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THE BIOCHROM 30+AMINO ACID ANALYSIS SYSTEMThe gold standard dedicated Amino Acid Analyzerfor in vitro diagnostic use and research

"Our Biochrom amino acid analyzer was installed in2004, and we have been using analyzers from Biochromfor at least 10 years for patient and animal samples.We are a relatively busy laboratory and typically runapproximately 1000 samples per year. We have beenpleased with the performance since installation andwould recommend it for the clinical environment"Eamonn O'Driscoll, Biochemical Genetics UnitCAMBRIDGE UNIVERSITY HOSPITALS NHS FOUNDATION TRUSTAMINO ACID ANALYSIS IN CLINICAL AND PHYSIOLOGICAL SAMPLESInvestigating metabolic disorders is a crucial and complex task.With over 42 clinically significant amino acids and their derivativesimplicated in the clinical diagnosis and monitoring of Inborn Errorsof Metabolism 1 , clinical laboratories need an accurate and reliableanalysis system that delivers unequivocal results quickly.The Biochrom 30+ is the latest gold standard dedicated amino acidanalyzer used by reference laboratories, hospitals, pharmaceuticaland industrial labs worldwide. It allows users to accurately detectand quantify amino acids and their derivatives in complexphysiological samples for both in vitro diagnostic use and research.FIVE GOOD REASONS TO BUYA BIOCHROM 30+ FOR CLINICAL ANALYSISQUALITY ANALYSISThe Biochrom 30+ is capable ofseparating up to 56 detectablephysiological amino acids, derivativesand other compounds present inbiological fluids including rare aminoacids such as alloisoleucine,sulfocysteine, and argininosuccinic acid.It gives clear separations with precision(cv.

amino acid analyzersA DEDICATED SYSTEMThe Biochrom 30+ is fully automatedwith an 84 position air cooledautosampler. It is designed to operate24/7 and ideal for busy clinical andresearch labs. This compact instrumentcan handle plasma, serum, urine, driedblood spots and CSF with low matrixinterference and without costly andtime-consuming re-equilibration.COST EFFECTIVEThe Biochrom 30+ is a cost effectivesolution for amino acid analysis becauseit has a long column life-designed to lastthe life of the instrument. With ourunique column cleaning and re-packingservice you can reduce costs further.There is a full range of competitivelypriced ready to use reagents that savespreparation time and minimizesvariation. All reagents are stable at roomtemperature simplifying storage.EASY TO USEAnalysis times can be tailored to thelaboratory’s requirement, with ready touse short methods. The Biochrom 30+has pre-defined analytical, processingand reporting methods in the softwareas well as dedicated technical support,training and installation to get your labup and running quickly.PEACE OF MINDThe Biochrom 30+ is backed by adedicated technical and engineeringsupport team. Our applications teamcan help with full screening methods orspecific short methods from ourapplication database built over manyyears. Our service contract package givescomplete peace of mind and includes:• Two maintenance visits per yearperformed by trained and certifiedBiochrom field service engineers.• Biochrom quality parts used for allmaintenance and repairs.• Rapid service from our engineeringteam.For more than 40 years, Biochrom hasbeen supporting consultants,pediatricians and clinical expertsthroughout the world by providing acomplete package for amino acidanalysis, from instrumentation andchemical kits, to customer trainingcourses and user group meetings.3

THE BIOCHROM 30+ CLINICAL AND PHYSIOLOGICAL SYSTEM4A CHROMATOGRAPHIC SYSTEMSPECIFICALLY FOR AMINO ACIDSThe Biochrom 30+ Amino AcidAnalyzer is a cation exchangechromatography system coupled witha highly specific detection systemusing post column derivatization withninhydrin reagent. Amino acids areseparated according to their netcharge determined by the pKa of theirionized groups. The mobile phase is afinely tuned set of 5 lithium citratebuffers used in a stepwise elutionprofile of increasing pH and molarity. Atemperature gradient on the columnmaximizes resolution. Lithiumhydroxide solution regenerates theresin bed online after each run cycle.HIGHLY SPECIFIC DETECTION SYSTEMThe ninhydrin method is highly specificbecause it reacts only with aminogroups giving a compound absorbingat 570nm wavelength (440nm foramino acids like proline). This responseis a linear relationship between theabsorbance and the amount of aminoacid in the sample. The sensitivity ofthe ninhydrin reaction is optimized forclinical use and the response is 100%linear within the expected amino acidsconcentration range encountered inclinical samples. The only samplepreparation needed is a simpledeproteinization-filtration step. Thecontinuous flow of reagent ensures areproducible derivatization giving highprecision in the peak area.THE BIOCHROM 30+AMINO ACID ANALYSIS SYSTEM• Biochrom 30+ Analyzer with aircooled autosampler• Choice of column(with top-up resin)• Starter pack of ready to use reagents• Spare parts and consumables kit• HP computer and monitor,HP printer, Windows ® operatingsystem and all cables• Biosys software and Ezchrom Elitedata handling software• Manuals and qualification &performance verification logbook• On-site customer trainingROBUST AND STABLE CHEMISTRYThe chemical kit contains everythingyou need for routine analysis of up to260 runs (Accelerated method).Chemicals are available either ascomplete kits or as individualbuffers and consumables toenable continuity of analysis.All reagents are stable atroom temperature andguaranteed to giveaccurate and reproducibleresults with a 3 year shelflife. On the instrument,buffers and reagents arestored under an inert gas to ensurestability.CHOICE OF RE-USABLE COLUMNS• High Speed Column - for rapid fullscreening• High Performance Column - thestandard reference• High Resolution Column - formaximum separationManufactured from PEEK material, thecolumns are free from corrosion andmetal contamination and are packedwith optimally sized cation exchangeresin. The columns are attached withfinger-tight fittings so no specialspanners are required to ensure a leakfreeseal. All our columns are fullytested and optimized under strict QCcriteria. To minimize waste and reducecosts, our columns are fully recyclable atthe end of their life thanks to ourunique repacking and cleaning service.

amino acid analyzersSOFTWAREBiochrom BioSys V. 3.0 softwarecontrols the system via an advancedgraphical user interface showing realtime operational information and is fullyintegrated with the data-handlingsoftware EZChrom Elite from Agilent.This powerful, advanced software iseasily networked for secure datastorage and includes special tools forcompliance with regulatory demands,e.g. FDA 21CFR11 part 11. It offers awide variety of functions to set upmethods and sequences, process andvisualize data and compile customreports. Data can be exported easily toother programs or other Windows ®applications. Other chromatographysystems use this software so itsfamiliarity may increase efficiency andhelp to reduce user-training costs.QUALITYThe Biochrom 30+ System is certifiedfor in vitro diagnostic use for thediagnosis of PKU. Biochrom reagentsare manufactured from high qualitymaterials under the ISO 9001:2008Quality System and subjected torigorous control procedures. Inaccordance with the IVD MedicalDevices directive 98/79/EC and the UKMedical Devices Regulations 2002 No.618. FDA exempt contact Biochrom forinformation.5

“Our Biochemical Genetics laboratory had the privilege ofpurchasing the first Biochrom Amino Acid Analyzer in theUnited States. We have been a Biochrom customer for 10years and have since purchased a second system. We havebeen very impressed with the outstanding service Biochromhas continually provided. The analyzers are reliable, rugged,and consistently produce high quality data that enables us toreport accurate results for inborn metabolic diseases.”CLINICAL ANALYSISErin T. Strovel, Ph.D., F.A.C.M.G.Director Pediatric Biochemical Genetics LaboratoryAssistant Professor of PediatricsUNIVERSITY OF MARYLANDThe Biochrom 30+ enables you to recognize a characteristic amino acid chromatographic profile to aid diagnosis. The quantificationof the amino acids may indicate the severity of the abnormality detected. For reference purposes, chromatograms of normal plasmaand urine samples are shown.6Amino acid standard analyzed on the Biochrom 30+ accelerated method.

PLASMA ANALYSISA full screening is achievable in less than 2 hours (injection to injection) in physiological samples with the Accelerated System.Rare key markers are clearly separated as shown in the plasma sample below.mVoltsPlasma sample spiked with saccharopine, alloisoleucine and argininosuccinic acid.URINE ANALYSISUrine sample analyzed on the Biochrom 30+ High Performance System.7

PHENYLKETONURIA (PKU)The PKU method is designed specifically for PKU newborn screening and to monitor patients’ phenylalanine levels in plasma.Samples can be analyzed within 30 minutes with an injection-to-injection repeat time of 50 min. The method is compatible withdried blood spot samples and gives cost effective PKU screening service.Normal plasma sample analyzed on a Biochrom 30+ equipped with the PKU column. Norleucine used as theinternal standard.MAPLE SYRUP URINE DISEASE (MSUD)Maple Syrup Urine Disease (MSUD) can be diagnosed by the presence of alloisoleucine combined with high amounts of leucine,isoleucine and valine 2 . The Biochrom 30+ separates fully isoleucine, leucine and alloisoleucine enabling a clear diagnosis.8MSUD patient sample analyzed with the MSUD short method. Data kindly provided by Tanyalcin Laboratory, Turkey.

TOTAL HOMOCYSTEINE ASSAYElevated plasma total homocysteine has emerged as an important risk factor in the development of vascular disease 3 . Using theBiochrom 30+ Amino Acid Analyzer reduces analysis time to 24 min injection to injection for evaluation of patients at risk.Physiological standard spiked with homocysteine. Norvaline is used as the internal standard.SULFOCYSTEINESulfite oxidase deficiency is a rare inborn error of metabolism, characterized by elevated urinary and plasma s-sulfocysteine 2 . Usingthe Biochrom 30+ Amino Acid Analyzer, a short programme for the analysis of s-sulfocysteine has been developed that allow itsseparation from phosphoserine.mVoltsPatient sample urine sulfocysteine. Data kindly provided by St James Hospital, Leeds UK.9

Biochrom 30+ TECHNICAL SPECIFICATIONSReproducibilityLimit of detectionArea: Better than 0.5% RSD at 10 nanomoles.Retention time: Better than 0.1%RSD9 - 15 pmoles NinhydrinTime of analysis (based on the separationof 45 physiological amino acids):Accelerated analysis:High Performance analysis:115 min injection to injection170 min injection to injectionAnalytical column:High pressure PEEK column packed with Ultropac 8 cation exchange resin. Peltierheating/cooling system.Eluent system:6 buffer system (5+1 regeneration solution) stored on the instrument at roomtemperature in graduated 1L glass bottles under nitrogen pressure.Ninhydrin reagent: Stored on the instrument at room temperature undernitrogen pressure in a 2L plastic coated glass bottle.TemperaturePhotometric detection:Sample injectionColumn temperature variable between 20°C and 99°C. Reaction Coiltemperature adjustable between 40°C and 145°C (135°C is optimum).Single flow cell with optical beam splitter.Dual channel detection at 440nm and 570nm.3 injection modes (full loop, partial loop and micro), 84 position autosampler.Sample volumes from 1µL to 5000µL. 200µL loop supplied as standard.Software:BioSys v3.0 control softwareBiochrom Alias Manager autosampler control softwareLatest version of EZChrom Elite Data Handling 21 CFR part 11 compliantDimensions and weights:Bench top Fluidics Cabinet: 48 x 59 x 57 cm, 19 x 23 x 22 inches (w x d x h)Weight:50kg, 110lbsAutosampler: 30 x 57.5 x 36 cm, 12 x 23 x 14 inches (w x d x h)Weight:21kg, 46lbsOperating Conditions:Operating temperature: 15 °C to 25 °CMaximum humidity: 80% at 25 °CRequired services:Safety SystemOxygen free nitrogen gas (99.99%) or Argon regulated to 73.5psi (5bar).Drainage facility. 240V/100V, 50Hz/60Hz, 300VA mains supply.Automatic shut-down and reaction coil flushing in the event of:n photometer lamp failuren incorrect ninhydrin / buffer / coil / nitrogen pressuresn incorrect coil and column temperaturesn power failureOrder CodeBiochrom 30+ Physiological System80 - 6000 - 50

ELUTION ORDER OF CLINICALLY SIGNIFICANTAMINO ACIDS AND DERIVATIVES ANALYZEDBY THE Biochrom 30+1 Sulfocysteine2 Phosphoserine3 Taurine4 Phosphoethanolamine5 Urea6 Aspartic acid7 Hydroxyproline8 Methionine sulfone9 Threonine10 Serine11 Asparagine12 Glutamic acid13 Glutamine14 Sarcosine15 Cysteine16 α-Aminoadipic acid17 Proline18 Glycine19 Alanine20 Citrulline21 α-Aminobutyric acid22 Valine23 Homocysteine24 Cystine25 Saccharopine26 Pipecolic acid27 Homocitrulline28 Methionine29 Cystathionine30 AlloIsoleucine31 Isoleucine32 Leucine33 Argininosuccinic acid34 NorLeucine35 Cysteine-homocysteine mixed disulfide36 Tyrosine37 β-alanine38 Phenylalanine39 δ-Aminolevulinic acid40 β-Aminoisobutyric acid41 Homocystine42 γ-Aminobutyric acid43 Ethanolamine44 Ammonia45 Hydroxylysine46 Amino Ethyl Cysteine47 Ornithine48 Lysine49 1-Methyl histidine50 Histidine51 Tryptophan52 3-Methyl histidine53 Anserine54 Carnosine55 Arginine56 HomoarginineABOUT BIOCHROMBiochrom is a world leader in amino acid analysis. The Biochrom 30+ is recognizedas the gold standard dedicated amino acid analyzer used by hospitals,pharmaceutical and industrial labs worldwide. Applications are available both forclinical analysis and screening of metabolic disorders, for drug synthesis, infusionfluids and for industrial applications in food, beverage and feedstuffs.Biochrom is a leading manufacturer of scientific instruments with over 40 year’sexperience. The Biochrom Group consists of five well-known instrument brandscovering amino acid analysis, UV/Vis spectroscopy, and microplate instrumentation.Hospitals and laboratories worldwide trust our products and we are a valued OEMpartner of many of the world's finest scientific instrumentation companies. TheBiochrom spectroscopy range includes the Novaspec, Ultrospec ® , and GeneQuant ® ,plus the Biochrom Libra and Biochrom WPA brands. Biochrom also manufacturestwo major brand names in the microplate instrumentation market - Biochrom Asysand Biochrom Anthos.All our instruments are available through a growing global network of independentdistributors, backed by our commitment to customer support. Biochrom is a HarvardBioscience Company.EZChrom Elite is a trademark of Agilent TechnologiesWindows is a trademark of MicrosoftHP is a trademark of Hewlett PackardUltrospec and Genequant are registered trademarks of Biochrom1Differential diagnosis of (inherited) amino acid metabolism or transport disorders.W. Blom & J.G.M.Huijmans, Amino Acids (1992) 2:25-672Vademecum Metabolicum Manual of Paediatrics. J. Zschocke & G. Hoffmann, 2ed.Milupa GmbH 20043Rapid Analysis of Homocysteine Levels. A.Lolia & S. Bee,Biochrom Application note B30.5.11

Biochrom Ltd22 Cambridge Science Park,Milton Rd, Cambridge CB4 0FJ UKTel: +44 (0)1223 423723Fax: +44 (0)1223 420164Email: enquiries@biochrom.co.ukwww.biochrom.co.ukDistributors worldwideBiochrom US84 October Hill Road,Holliston, MA 01746-1388 USATel: (Toll free): 877- BIO-CHROM (877-246-2476)Fax: 508-429-5732email: sales@biochrom-us.comwww.biochrom-US.com80-6000-42 Issue 1