Chromatographic analysis with techniques like HPLC, UHPLC or GC is commonly used in analytical R&D and QC labs to identify, quantify and purify individual components of a mixture. Component purity, concentration and composition have a major influence on final product quality, which can impact product efficiency, safety, shelf-life, and many other important features. To obtain quantitative results, calibration standards or reference substances in known concentrations are needed.
Why is weighing important?
In analytical chromatography, accurate weighing, proper calculation of concentrations and error-free transfer of data have a significant impact on the quality of chromatographic results. Sample preparation can use up to 60% of analytical laboratory time and effort. Moreover, data management such as results transcription and archiving can use up to 25% of working time. Underlying the importance of state-of-the-art process automation and data management: the more tasks that can be automated and simplified without compromising accuracy, the better in terms of conserving resources and enhancing throughput and accuracy.
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Watch the Video: Accurate and Quick Preparation of Standards with the XPR Automatic Balance
Chromatography is a technique commonly used in analytical R&D and QC laboratories. Gravimetric preparation of standard solutions is not only more accurate and reliable than preparation by volumetric methods, but also allows a reduction in the quantities required. With automatic dispensing of powders and liquids, standard preparations for HPLC analysis can be further accelerated. The XPR automatic balance makes standard preparation more efficient and accurate with smaller sample volumes. Watch the video to discover how.
Typical Workflows to Prepare Chromatographic Analysis Samples and Standards
Typical Workflow to Perform an Organic Reaction Optimization
The preparation of samples or reference standards involves similar work steps, whereby the type of procedure depends on the sample in question. Quantitative statements about chromatographic analyses of samples can only be made with the aid of reference substances, so-called calibration standards, or simply standards.
Sample Preparation
Samples must be prepared individually, and dissolved, filtered or chemically extracted if necessary. Sample preparation objectives include:
Bringing analytes into a solution
Separating solid or dissolved interfering substances
Purifying and preconcentrating analytes
Adjusting a concentration for measurement in a suitable solvent
Standard Solution Preparation
The preparation of calibration standards for quantitative analysis is usually easier than sample preparation because the chemicals are available in their purest form, often powdery and soluble. They must be prepared very accurately because they are used to create a calibration curve to calculate sample concentrations. In most cases, multiple concentrations are required, which are often prepared directly before each analysis.
Typical Process Steps
Identify reference standard
Weigh reference standard
Add solvent, shake and mix
Dilute to prepare suitable concentrations
Calculate/record results
Label flasks
Filtrate and transfer to analysis vials
Analysis by a variety of analytical techniques:
HPLC, GC, NMR
ICP-MS, ICP-OES
AA, Organic Elemental Analysis
Titration, Karl Fisher
Data evaluation
Result calculation
Reporting
Secure data storage
Data retrieval (for audit)
Making the Switch to Gravimetric Solution Preparation
Gravimetric sample preparation – weighing both the solid and the solvent – is a state-of-the-art approach to preparing samples and solutions that is specifically described in USP chapter 1251 "Weighing on an analytical balance" in the section "Types of dosing" USP36-NF31 (2013). For more information, consult our White Paper:
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Is chromatographic analysis one of the "time-eaters" in your laboratory? Lay the foundation for your lab of the future and invest in an automated and digitized sample and standard preparation for your chromatographic analyses. Let us show you how the modular elements of our automated solutions – tailored to your specific requirements – can make your chromatography processes more efficient and safer.
Workflow Challenges in Sample and Standard Preparation for Chromatographic Analyses
Manual Volumetric vs. Automated Gravimetric Liquid and Powder Dispensing
Manually weighing out powdery substances with a spatula can be a tedious and challenging task. Especially when amounts are in the low milligram range, it takes skill and experience to weigh accurately.
Even in the best of hands, traditional solution preparation with volumetric glassware is time-consuming and prone to error. An operator will typically weigh a substance onto weighing paper and then transfer it into an Erlenmeyer flask. Does the operator back weigh the paper to ensure no substance is left behind? If not, doubt is created about process accuracy. If so, extra time is added to the workflow. Neither choice is ideal.
With the amount of laboratory time spent processing standard solutions estimated to be greater than 60%, measures that can reduce such incidences can significantly improve overall accuracy and laboratory productivity.
Data Management, Transfer, and Traceability
Data handling and traceability of chromatographic data is laborious and prone to error. For example, a lab technician must be diligent in recording each step of a dataset’s journey when switching among instruments and systems. Especially when a high throughput is necessary and data must be traceable, software-driven workflows and networked systems lead to a modern and highly efficient lab.
Solutions to Speed Up Sample and Standard Preparation for Your Chromatographic Analyses
Accelerate Your Processes — Apply Automation and Digitalization Strategically
Automation is increasingly employed in laboratories to keep productivity high, reduce results subjectivity, and free scientists for high-value experimentation. The tactical adoption of instruments and software aligned with day-to-day procedures can substantially benefit operations.
Where higher throughput is necessary, laboratories can leverage software-driven workflows that integrate instruments into a network and ensure that complete data is collected and securely recorded for every sample.
Here, we provide an overview of our solutions that can help you automate and digitalize your sample and standard preparation for chromatographic analyses. Which one is right for you based on your current chromatographic analyses and throughput requirements?
XPR Analytical Balance — Invest in Your Automated Future
Reliable analytical results begin with accurate weighing. METTLER TOLEDO's XPR Analytical Balances offer readabilities from 0.002 mg to 0.1 mg and a capacities up to 320 g. With a vast range of proven innovative technologies and advanced quality assurance functions, you can be sure of fast, reliable and valid results every time you weigh.
XPR Analytical Balances enable manual and volumetric workflows by means of manual powder solvent dispensing into a volumetric flask. Investing in a XPR Analytical Balance is an excellent start for future automation and digitization of your sample and standard preparation for chromatographic analyses. Many labs consider automation and digitalization prohibitively expensive. With our modular approach, the latest solutions are affordable.
Choose your level of automation and put together your individual digitalized solution. You can either upgrade your analytical balance for accuracy or opt for a more automation and time savings with the XPR Automatic Balance or robotic solution.
The appropriate software can be used at any level of weighing with an XPR balance. Upgrade your sample and standard preparation process and benefit from:
Seamless data management
Fewer steps for greater efficiency
Smaller volumes/sample sizes, less material needed
Higher accuracy and reproducibility
Automation of single or multiple process steps
Automate Data Handling and Documentation
Connect your XPR Balance to LabX™ Balance Software. LabX enables solution preparation with automatic documentation and data processing. Metadata is automatically captured and saved in a centralized database. LabX guides you step-by-step through your SOP with clear on-screen instructions. All calculations are performed automatically and transcription errors are eliminated.
Build a Digital Data Bridge from Your Balance to Your Chromatography System
Add the LabX - OpenLab Plugin to transfer weighing data and metadata from LabX Balance software to the Agilent OpenLab Chromatography Data System (CDS). This Balance–LC/GC bridge lets you greatly reduce error risk, as it eliminates manual calculations and data transcription for the HPLC analysis process.
How to Upgrade Your Workflows in 4 Steps
Step 1: Switch to Accurate Gravimetric Preparation
By making the switch to gravimetric standard preparation, both solids and solvents are measured by weight, not volume. The user defines the concentration required and the target amount of solution. Then, the system software calculates the target amount of solid to dispense and delivers the appropriate amount of diluent according to the actual amount of solid dispense. This assure an extremely accurate and precise concentration.
Add a QLX3 liquid dispensing module to your XPR Analytical Balance, automate the liquid dispensing step and make the switch to the more accurate gravimetric solution preparation with less material required. This is a simple and inexpensive solution for automated calculation and automated gravimetric dispensing of solvents based on manually weighed solid substances (even non-powders).
Gravimetric standard and sample preparation offers the following benefits:
Lower minimum weight
Improved accuracy of prepared solutions and results quality
Fewer workflow steps, better efficiency and productivity
Sample is weighed directly into a vial
Solvent is added accurately based on weight
Less sample, standard, and solvent is needed and less waste is created
Reduced risk of overfilling
No subjective or temperature dependent meniscus reading
Upgrade your balance to an XPR Automatic Balance with powder and liquid dispensing modules and benefit from an automated solution for the gravimetric dispensing of powders and solvents. Automatic weighing reduces the minimum weight by up to 30% and enables material savings of up to 70%, giving you an extraordinarily fast return on your investment.
Step 3: Add a Sample Changer to Automate One-To-Many Powder Dispensing
Add the QS3 sample changer accessory to your XPR Automatic Balance and upgrade it with a semi-automated vial handling to create a solution with increased throughput for up to 30 target vials.
Step 4: Don’t Get Your Hands Dirty — Let the Robot Do the Work
Fully automate your sample and standard preparing processes with the CHRONECT XPR robotic dispensing system and prepare up to 288 solutions in one run, consisting of up to 32 powders and 6 liquids and enabling many-to-many methods. The unique system combines the advanced weighing and dispensing technology of the XPR Automatic Balance with a state-of-the-art 6-axis robotic arm and the easy-to-use CHRONOS software. Reduce time spent at the bench and attend to more valuable tasks.
CHRONECT XPR features fully automated dispensing of powders and liquids, including robotic vial handling. It offers unprecedented accuracy, repeatability, efficiency, and flexibility. Samples and reference standards for chromatographic analyses can be prepared conveniently overnight, making them ready for next steps in the morning.
Application Note: Easy and Cost-Efficient Standard Preparation for HPLC
This application note presents a quick and easy way to prepare accurate reference standards for HPLC and other chromatographic methods. The method enables using less substance and solvent, ultimately significantly saving time and money.
Robotic Preparation of Standards for Chromatographic Analysis
Our application note presents a fully automated, robotic process for preparing accurate samples and reference standards for chromatographic analyses like HPLC. The method allows scientists to reduce time spent at the lab bench and attend to more valuable tasks.
Webinar: Improve Efficiencies in Your Lab with Automated Weighing
Manual weighing processes are time-consuming and labor-intensive, and any mistakes can lead to major errors in downstream processing. Automated weighing addresses these challenges. Our innovative approach to automation saves time and delivers highly reproducible results, giving you the freedom to focus on other critical elements of your analysis.
Chromatography techniques are used to separate mixtures into their constituent parts so that each part can be analyzed separately.
High-performance liquid chromatography (HPLC) can be used to extract individual components from a solution. Components of a mixture separate or elute with HPLC at different times. The eluates from the HPLC column are then fed into various detectors that produce a peak on a graph relative to its concentration as it elutes from the column. The most common type of detector is an ultraviolet-visible spectrometer since most of the substances commonly studied by HPLC, especially pharmaceuticals, exhibit UV absorption.
Gas chromatography (GC) performs the same function as liquid chromatography, but it is used for volatile mixtures. The most universal detector for gas chromatography, with capabilities of both trace quantitative and definitive qualitative information, is the mass spectrometer. The flame ionization detector (FID) is the most popular detector for organic vapors.
What are common terms used in chromatography and what do they meaning?
Glossary of common terms in chromatography:
Chromatography ꟷ Analytical laboratory technique for separating a mixture into its components. The mixture is dissolved in gas or liquid, called the mobile phase, which carries it through a system (a column, a capillary tube, a plate, or a sheet) on which a material, called the stationary phase, is fixed.
Chromatography Detector ꟷ Mass Spectrometry (detector, a mass spectrometer separates ions or molecules according to their mass-to-charge ratio)
MS ꟷ Mass Spectrometry (detector, a mass spectrometer separates ions or molecules according to their mass-to-charge ratio)
FID ꟷ Flame Ionization Detector, frequently used in GC (detection of ions formed during combustion of organic compounds in a hydrogen flame)
CDS ꟷ Chromatography Data System
Sample Preparation ꟷ Preparation of samples for any analysis (often used as a general term for all samples to be analyzed, incl. standards, blanks, etc.)
Standard Preparation ꟷ Preparation of calibration standards for analysis
What are the trends for improving chromatography?
In recent years, much time has been spent on improving analytical speed, resolution, and automation, as well as developing and improving analytical instruments. However, one of the biggest bottlenecks in the process remains sample and standard preparation. It is not uncommon that two-thirds of the analysis time is spent on this process step. Chromatographers are therefore looking for better sample preparation techniques to improve analysis speed and accuracy. They are looking for faster, more cost-effective procedures, and easy-to-use, convenient, and safer methods. Another trend is the increased use of smaller sample sizes. Speeding up or automating sample preparation can reduce analysis time and increase sample throughput while reducing sample sizes. Intelligent interfaces and clever software solutions further accelerate the process and allow error-free data processing and comprehensive report generation.
What is the most frequently used technique for sample preparation?
Weighing is obviously the most popular technique for any sample preparation, as almost every quantitative analysis method requires the measurement of sample weight. But many other preparation techniques are used in analytical laboratories such as dilution, filtration, pipetting, evaporation, concentration, extraction, centrifugation, dying, pH adjustment, grinding, digesting and more.
What is a reference standard?
Reference standards are used to identify or determine the concentration of an analyte in a sample of unknown concentration. The reference standard is prepared with a known concentration, so the unknown sample can be compared to it to determine the actual sample concentration.
How can I make my sample and standard preparation workflow for chromatographic analyses faster and more efficient?
The use of automation systems, like automatic powder and liquid dispensing or liquid handling systems, can greatly improve the throughput of sample and standard preparation processes.
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