For example, the UV and HPLC PQ kits can be used […]

Tip on how to calculate extra-column volume using the MicroSolv HSQ™ Kit software

The extra-column volume test is calculated from the single injection of the Resolution Test Mixture. It uses the RTM method, which is simply the basic PQ™ method, but with a runtime of 3 minutes. The other difference in the RTM method is that is used to acquire the caffeine spectrum, if it is a DAD […]

What are the concentrations used in the Refractive Index Qualification Kit?

The solutions of the HPLC Performance Qualification Solutions for Refractive Index and Light Scattering Detectors have the following certified concentrations: Solution Header 2 L1 0.200 mg/mL ± 0.0002 L2 0.400 mg/mL ± 0.0002 L3 0.500 mg/mL ± 0.0002 L4 1.00 mg/mL ± 0.001 L5 1.50 mg/mL ± 0.001 L6 2.00 mg/mL ± 0.001 […]

What are the differences between the linearity solutions in the HSQ Kit for HPLC?

They are all different, precise concentrations of Caffeine that allow you to easily and accurately determine the linearity of your HPLC UV detector. L3 solution is the only linearity solution that has a Uracil peak present. This peak is used as a pump flow stability marker and has no effect on caffeine or linearity tests.

What are the linearity solutions in the HSQ kit for HPLC made with?

The HSQ Kit’s linearity solutions are made under GMP conditions with calibrated balances and following a strict protocol and comprise Caffeine dissolved in 90:10 Acetonitrile:0.1% Acetic Acid.

What are the peaks/analytes in the Resolution Test Mixture supplied in the HSQ Kit for HPLC?

The peaks/analytes elute in the following order for the PQ Kit™: Uracil Theophylinne Caffeine 8-Chloro-Theophylline For more recent and more efficient HPLC’s, the HSQ Kit™ has a different Resolution Test Mixture with one additional, earlier eluting peak. This additional peak allows the HSQ Software to best determine your system’s Extra Column Volume. This extra peak is not required […]

What is the acceptable wavelength band width for HPLC detectors when performing an HPLC Performance Qualification?

3nm is the normal setting for wavelength bandwidth for HPLC Detectors. 5nm may be considered too wide and 1nm is too narrow for PQ.

What is the difference between the new HSQ Kit™ for HPLC and older PQ Kit™?

The difference between the HSQ Kit™ and PQ Kit™ is mostly speed. HSQ is an acronym for High Speed Qualification. The HSQ Kit™ is supplied with a different, shorter PQ HPLC Column. It is still certified but it is shorter and the run times are much shorter. The time to do the exact same qualification […]

What is the Effect of Extra Column Volume for Different HPLC Instruments on the Same Method? – FAQ

The difference can be very significant. Extra Column Volume (ECV) that is “hidden” in your HPLC system, the fittings, the tubing and the flow cell can greatly impact the efficiency of your method. Extra Column Dispersion (ECD) will result and the smaller the particle, the more impact will be observed. Below are two chromatograms using […]

What is the Gradient Visualization Solution supplied in the HSQ Kit for HPLC?

The HSQ Kit’s Gradient Visualization Solution is made under GMP conditions with calibrated balances and following a strict protocol and comprise 1.0 ml/mL Uracil and Caffeine in 10% Acetonitrile/water with 0.1% Acetic Acid.

What is the mobile phase used in the HSQ Kit for HPLC and how much is used?

For isocratic system testing, prepare one liter of 13:87 Acetonitrile:0.1% Acetic Acid (1mL/L). For gradient testing, prepare 2 liters.