academia 12 weighing and analysis in the laboratory news

Weighing and Analysis in the Laboratory

In a country full of large forests and beautiful lakes, the research depart-

ments of the Swedish University of Agricultural Sciences are developing

expertise in biological resources and production. At Umeå, they analyze

the dynamics and management of forest ecosystems by measuring pH,

conductivity, acidity and alkalinity of field samples by titration.

Each week, 50 to 300 samples from vari-ous locations throughout the country are analyzed for any combination of the required parameters in the Department of Forest Ecology and Management. The result quality must remain high, which means that it is crucial for any influences that affect the analyses, i.e. temperature change during a conductivity or pH mea-surement, to be excluded. Furthermore, measurements, such as CO2 degassing, must be in place to avoid pH alteration. Last, but not least, the results have to be available within the shortest time possible to reflect the conditions at the sample’s lake origin.

A titration automation station meets the challengeThe incoming samples are placed onto a

Rondo 30 sample changer. The individual sample is pumped by peristaltic pumps through two heat-jacketed flow-through cells. These cells contain the conductiv-ity and pH sensors. Both parameters are measured separately at a constant tem-perature of 25 +/- 0.5 °C in order to avoid cross-contamination by the electrolyte flow from the pH sensor during the con-ductivity measurement. The sample is then transferred into the titration beaker where the volume is leveled exactly to a pre-defined quantity. Depending on the pH, the T90 Excellence Titrator decides whether alkalinity or acidity shall be ana-lyzed by automatically selecting the ap-propriate titrant. Sample specific method selection, sample identification and re-sult storage are all reliably conducted by LabX® titration software. Field sample

Understanding EcosystemsUsing Multiparameter Analysis

12News

AcademiaAnalytical solutions in the laboratory

Edition No. 1 / February 2013

2 METTLER TOLEDO Academia News 12

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Publisher Mettler-Toledo AG Laboratory Division Im Langacher CH-8606 Greifensee, Switzerland

Production LAB Division MarketingGlobal MarCom Switzerlandwww.mt.com/lab-segmentnews

Subject to technical changes. © Mettler-Toledo AG 02/13 Printed in Switzerland.

containers, closed with self-adhesive alu-minum foils, can be directly placed on the sample changer tray. Sample trans-fer and CO2 degassing, with inert gas, is achieved by penetrating the foil with a needle. Any ambient contamination is completely avoided using this method. Thanks to the thorough cleaning of the system, carry-over from previous samples is non-existent.

SLU Umeå system contributes to environmental research The workstation has been yielding results that perfectly meet the high expectations of Umeå for more than a year now. Thanks to the clever system setup and a complete sample analysis, including cleaning, re-quires a maximum of 10 minutes. The daily sample throughput can be eas-ily analyzed. The important result trend evaluation of these samples is reliably achieved by LabX titration. The system performance can be easily controlled by standards or check samples integrated in-to the workflow. The scientific employees operating the system are very pleased with the simple shortcut operation set-up of the T90 and LabX, especially concerning the simple execution of sensor calibration or titrant standardization routines. In con-clusion, this automatic workstation con-tributes to the improved understanding of Swedish ecosystems by providing the reli-able measurement of important environ-mental parameters with minimal effort.

Text: Hans-Joachim Muhr, Manager Market Support Group SBU AnaChem

Peter Blomqvist of Umeå saves analysis time with the Rondo30 for pH and conductivity analysis.

www.mt.com/multiparameter-titration

Tips & Tricks

Download everything you need to know about titration

Education is key to understand-ing your titrator and applications. The Basics of Titration guide cov-ers the following areas:

• Theory of Titration• Chemistry and Equations• Automated Titrator Functions• Instrument, Reagent and Sensor Maintenance and Care

Sing up and receive a free copy of the comprehensive booklet.

www.mt.com/titration-basic

3METTLER TOLEDO Academia News 12

Easy Solutionto Ensure Pipette Accuracy

Small pipetting errors can have a big impact on research results. A balance that easily converts for quick

pipette checks, such as METTLER TOLEDO’s NewClassic MS, means the difference between first-time accuracy

and costly rework.

Pipette accuracy checks reduce errors. But they can also be time-consuming and resource intensive. The new convert-ible semi-micro ‘NewClassic MS’ Balances from METTLER TOLEDO combine weigh-ing and pipette accuracy checks in the same easy-to-use instrument. The two-in-one instrument not only provides bet-ter results, but also cost savings.

Easy conversionThe NewClassic MS converts easily into a stand-alone pipette checking station. Firmware guides users through the pi-pette check process. An evaporation trap is the only external device needed to keep

humidity levels stable. This minimizes sample evaporation for accurate results.

Automatic calculation Very small volumes of liquid, such as those dispensed from pipettes, cannot be accurately measured directly. There-fore, ISO 8655-6 recommends gravimetric methods for pipette testing: The test wa-ter dispensed from the pipette is weighed on a balance and the weight value is converted to a volumetric value. In the NewClassic MS, proprietary embedded firmware automatically calculates sys-tematic and random pipetting errors in absolute and relative values. Results are

The balance is quickly converted into a pipette check-station whenever necessary.

An evaporation trap keeps humidity levels stable to minimize evapo-ration and ensure accurate results.

displayed and can be printed out for ref-erence.

Assurance between calibrations Convertible balances, such as the New-Classic MS, are ideal for life science in-dustries where more frequent accuracy checks than the periodic ISO standard calibrations provide added security that pipettes are still delivering exact results.

For more about how METTLER TOLEDO’s NewClassic MS can help eliminate errors and save time with easy pipette accuracy checks, visit www.mt.com/newclassic.

Text: Joanna Imschweiler, Marketing Manager, Classic Balance

www.mt.com/newclassic


Pipe

tting Streamline Effort

for DNA Sequencing Workflow

As an integral part of the International Barcode of Life Project, the

Canadian Center for DNA Barcoding (CCDB) produces a barcode

reference database for standardized gene regions in order to discover

new specimens from plant/animal species. The Liquidator 96 is utilized

in crucial steps along the DNA barcoding process.

The Liquidator 96 is a 96-channel man-ual pipetting system that accommodates high-throughput sample setup. The increased throughput, provided by the simultaneous pipetting of the 96-chan-nels, makes the Liquidator 96 particularly useful for genomics applications, such as

DNA and RNA purification, restriction digestion of DNA, reverse transcription and enzymatic processing of RNA, PCR amplification of DNA templates, ligation of sequences to cDNA and sequencing re-action setup.

DNA Barcoding ProcessDr. Natalia Ivanova, lead DNA scientist at the CCDB, confirms that the Liquida-tor 96 plays an important role in four major steps of the DNA barcoding: DNA extraction from plant/animal tissue, PCR amplification of extracted DNA, cycle se-quencing extension and cycle sequencing cleanup.

Dr. Ivanova uses the Liquidator 96 differ-ently depending on whether DNA is ex-tracted from animal or plant tissue. With animal tissue, the Liquidator 96 is used to deliver lysis buffer, containing protease, to samples. After overnight sample incu-bation, Ivanova’s team uses the Liquida-tor to transfer lysate to a micro-plate for robotic DNA extraction. With plant sam-ples, the Liquidator 96 is used to purify DNA using glass fiber membrane bind-ing. Firstly, plant extract in lysis buffer is transferred from a tube rack to a 96-well plate containing binding buffer using the Liquidator 96. The resulting sample is transferred to a 96-well glass fiber mem-brane purification plate. The Liquidator 96 is then used after sample binding to carry out two wash cycles. After sample drying, the Liquidator 96 is used to pipette


water to the plate, which elutes the DNA from the plate after centrifugation.

Reliable DNA amplification using PCRAfter extraction, DNA is amplified using PCR. Here, the Liquidator 96 is used to de-liver two microliters of purified DNA from the extraction step into a 96-well plate containing PCR mastermix. After sealing and centrifugation, the DNA is amplified using a thermocycler.

After DNA amplification, the PCR prod-ucts are prepared for cycle sequencing us-ing the Liquidator 96. Here, it is used to deliver water to the PCR plate in order to dilute the amplified DNA three-fold. Two microliters of the diluted DNA are deliv-ered by the Liquidator to 96-well plates containing sequencing mastermix. The reaction is then centrifuged and placed

into a thermocycler for DNA extension. This protocol processes 90 sequencing samples in parallel.

After DNA extension, the reactions need to be cleaned up. With the Liquidator 96, the entire volume of the sequencing re-action can be simultaneously transferred to a Sephadex G-50 plate. After centrifu-gation, the samples are transferred with the Liquidator 96 to a plate containing 0.1 mM EDTA. The resulting plate is sealed and placed on a sequence assembly plate prior to loading onto a sequencer for DNA barcode analysis. The final results are imported to the CCDB information man-agement system for accurate tracking.

The protocol above demonstrates the Liquidator 96’s significant value to laboratories requiring accurate high-throughput data collection. In Dr. Ivano-va’s case, the Liquidator 96 most certainly facilitates her workflow.

Text: Rishi Porecha, Ph.D.,Applications Specialist

www.mt.com/liquidator96

Know-how

See how Liquidator 96 processes DNA samples

Follow Dr. Natalia Ivanova,lead DNA scientist, in the 4 short videos to see the fundamental roles that Rainin’s Liquidator 96 plays in processing her DNA samples.

See application videos and down-load the white paper at our CCDB microsite:

www.mt.com/Liquidator-ccdb


Pow

der D

osin

g Secure Handling of Nanoparticleswith Automated Dosing System

The French Alternative Energies and Atomic Energy Commission (CEA)

is active in areas such as low-carbon energies, information technolo-

gies and health technologies. The Laboratoire Capteurs Diamant (LCD)

of CEA, called on Quantos to optimize its nanoparticle suspension

processes in order to guarantee the quality of the diamond deposits it

produces.

It is a little-known fact that diamond is an ‘extreme’ material. Bertrand Bazin, lab manager of LCD in Saclay, France, explains that, "In addition to its hardness, diamond has some surprising properties, espe-cially with regard to thermal conductivity (5 times better than copper), propagation of vibrations (15 times better than water) and electrical insulation (7 times better than glass).”

In its natural state, it is rare, expensive, small, and chemically impure. Happily, LCD has a routine method for synthesizing them: The CVD (Chemical Vapor Deposi-tion) process produces wide film sheets of controlled purity diamonds. This process uses microwaves to excite a carbonated gas

above a silicon sheet on which a suspen-sion of diamond nanoparticles has previ-ously been deposited in order to assist the creation of the diamond film.

The team has acquired recognized exper-tise in nanoparticles but also in numerous other disciplines, such as electrochemistry and materials etc. LCD, therefore, works as much with manufacturers as with other research centers in France and overseas. Mr. Bazin explains, "For example, we have worked on perfecting dosimeters for radio-therapy, new-generation electronic compo-nents and an ultra-sensitive electronic nose (for explosives, toxins and drugs etc.) and, currently, industrial and agribusiness en-vironment monitoring is a boom market.”

Quantos: a vital link in the processIn suspension, the size of certain nanopar-ticles, as well as their concentration and dispersion, changes their properties and especially their toxicity (e.g., metal nanoparticles). "Then we have to control the suspension of nanomaterials, the es-sential element of which is placing them in suspension, splitting the nanoparticle ag-gregates and measuring the size and dis-persion of aggregates/particles," continues Hugues Girard. "Previously, we were using a balance and a volumetric flask to place particles in suspension, even though this initial dosage phase is critical.”

Céline Gesset adds, "Thanks to Quantos, we are now working in complete safety. The QB5-L ensures that we can handle toxic products without risk of exposure and there is no spatula, cupel or cleaning required. In addition, automatic dissolution ensures the repeat accuracy and precision of the concentrations of suspensions used.”

"Several operators, handle the QB5-L on a daily basis, creating and testing up to 30 suspensions a day,” concludes Jacques de Sanoit.

Text: Joël Goursot, Communication Manager, METTLER TOLEDO France

www.mt.com/quantos

Some of the Quantos QB5-L users from LCD (from left to right): Hugues Girard (engineer researcher chemist), Céline Gesset (engineer researcher electronics), Bertrand Bazin (engi-neer researcher co-installation manager) and Jacques de Sanoit (engineer researcher electrochemist).


Monitoring Cell Culture Bio-Processesin Situ and in Real-Time

ReactIR™ and Focused Beam Reflectance Measurement (FBRM®) are proven in situ technologies for real-

time monitoring of principle bioprocess components and changes in cell density. This article highlights the

results of two applications, both demonstrating the value of in situ ReactIR™ and FBRM® in bioprocess

monitoring.

With the increase of biopharmaceutical drugs on the market in recent years, re-search focus has turned to optimizing bio-processes in order to maximize product re-covery and improve product quality though the use of Process Analytical Technologies (PAT). Off-line techniques for monitoring cell cultures are time consuming, may require sample preparation and do not provide real-time bioprocess information, which restricts the ability to control the process. Non-invasive and non-destructive in situ probe-based technologies monitor processes in real-time by providing infor-mation about the bioprocess as it exists in the bioreactor.

Monitoring carbon source and metabolites ReactIR™ is an ATR mid-IR spectro-meter equipped with a DiComp probe. The ability of ReactIR™ to monitor the concentrations of 4 principle analytes,

glucose, L-glutamine, lactose and am-monia in a mammalian cell culture was determined by comparison to offline enzymatic assays (Megazyme, Ireland). The data in figure 1 demonstrates how ReactIR™ was able to monitor component concentrations at both high and low con-centrations.

As cells metabolize glucose and gluta-mine, lactate and ammonia are produced, respectively, meaning there is a strong cor-relation between the metabolically related components. To confirm that ReactIR™ monitors each component independently of the other, further experiments tested the calibration models. In the example shown in figure 2, glucose was spiked into a solu-tion of lactate and ReactIR™ spectra were collected. The data confirms that changes in glucose concentration did not affect the measurement of lactate even when the sample was spiked with a 30 mM of glucose.

Monitoring cell densityThe in situ monitoring of cell density changes was proven using FBRM® which counts cells and measures cell dimensions. Cell counts determined by FBRM® dem-onstrated a strong linear relationship with cell density with a coefficient of determina-tion greater than 0.99 (figure 3).

ConclusionsThe non-invasive and non-destructive in situ, real-time monitoring of principle components and changes in cell density, as they exist in the reactor, is possible with ReactIR™ and FBRM®, respectively. ReactIR™ and FBRM® eliminate the need for sampling and sample workup, while reducing the risk of contamination. The real-time information provided by ReactIR™ and FBRM® enables bioprocess understanding and immediate process control.

Text: B. Rice, University College Dublin.

www.mt.com/ac-biotech

Figure 1. Actual and predicted values for 4 principle components of a cell culture.

Figure 2. Changing glucose concentration does not affect the lactate concentration as measured by ReactIR™.

Figure 3. Effect of cell density on total cell count as measured by FBRM®.

Tota

l Cou

nts

(#/s

)

Cell Density (cells/mL)

m = 3.22x10-5

R2 = 0.994

m = 1.36x10-5

R2 = 0.999

Total Counts 1

Total Counts 2

350

300

250

200

150

100

50

02e+60 4e+6 6e+6 8e+6 1e+7


Goo

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Prac

tices

GWP® – Good Weighing Practice™

Guaranteed better weighing results

GWP® helps ensuring consistent product quality and avoiding out-of-spec results or bad production batches. With GWP® you comply with all regulations relevant to your industry for a minimal time and financial investment.

GWP® provides documentation to verify the accuracy of your balances and recommends optimal testing procedures.

www.mt.com/gwp

GTP® – Good Titration Practice™

Reliable results and dependable titration in practice

GTP® forms the basis of depend-able and risk-free titration. The appropriate titration system is selected based on user require-ments, and professionally commis-sioned and installed.

GTP® helps protecting your invest-ment far into the future, support-ing you with tailored training and equipment qualification. It also pro-poses schedules for regular inspec-tion and preventive maintenance.

www.mt.com/gtp

GPP™ – Good Pipetting Practice™

Optimize workflows for more reliable and repeatable results

Improve your data quality with GPP™ – our comprehensive, sys-tematic approach to maximizing pipetting accuracy and repeat-ability.

GPP™ is grounded in 40+ years of expertise working side-by-side with researchers in order to achieve the highest level of accuracy and preci-sion across all applications.

www.mt.com/gpp

Good Measuring Practices by METTLER TOLEDO is a global program supporting you in laboratory

and production environments with quality assurance recommendations for weighing, pipetting and

chemical analysis. The five steps of every Good Measuring Practices guideline cover the entire life-

cycle of your instruments. By focusing on your processes and its associated requirements and risks,

we provide you with effective measures to maximize operational security and minimize testing

efforts and process risk.

Good Measuring PracticesReliability Throughout the Lifecycle


Risk CheckAll of the Good Measuring Practices guidelines start with an evaluation of your measurement processes and an assessment of their associated risks. Based on this information, you are provided with straightforward recommendations for selecting, installing, calibrating and operating your devices.

GoodMeasuringPractices

1Evaluation

2Selection

3Installation /

Training

5Routine

Operation

4Calibration /

Qualification

Check your risks in weighing, titration and chemical analysis

For more Information:Good Measuring Practices www.mt.com/gp

Good Measuring PracticesReliability Throughout the Lifecycle

GDRP™ – Good Density and Refractometry Practice™

Minimize risk and improve performance

GDRP™ is based on our expertise in density and refractive index measurement. Understanding the reasons and sources of measure-ment errors helps improving qual-ity while reducing risks and costs.

Professional commissioning, train-ing, qualification and seamless documentation guarantee compli-ance with your process require-ments from the very first step.

www.mt.com/gdrp

GEP™ – Good Electrochemistry Practice™

Optimized pH results from the beginning

GEP™ assists with professional tools and documentation to get reli-able results for your laboratory pH, conductivity, DO, ion and redox applications.

Use GEP™ to select the correct calibration solutions to comply with regulations, perfectly maintain your sensors to optimize product lifes-pan and reduce costly measure-ment mistakes to a minimum.

www.mt.com/gep


Prod

uct H

ighl

ight Simple and Flexible Solution

for Dynamic Mechanical Analysis

Dynamic Mechanical Analysis (DMA) is the method

used to measure the mechanical and viscoelastic prop-

erties of materials as a function of temperature, time

and frequency when subjected to periodic stress under

a defined environment. The analyzed materials encom-

pass thermoplasts, thermosets, composites, elasto-

mers, ceramics and metals. In addition to operation in

DMA mode, the DMA 1 permits measurements in TMA

mode and under various environments.

Controlled humidity experimentsThe humidity generator and humidity chamber allow samples to be analyzed under varying relative humidity condi-tions at a given temperature.

Immersion measurementsThe fluid bath option permits the user to perform immersion studies on a sample. All DMA modes are supported by the immersion option.

TMA experimentsIn addition to the established DMA modes, the DMA 1 permits operations to be performed under constant force (TMA or creep), in stress-strain or stress relaxation mode.

www.mt.com/dma_1


Reliable and Intuitive pH Measurementwith New SevenCompact™

For research groups and laboratory

courses alike, the intuitive operation

and versatility of SevenCompact

series fulfil the needs of lab

researchers and students.

Perfect ergonomics with uPlace™

The electrode arm can be operated with one hand, moving straight up and down to the position where it works best for your sample.

Broad sensor offeringFind the best sensor for your application with tips and tricks from:www.mt.com/electrode-guide.

One single keypressBy switching to uFocus™ view, you will not be distracted by information of less importance.

www.mt.com/sevencompact

www.mt.com/academiaFor more information

Mettler-Toledo AGLaboratory DivisionIm LangacherCH-8606 Greifensee, Switzerland

Everything in Our Lab Works With Just One Click®

The One Click operation provides unsurpassed user guidance, secure navigation and easily selectable and adaptable methods.

Learn more and see One Click in action: www.mt.com/one-click-titration

Bob, production assistant: "All I have to do to analyze the intermediate product every hour is press a Shortcut."

Joanne, laboratory assistant: "Thanks to my personalized Homescreen, I can perform all the different analyses very quickly and efficiently."

Clark, quality assurance: "The individual user rights feature means that I can make sure that users stick to our quality guidelines."

Know one – know allOne Click is a common interface for all your lab instruments. This means, if you can program and run analyses on a balance, you can also do it on a titrator, without extra information required.

See only what is relevant to youUser screen individualization per-mits you to adapt your home screen to your requirements: You only see what you need and avoid confusion by unneeded functionalities.

Play by the rulesAdherence to SOPs is now a func-tionality of your instrument via the pre-programming of methods. Transcription and calculation errors a thing of the past.

Key benefits of One Click operation:

academia 12 weighing and analysis in the laboratory news

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