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Fall 2002 / International Edition Rapid LCMS Analysis of Common Pesticides/ Herbicides Shimadzu's LCMS-2010 revolutionizes method development approach 5 New Products Shimadzu unveils SIL-HT/ELSD-LT 8 Real World Solutions SIL-HTc Competitive Carryover Performance 11 Compliance Corner Compliance Software 14 Automation Solutions via Strategic Alliances Therapy for the Pharmaceutical Industry Automation Solutions via Strategic Alliances Therapy for the Pharmaceutical Industry Rapid LCMS Analysis of Common Pesticides/ Herbicides Shimadzu's LCMS-2010 revolutionizes method development approach 5 New Products Shimadzu unveils SIL-HT/ELSD-LT 8 Real World Solutions SIL-HTc Competitive Carryover Performance 11 Compliance Corner Compliance Software 14 C190-E081

Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

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Page 1: Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

Fall 2002 / International Edition

Rapid LCMS Analysis of Common Pesticides/HerbicidesShimadzu's LCMS-2010 revolutionizes method development approach 5

New ProductsShimadzu unveils SIL-HT/ELSD-LT 8

Real World SolutionsSIL-HTc Competitive Carryover Performance 11

Compliance CornerCompliance Software 14

Automation Solutions via Strategic Alliances Therapy for the Pharmaceutical Industry

Automation Solutions via Strategic Alliances Therapy for the Pharmaceutical Industry

Rapid LCMS Analysis of Common Pesticides/HerbicidesShimadzu's LCMS-2010 revolutionizesmethod development approach 5

New ProductsShimadzu unveils SIL-HT/ELSD-LT 8

Real World SolutionsSIL-HTc Competitive Carryover Performance 11

Compliance CornerCompliance Software 14

C190-E081

Page 2: Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

Automation SolutionsviaStrategicAlliancesTherapy for the Pharmaceutical Industry

“Make this faster, more reliable and flexible?”The request to automate, speed analysis andpurification of compounds in the areas ofdrug discovery and development has beenthe mandate issued to analytical instrumentcompanies!

The message is clear: if you watch…it willhappen without you and your company.”

C o v e r S t o r y

Rapid technological advancesin combinatorial chemistry,genomics and proteomics

have dramatically altered drug dis-covery and development during thepast twenty years. The pace in thepharmaceutical industry and lifesciences marketplace has escalatedfrom a comfortable canter to an allout sprint in order to deliver drug-to-market. In turn, this has driventhe technology industry beyondimagination to automate highthroughput sample preparation andanalysis. The driving engine being:“How can you make this faster,more reliable and flexible?” Therequest to automate, speed analysisand purification of compounds inthe areas of drug discovery anddevelopment has been the mandateissued to analytical instrumentcompanies! The message is clear: ifyou watch…it…will happen with-out… you and your company. Thatis “old news," but one disciplinedefines the other’s destiny.Automate now or perish!

Successful and competitive deliveryof drug-to-market requires stream-lined high throughput analyticalprocedures and, therefore, automa-tion when and wherever possible inboth drug discovery and develop-ment. The pace at which pharma-ceutical companies are moving has put an extreme demand onanalytical instrument companies todevelop faster, more robust instru-ments to address the exponentiallyincreasing amount of samples thatneed to be analyzed. To developthe necessary equipment to satisfythe immediate market demand ofhigh throughput is a challengeinvolving software and hardware.Instrument companies must weighthe risk of producing a new productto address the current market

2 Fall 2002 www.shimadzu.com

Page 3: Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

demand against whether or notthat instrument will spontaneouslybecome obsolete due to the rapidlyevolving marketplace. However,there are several options to thisdilemma:

1. Take a slightly more conservativeapproach in manufacturing amore “mainstream” product.

2. Form strategic alliances or part-nerships with pharmaceuticalcompanies to enhance or modifyexisting hardware or software.

3. Combine technologies availablefrom your own company andthird parties into an IntegratedSolution targeted to the needs ofa specific market segment.

4. Provide “virtual instrument”plug-ins so others can controlyour product.

In the pharmaceutical market seg-ment all options find their way intothe proverbial think tank and areoften times not mutually exclusive.However, the second option offorming strategic alliances is onethat has suited Shimadzu to moveswiftly and strategically in cadencewith the radically changing phar-maceutical market. It is a boldmove for a conservative company,but the strategy has yielded newproduct innovation and technologiesthat would not have been realizedoperating individually. Shimadzuhas been pioneering strategicalliances, providing almost immedi-ate automation technologies usingan existing state-of-the-art HPLCproduct line.

Cutting-Edge Technologiesvia Key Strategic Alliances:A Brief History

The key to success for any companyis to excel in producing a qualityproduct and good customer service.

How is that accomplished in therapidly evolving pharmaceuticalmarket? Dynamic strategic visionand leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic allianceswith key customers. Through thesetypes of alliances the pharmaceuticalindustry and Shimadzu have beenable to work closely together withthe customer/collaborator toaddress immediate demands andexpand upon the functionality ofexisting equipment. Fortunately,Shimadzu HPLC equipment,

through its open architecture,inherently affords this flexibility.This provides the opportunity tomodify or enhance existing func-tionality and the ability to plug intoequipment from third-party vendors.This also opens the door forresearchers to explore the fullpotential of the equipment and setsthe stage for strategic alchemy withShimadzu. Shimadzu MarketingCenter’s pursuit of these types ofalliances was initially and intri-cately involved with Dr. Harold

3Summer 2002 www.shimadzu.com

Discovery VPTM Preparative System

Discovery VPTM Analytical System

3Fall 2002 www.shimadzu.com

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4 Fall 2002 www.shimadzu.com

Weller at Bristol-Myers Squibb(BMS). Dr. Weller has been and isactively pioneering efforts to auto-mate all aspects of sample handling,analysis and purification in theEarly Lead Discovery group at BMS.

New Enabling Tools For DrugDiscovery—Discovery VP

This collaboration has resulted indeveloping new enabling tools fordrug discovery: namely theDiscovery VP™ Analytical andPreparative fully automated HPLCsystems. The Discovery VP systemswere described in detail in LCWorld Talk Volume 5, Number 1,April 2000. Discovery VP was oneof the early and complete walk-upautomation chromatography systemsthat addressed: multiple columnselection, scalable multiplexing ofHPLC components, microtiterplates, UV and photodiode arraydetection, fraction collection, intuitive user interface with e-mailnotification to both user andsystem administrator.

Basically, the Discovery VP systemshave successfully addressed most of the demands for automated analytical and preparative HPLCanalyses with easy to use queue-

based control software and multi-plexing capability. The intuitiveinterface and minimal requiredparameters enable users to becomeproficient with little training. Theeasily mastered software operatesthe entire system, so the laboratorychemist need not spend time learn-ing how to properly operate andcontrol HPLC equipment, optimizemethodology, or adjust data acqui-sition conditions. Essentially, theoriginal idea behind these auto-mated systems is to take the chro-matography out of the chemistry,allowing the combinatorial chemistto spend more time performingsyntheses. The original idea andtechnology have evolved over thepast few years.

New Evolving Technologies:Automation Enhancement

From Discovery VP to a NewBroad Spectrum AutomatedHPLC/MS System

Shimadzu has always endeavoredto provide technology to key targetmarkets and once again providesnew software and hardwareenhancements in our new auto-mated LCMS system. Built upon

the featured requisite demandsfrom the major pharmaceuticalmarket already contained in theDiscovery VP, Shimadzu’s enhancedautomated LCMS system nowemploys the highly sensitiveShimadzu LCMS-2010 MassSpectrometer (below) in addition toits predecessor the QP8000 MS.

Conclusion

The applications and automatedchromatography tools presentedhere may seem intuitively simple,but the implementation or puttingthe technology in place is not a trivial matter. The genesis of thesecutting-edge technologies was madepossible through key strategicalliances with Dr. Weller and BMSand would not have been possiblewithout this collaborative effort.Shimadzu and Bristol Myers Squibbare successful companies that havejoined in scientific excellence toattain therapeutically useful goals.Strong strategic alliances with keypeople in both the pharmaceuticalindustry and Shimadzu haveafforded the opportunity to stay onthe cutting edge while adding valueto customer demand.

New Evolving Technologies

Feature Enhancements of Shimadzu’s new fully automated LCMS System in addition to those of Discovery VP include:

■ New and more robust 32 Bit Chromatography AutomationSoftware Package

■ Shimadzu’s LCMS-2010 High Sensitivity Mass Spectrometer ■ Mass-Directed Fraction Collection (prep. system)■ Multiple mass targeting■ Highly Accurate Ratio Sample Injection■ Dual wavelength UV detection■ Up to 10 Column Switching (analytical system)■ Reinjection capability■ Reproducible flow splitting capability

LCMS-2010

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The importance of rapid and reliable determination of pesticides

and herbicides in health, nutrition, and environmental studies

cannot be overstated. The use of Liquid Chromatography/Mass

Spectrometry (LCMS) offers some distinct advantages for such mea-

surements, including the capability of direct measurement of pesticides

and herbicides without the need for derivatization. Productivity is

thereby increased as a result of higher sample throughput.

In this communication, we report on rapid LCMS analysis techniques

for certain herbicides and pesticides using the Shimadzu LCMS-2010

and demonstrate enhanced productivity, increased capabilities during

data acquisition, and reductions in total run time.

Shorter run times are used when employing fast gradient techniques

and baseline separation is no longer needed for most LCMS applications.

LC coupled with MS increases the selectivity and sensitivity of

traditional HPLC methods. Shimadzu’s new LCMS-2010 offers a

highly sensitive Single Quadrupole Mass Spectrometer with improved

throughput, allowing end users to dramatically increase their

productivity.

A N A L Y S I S S E C T I O N

Rapid LCMS Analysis ofCommon Pesticides/Herbicides

“The Shimadzu LCMS-2010

can be an effective analytical

tool for developing robust

qualitative and quantitative

analyses for pesticides and

herbicides. The versatility

built into LCMS Solution®

software streamlines the

time and effort spent on

developing these methods.”

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Gradient for separation of pesticide mixture

TIME(min)

%B

The robustness of the method isindicated by excellent retentiontime repeatability for the test com-pounds, as indicated in Table 1.

Table 1. Retention Time Repeatability

Compound RetentionTime RSD

Dimethoate 0.5%

Atrazine 0.2%

Isoproturon 0.2%

Prometryne 0.3%

Chloroxuron 0.5%

Analysis Sensitivity was not compromised,as indicated by LOD's of at least1ng/mL (2 pg on-column) for all of the compounds. The minimumS/N was 5 in the case ofDimethoate, while the remainingcomponents in the test mixturehad S/N values ≥10.

These data are shown in Table 2.The figure on the following pagedisplays the data on the sensitivitydetermination for Prometryne.

Conditions

Instrumentation:Shimadzu LC-10ADvp PumpsShimadzu SIL-HT AutosamplerShimadzu CTO-10ASvp ColumnOven

Conditions:Flow Rate: 0.700 ml/minInjection Volume: 2 µlColumn: ES Industries PremierC18 2.1cm x 50mm, 5 micronsMixer Volume: 100 µlColumn Temperature: 60°C

Mobile Phase:A: Water (0.1% Formic Acid, v/v)B: Methanol (0.1% Formic Acid, v/v)

Materials and Methods

The pesticide standards:Dimethoate, Atrazine, Isoproturon,Prometryne and Chloroxuron wereobtained from Chem Service, WestChester, PA. Methanol wasobtained from Burdick and Jack-son Inc., Muskegon, MI. TOCgrade water was used for all experiments. The analytical LCMScolumn was obtained from ESIndustries.

Structures of pesticide and herbicide standards

LCMS Conditions: Shimadzu LCMS-2010Mode: Positive Block Heater: 250°CAPCI Probe: 450°C CDL Heater: 250°C

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Results and Discussion

Mass Spectra of Standards

Separation of Standards

Table 2. Signal-to-Noise Values

Compound Conc. S/N

Dimethoate 1 ng/ml 5:1

Atrazine 1 ng/ml 24:1

Isoproturon 1 ng/ml 11:1

Prometryne 1 ng/ml 18:1

Chloroxuron 1 ng/ml 28:1

Conclusion

The Shimadzu LCMS-2010 can bean effective analytical tool fordeveloping robust qualitative andquantitative analyses for pesticidesand herbicides. The versatility builtinto LCMS Solution® softwarestreamlines the time and effortspent on developing these methods.Overall throughput increases, butwithout compromising dataintegrity. The Shimadzu LCMS-2010 can revolutionize methoddevelopment approach, reachinglow levels of detection unmatchedby any other single-quad MS.

Page 8: Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

In any laboratory today, thesingle common denominator isthe need for multi-tasking in

equipment as well as in staff. Goneare the days when a single instru-ment operated by a single expertcan be dedicated to a single task.This is especially true for laborato-ries engaged in liquid chromatogra-phy techniques. The maturation ofHigh Performance LiquidChromatography (HPLC) as ananalytical discipline has dictatedthat the systems be easily operatedby operators who are not necessar-ily experts in chromatography. Anadditional requirement is that theequipment possesses the maximumcapacity for self-validation andreporting. There can be little doubtthat mass detection is becomingmore and more important inHPLC, and there is always the

critical need for increased samplecapacity, high sample throughput,maximum data repeatability andaccuracy, and minimum samplecarryover. As the needs of the analyst can change rapidly, thecross compatibility of hardwareand software becomes more andmore important.

The requirements described aboveare daunting, but Shimadzu hascontinued its tradition of excellencein HPLC with the introductionrecently of three outstanding prod-ucts. Shown in Figure 1 is the SIL-HT high performance stand-aloneautosampler. The unit is availablein two models, the SIL-HTa andthe SIL-HTc with sample cooling.The SIL-HT enhances analysisthroughput by fast sample injectionand high sample capacity. Injectionspeed is 18 seconds for injecting

samples of 10µL (1/2 of SIL-10ADVP).Quite simply, this autosampler isthe finest available and hasunmatched sample throughput,data repeatability (Figure 2), andlinearity (Figure 3). Recent evalua-tion of the SIL-HT by pharmaceuti-cal companies has met with a mostfavorable response, as have similarevaluations by contract laboratoriessuch as Covance. All of these enter-prises require the highest perfor-mance and versatility in anautosampler for LC and LCMSapplications, including using theSIL-HT as an autosampler fortriple-quad MS.

Seeing is believing, so the sayinggoes, and the data on repeatabilityand carryover for the SIL-HT isreally worth seeing: 100 injectionsof 0.05 mg/mL caffeine, 10µL each,no rinsing, %RSD=0.230.

NEW PRODUCTSNew Liquid Chromatography productsfrom Shimadzu find wide acceptance in many laboratory settings

Figure 1—The SIL-HT Autosampler

• SIL-HT• ELSD-LT

8 Fall 2002 www.shimadzu.com

Autosampler enhances analysis throughput by fastsample injection and high sample capacity

SIL-HT AUTOSAMPLER

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9Fall 2002 www.shimadzu.com

high-performance measuring pump(6nL step resolution) for enhancedaccuracy. This results in superiorrepeatability over conventionalautosamplers. In addition, precioussamples are not wasted since thetotal volume aspirated is injected.

The SIL-HT also has excellentsample capacity:

350 x 1mL vials (SIL-HTa/c)

210 x 1.5mL vials (SIL-HTa) and140 x 1.5mL vials (SIL-HTc)

Carryover in the SIL-HT is guaran-teed to be less than 0.01%,unmatched among commerciallyavailable LC autosamplers. In thetest results shown in Figure 3, therewas no detectable carryover inblank injections following 10repeat injections of naphthalene.

Conditions

Flow rate: 1 mL/minMobile phase: methanol/water

(60/40 v/v)Wavelength: 254 nm

To minimize carryover, the outersurface of the sampling needle ofthe SIL-HT’s autosampler is coatedusing a special, innovative surfaceprocessing technology (patentpending). Furthermore, by employ-ing newly developed rotor seals andneedle seals made of PEEK, theSIL-HT dramatically reduces contamination, even with the mosthighly adsorbent sample com-pounds. As a result, the SIL-HTdemonstrates nearly zero samplecarryover.

The SIL-HT uses a needle-in-the-flow-path injection design (alsocalled a direct injection method ortotal volume injection method) and

100 x 4mL vials (SIL-HTa/c)

4 x MTP (1536 samples) (SIL-HTa/c)

96-well Standard and Deep Well

384-well Standard.

The high sample capacity, high-speed injection capability, injectionvolume repeatability, and ultra-lowcarryover make the SIL-HT anunsurpassed performer in high performance LC autosamplers.

Conditions: SIL-HTc LC-10ADvp, SPD-10AV VP-ODS column 4.6 x 150 mm60%MeOH:H2O/No Rinse0.05 mg/mL caffeine, 254 nmSample Injection Speed = 3 µL/s 5 X 1, 5, 10, 25, 50, and 100 µL

SIL-HT InjectionVolume Linearity

Figure 2

Figure 3—Injection Repeatability Carryover of Naphthalene in the SIL-HT

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10 Fall 2002 www.shimadzu.com

Shimadzu has now completed the family of HPLC detec-tors with the introduction of the ELSD-LT. The ELSD-LT is ideally suited for applications in drug discovery,

natural products development, combinatorial chemistry, andfood and beverage industries. Analytes are detected regardlessof their chromophoric properties. Essentially, all compoundsare detected even if they do not contain a chromophore or anelectroactive group.

Shimadzu’s ELSD-LT low-temperature evaporative light scatter-ing detector transforms a target compound to particulate formby evaporating the mobile phase, and then measures the lightscattered by these particles to detect the compound. In general,the detector will deliver a signal for all compounds that do notevaporate or decompose during the mobile phase evaporationstage. Further, the detector produces stable baselines during gradient elution chromatography as its response is independentof the bulk spectral properties of the eluent. The ELSD-LT produces nearly uniform detection sensitivity for the majority of analytes, regardless of their physical properties. The low-tem-perature integrated design prevents the decomposition of ther-mally labile compounds, resulting in increased assay sensitivity.

The ELSD-LT is a universal mass sensitive detector with thefollowing key benefits:

■ Low temperature operation–The ELSD-LT optimizes sensitivity of thermally labile com-pounds by low-temperature evaporation of the mobile phaseusing a three-step process. This system can evaporate a mobilephase consisting of 100% H2O at 32˚C. Other systems requirea special accessory to evaporate the mobile phase. ELSD-LThas an integrated design for optimizing sensitivity, performanceand ease of operation.

■ New Cell Design Reduces Band Broadening–The innovative cell design minimizes band broadening providing a similar bandwidth as UV, UV-VIS detectors.

■ Excellent Performance using Gradients–The ELSD-LT provides excellent resolution of analytes sincegradients can be used to optimize separations. The mobilephase is removed from the eluent even at high flow rates suchas 4ml/min. with uncompromised resolution.

■ Quantitation of all Analytes in a Sample–Essentially all compounds in a sample provide a signalallowing accurate quantitation. The detector response isrelated to the mass of the compound, not UV absorbance,making this detector extremely useful for the combinatorialchemist. The ELSD-LT is a universal detector.

Shimadzu Introduces the Low-TemperatureEvaporative Light Scattering Detector

ELSD-LT

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11Fall 2002 www.shimadzu.com

SIL-HTc High Throughput Autosampler

DVANCES IN THE FIELD OF DRUG DISCOVERYhave resulted in a tremendous production of potentially thera-peutic compounds and, thus, another “bottleneck” downstreamfrom synthesis to drug development. Candidate compounds

need to be rapidly analyzed for structure elucidation and evaluated forefficacy and toxicity. Bioanalysis is now perhaps the most prominentactivity in drug development and the preferred tools for these types ofanalyses are High Performance Liquid Chromatography (HPLC) coupledwith triple quadrupole mass spectrometry (LC-MS/MS). Successful devel-opment of accurate and dependable bioanalytical methods is thereforeextremely critical. Method development to determine levels of drugs andtheir metabolites in biological fluids can present an array of analyticalproblems that can interfere with the assay. The causes of assay interfer-ence are often attributed to sample carryover, particularly with “sticky”compounds having an affinity for the flow-path composition and/oruncleared micro dead volumes in the HPLC system and, in particular, theautosampler. In addition, the increased sensitivity of triple quadrupolemass spectrometry tends to magnify the detection of carryover. Carryoverhas been conventionally reported using UV detection. Mass spectrometrywas used in this collaborative study with Covance Laboratories (Madison,WI, USA) as a more sensitive determination of sample carryover using thenew high throughput SIL-HTc autosampler. Carryover performance ofthe SIL-HTc was compared to that of other commercially available HPLCautosamplers on the market.

SIL-HTc COMPETITIVE CARRYOVER PERFORMANCE ANALYZED BY LC-MS/MS: A Collaborative Study between Shimadzu Marketing Center* and Covance Laboratories**

REAL WORLD

S.M. Wishnies*, M. Nishimura*,

T. Ueda*, N. Tatsumi*, Y. Maeda*,

S. Maruyama*, A. Ueyanagi*,

T.D.J. Halls**, S. Zhou** and X. Jiang**

(Shimadzu Corp., Kyoto, Japan* and CovanceLaboratories, Inc., Madison, WI, USA**)

A

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12 Fall 2002 www.shimadzu.com

E X P E R I M E N TA L

LC-MS/MS Components:

(1) SIL-HTc High ThroughputAutosampler with CoolingCapability and built-in systemcontroller (below).

(2) Commercially Available HPLCAutosamplers denoted as: A, B,C and D.

All autosamplers were configuredwith the same two Shimadzu LC-10ADvp analytical pumps (highpressure mixing) and MS/MS (SciexAPI 365) for direct comparison ofcarryover performance.

LC-MS/MS Conditions

The Shimadzu LC system was configured with the SIL-HTc andthe commercially availableautosamplers (A, B, C or D) andcoupled with a Sciex API 365 triplequadrupole mass spectrometeroperated in positive turbo ionspraymode. A typical C18 (50 x 4.6mm,3.5µm particle size) column wasemployed; flow rate: 0.400mL/ min.;column temperature: 40˚C; injectionvolume: 20µL.

The mobile phase consisted of: (a) 5mM ammonium formate in 0.1%formic acid: 0.1% formic acid in acetonitrile (55:45, v:v). The massspectrometer was operated in positiveionspray mode. MRM ions forCompound X: 386.3/167.1; dwelltime: 300 msec; capillary voltage: 2.5kV; source temperature: 400˚C; colli-sion gas: N2; collision energy: 37V.

AUTOSAMPLER % CARRYOVER

A 0.0129

B 0.00924

C 0.0170

D 0.0144

Shimadzu SIL-HTc 0.00572

RESULTS

According to the results in Table 1, the SIL-HTc autosampler outperformed orgave the least amount of measurable carryover (~ 2 to 3 times less) compared to theother autosamplers tested (see FIGURE 1). As mentioned in Note 2, extensive rinsing,multiple rinse solvents (when applicable) and/or both were used for the otherautosamplers. Extensive rinsing correlates with increased cycle time. The SIL-HTc wastested using the default injection routine consisting of a 20-sec injection cycle timewithout employing extensive rinsing.

Table 1. Percent Carryover Comparison of the Commercial Autosamplers and the SIL-HTc.

FIGURE 1: TYPICAL CHROMATOGRAM of SIL-HTc CARRY OVER

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C O N C L U S I O N

Sample carryover has progressivelybecome more of a problem in drugdevelopment, especially in the areaof bioanalysis, for a number of reasons. Given that carryover isdependent upon compound structureand thus the resulting chemicalproperties of that structure in amultitude of different environments,there are other obvious variables toconsider. The sample matrix orvehicle is critical when developingan accurate and quantitative assay.The compound and its metabolitesare most likely in biological fluidssuch as blood, urine plasma and/orserum to name a few, which furthercomplicates assay developmentbecause of the very nature of thesefluids (proteins, salts and otherinterfering components).

The compound/metabolites thenusually need to be extracted beforereconstituting into an appropriatesolvent or buffer for LC-MS/MSanalyses. Providing that most ofthese problems have beenaddressed, the analytical equipmentis then suspect and the carryoverseems to be magnified by employingthe increased sensitivity of a massspectrometer as the detector. Theautosampler would then be themost logical component of theHPLC system contributing tosample carryover.

Shimadzu has approached the carryover problem with a newlydesigned high throughput autosam-pler, the SIL-HTc. The rapid cycletime does not sacrifice throughput

and the extremely low carryovermakes it the perfect autosamplerfor these types of LC-MS/MS applications and others. It wasclear that the SIL-HTc had theleast amount of carryover in thisstudy. Shimadzu incorporated sev-eral new design features in the SIL-HTc to accomplish this. The newlyengineered SIL-HTc developmentfeatures include:

A Special metallic coating of thesample needle

B Use of a PEEK™ rotor seal

C Modification of the flow paths

As a result, the new SIL-HTc hadthe least amount of sample carry-over, minimal rinsing with a rapidcycle time of 20 seconds.

A C K N O W L E D G E M E N T S

Shimadzu Marketing Center(Shimadzu Corporation) would liketo acknowledge and extend greatthanks to Timothy D.J. Halls,Ph.D. (Vice President ofPharmaceutical Chemistry) atCovance Laboratories Inc.Madison, WI for allowingShimadzu to collaborate withCovance in this study at CovanceLabs, and for the expertise ofShaolin Zhou, Ph.D. and XiangyuJiang, Ph.D. for performing theexperiments and data analyses intheir labs at Covance. We deeplyappreciate your efforts.

REAL WORLDS A M P L E P R E PA R AT I O N

Sample Compound X (CovanceLaboratories, Madison, WI, USA)was used as the model compoundfor testing the carryover of variousHPLC autosamplers. Sample solu-tions were prepared by diluting thestock standard solution ofCompound X (1.00 mg/mL inmethanol) with methanol/water(1:1,v/v). The tested samplesincluded high standard (1.00µg/mL), low standard (0.500ng/mL) and blank solvent (5 mMammonium formate: acetonitrile:formic acid, 60:40:0.1 v: v: v).

Test Procedure

For each test, 20 µL of a blanksample was injected into the LC-MS/MS immediately after ahigh calibration standard. The carryover (%) was calculated as therelative peak area of the blankagainst that of the high calibrationstandard.

Note 1: The sample for the studywas proprietary and thereforestructural information was notavailable; however, the sample wasdescribed to be basic and veryhydrophobic

Note 2: The rinse protocols for the other autosamplers may haveincluded extensive rinsing with one or more solvents to eliminatecarryover. The SIL-HTc was usedin default, 20-second cycle timewithout extensive rinsing.

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C O M P L I A N C E C O R N E R

Compliance Corner is a new section to LC World Talk that will be dedicated to updates and activities related to

regulatory issues. The editor also welcomes questions to be addressed in the next issue. Please e-mail questions to

[email protected]. Please visit our Web site at www.shimadzu.com.

Shimadzu Offers a Software Suite toAssist Customer Compliance to Title 21, Code of Federal Regulations, Part 11

IT IS NOW CRUNCH TIME for the pharma-ceutical industry, CROs, and biotech busi-nesses alike to consult quality standards, (ISO,

EN, TICKIT, GMP, GLP, GCP, ANSI, ASTM,GALP), streamline their processes, perform GAPanalyses and remediation to prepare for thatinevitable FDA audit. Once your company filesclaim to be 21 CFR Part 11 to the FDA, the flag isup. Failure to comply can be quite drastic and canresult in the FDA issuing a warning letter, pullingproduct from market, astronomical fines, criminalinvestigation and/or closing down the shop. Sincethe introduction of 21 CFR Part 11 (ElectricRecords and Signatures) in 1997, industry has hadtime to perform GAP analyses and explore ways toeffectively execute plans to comply.

During this time Shimadzu Corporation has pro-vided global seminars on regulatory issues involv-ing 21 CFR Part 11 and how Shimadzu as avendor provides the tools necessary to assist customer compliance, from hardware to softwaresolutions. Before introducing the suite of enablingcompliance technologies, we have asked attendeessome critical questions that are essential in build-ing an effective compliance initiative:

Has a company-wide validation plan been devisedto identify all systems that are required to be validated and has this list been prioritized? Havepersonnel been trained and has a timeline been

Shimadzu Corporation’s Dr.Hayakawa (ShimadzuLiquid ChromatographyBusiness Unit) presentedwork on Shimadzu’s state-of-the-art compliant dataarchiving software package,CLASS-Agent.

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15Fall 2002 www.shimadzu.com

established to realistically implement these goals?

Has necessary software andcomputer systems been validatedaccording to internal and FDAguidelines such that they complywith the recommended steps forregulatory compliance - testingand validating computer hard-ware, software, and analyticalsystem configurations whichacquire, store and manage critical data?

What vendor can provide youwith the necessary tools to bestsuit your needs and can they beused to build validation into thesystem acquisition process?

So the question remains: haveyou selected the best vendor toassist regulatory compliance? Forthe past 3 to 5 years, Shimadzuhas been taking part as a vendorto go aboard to help customersunderstand 21 CFR Part 11through a series of seminarssponsored by Shimadzu or co-sponsored with other vendorsand organizations such as theInstitute of Validation Technolo-gies (IVT). Just recently, Shimadzuco-sponsored an IVT seminar onRegulatory Compliance in Tokyo,Japan to introduce Shimadzu’ssuite of software compliancetools: Lab Solutions, CLASS-VPand CLASS-Agent for dataacquisition, storage, archiving,and audit trail, that comply with21 CFR Part 11. Some vendorsmarket compliance, but Shimadzuhas painstakingly addressed allpoints of 21 CFR Part 11 and is

confident these software toolsenable the end user to comply.

Lab Solutions, CLASS-VP andCLASS-Agent comprise the Shimadzu regulatory toolbox toassist end users on the road to

compliance. Compliance with 21CFR Part 11 is achieved throughthe integrated data managementsystem for all common labora-tory instruments. Instrumentsincluded in this suite management system are LC, GC,GC-MS, LC-MS, UV, FTIR, AAand balances. CLASS-Agent provides electronic record andelectronic signature with securedatabase archiving that isrequired for compliance. In addi-tion, a network-based browser isprovided for review and autho-rization of data packages includ-ing raw data, all processedresults files, meta files anddescriptive notes.

As a global vendor of Chromatog-raphy Instruments, Data and Data Archiving Software Systems, Shimadzu can assist you withyour regulatory needs.

SHIMADZU exhibit at IVT seminar in Tokyo, Japan.

“For the past 3 to 5 years,

Shimadzu has been taking

part as a vendor to go aboard

to help customers understand

21 CFR Part 11 through a

series of seminars sponsored

by Shimadzu or co-sponsored

with other vendors and

organizations...”

Page 16: Automation Solutions via Strategic Alliances · market? Dynamic strategic vision and leadership are critical ingredi-ents to success, but equally impor-tant is forming strategic alliances

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