ALGA AE AND D BIOFU UELS FACILI F ITY Hosted by Soutth Austrralian Re esearch and Develop pment In nstitute – Aquattic Scien nces This state of tthe art analytical and algal biomass produc ction facility y was suppported by fu unding from m the Nattional Colla aborative Research R In nfrastructurre Strategy y (NCRIS) and a the Edducation Inn novation Fu unds (EIF) under th he Super Science S Initiative and h hosted by the t South Australian A Research and Dev velopment Institute (S SARDI) at their t Aquattic Science es division at a West Beeach, SA. The T service es pro ovided by th his facility are a available to resea archers from m public se ector instituutions, universities and d industry on a fee for servvice basis. ANA ALYTIC CAL CA APABILITY PROTEIN P A ANALYSIS S Extraction E n • • • • Chemiical extracttion of Tota al Proteinss using Triz zol Preferrred form of samples - homogen neous dried powder Minimu um sample e size = 0.5 5g Minimu um numbe er of samplles = 10 Quantifica Q ation • Quantiification of Total proteein contennt • Coomaassie Braddford spectrophotom etric assayy • Minimuum numbeer of samplles = 10 Character C risation • Agileent 2100 bbioanalyser • Sizinng and quaantitation of o proteins based on on-chip ge el elec ctrophoresiis • Miniimum num mber of sam mples = 10 FATTY F AC CID PROFIILES Quantifica Q ation by Gas Chrom matograph hy – Flame e Ionizatio on Detecto or (GC-FID D) and Identificati I ion by Ga as Chroma atographyy – Mass Spectrome S etry (GC-M MS) • Perkkin Elmer G GC-MS (Clarus 560s s) for Fatty acid methylated este er • • • • • (FAM ME) identiffication Perk kin Elmer G GC-FID (C Clarus 500) for quantiffication Exte ernal stand dards conta aining 30 FAMEs F useed for qualitative anallysis Prefferred form m of sample es - homog geneous drried powde er Miniimum samp ple size = 0.5g Miniimum num mber of sam mples = 1 TOTAL T LIP PID CONT TENT Hot H Soxhllet extractiion and qu uantificatiion • Gerhardt Soxttherm, autoomatic hot soxhlet syystem for tootal lipid dete ermination in solid sa amples • Prefferred form m of sample es - homog geneous drried powde er • Miniimum sampple size = >5g > • Miniimum num mber of sam mples = 6 Gravimetr G ric low tem mperature extraction n and qua antification n • Buchi Rotavappor R210, Buchi V7 700 and V V850, prov vides a miild apprroach to tthe determ mination of o total lippids in so olid sample es without excesssive heatin ng. • Prefferred form m of sample es - homog geneous drried powde er • Miniimum sampple size = 5g • Miniimum num mber of sam mples = 1 Modified M B Bligh & Dyyer extraction and g gravimetric quantific cation • • • • Modifie ed Bligh & Dyer technique for to otal lipid estimation Ideal fo or sample size < 5g and a or whe ere lipid co ontent is prredicted to be < 5% Estima ates crude total lipid content c Minimu um numbe er of samplles = 6 LIPID L CLA ASSES Qualitative Q e and qua antitative analysis a o of lipid clas sses • • • • • Includess Triglycerides, Phos spholipids a and Free Fatty F Acids s Qualitattive analysis using GC Quantita ative analyysis using Single S Pha ase Extracttion (SPE) columns Preferre ed form of samples s – extracted samples or o homogeneous drieed powder Minimum m number of samples = 4 TOTAL T CA ARBOHYD DRATES Total T carbohydrate quantificat q tion • • • • • Pheno ol-sulfuric acid a spectro ophotomettric assay for f Total ca arbohydrattes Calibra ated againsst standard d glucose Preferrred form off samples - homogen neous dried powder Minimu um sample e size = 0.5 5g Minimu um numbe er of samplles = 6 GLYCOGE G EN Glycogen G quantifica ation • • • • • Spectrrophotome etric quantiffication usiing Anthrone techniq que Calibra ated against standard d glycogen n Preferrred form of samples - homogen neous dried powder Minimu um sample e size = 1g g Minimu um numbe er of samplles = 12 Page 2 of 8 CALORIFI C C VALUE (ENERGY Y ESTIMAT TION) Calorimete C er Energyy • IKA Oxygen boomb caloriimeter calibrated agaainst Benzooic acid • • • • stan ndards Dete ermines th e gross ca alorific valu ue for liquidd and solid samples Prefferred form m of sample es - homog geneous drried powde er Miniimum samp mple size = 0.6g Miniimum num mber of sam mples = 6 TOTAL T CA ARBON AN ND NITRO OGEN ANA ALYSIS Total T orga anic carbo on, total inorganic ca arbon, tottal carbon, total bou und nitrogen • Analytic Jennaa Multi N/C C analyser • Analysis of TC C, TOC and d TIC in liq quid and soolid sample es and the TNb in liquid sa amples • Miniimum sampple size = 1g 1 carbon only o / 10mll liquids • Miniimum numbber of sam mples = 6 Total T Kjeld dahl Nitrog gen and Protein P • • • • • Tota al Kjeldhal Nitrogen in solid and d liquid sam mples Buchi Total Kje eldahl Nitrrogen Analyser Crud de protein (calculated d) from TK KN values Miniimum samp mple size = 1g Miniimum numb ber of sam mples = 4 ELEMENT E TAL ANALY YSIS Inductively I y Coupled d Plasma – Optical Emission Spectrom metry (ICP P-OES) • • • • • ICP--OES Varia an Vista-P PRO Trac ce and maccro-elemen ntal analys sis in liquidd and solid samples Solid d samples will requirre digestion n. Miniimum samp ple size = 7ml of liqu uid / 1g of ffine powde er Miniimum num mber of sam mples = 6 INORGANIC DISSOL LVED NUT TRIENT AN NALYSIS Ammonium A m, Nitrate e, Nitrite, Total T Oxid dised Nitro ogen, Urea a, Phosph hate, Silica ate • Aquakem 200ccd discretee nutrient analyser a • Deteermination of the nutrient conce entrations in clean water (fresh h, • • • • sea or brackis h), wastew water, proc cess water,, effluents and growth h med dia. High h throughp put ~60 sam mples per analate a peer hour Inclu udes Nitratte, Nitrite, Total Oxidised Nitroggen, Urea, Phosphatte, Amm monia, Silicca Miniimum samp mple size = 50ml Miniimum numb ber of sam mples = 10 Page 3 of 8 RADIOISO R OTOPE QU UANTIFICA ATION 14 1 C, 3H, 32P and 33P analysis • Perkkin Elmer TTri Carb 29900tr liquid d scintillatioon counterr • Deteection and counting of o alpha, beta and gaamma radioactivity • Proccessed sam mples in liq quid scintillation cockktail in scin ntillation via als TITRIMETR T RIC ANAL LYSIS Total T alkalinity • • • • • Me etrohm Ele ctrotitratorr (719 Titrin no) Endpoint titra ation with a strong ac cid Total alkalinitty correspo onds to titrable alkali nity as pH 4.5 Min nimum sam mple size = 100ml filtered Min nimum num mber of sa amples = 10 0 Acid A value e and free e fatty acid d percenta age (%FFA A) of biodiesel, fatss and oils • • • • Qu uantification n of acidity y and %FFA of biodieesel, fats and a oils Co olourimetricc titration with w a stron ng base Ep ppindorf Mu ultipipette X-Stream X Min nimum sam mple size = 30g (acid value annd %FFA), 3g (%FFA A only) • Minnimum num mber of sa amples = 1 Karl K Fishe er moisture e determin nation • 831 KF Couloometer, for the Karl Fisher F deteermination of moisturre con ntent in bio odiesel, fatts and oils • Minnimum sam mple size = 10g • Minnimum num mber of sa amples = 5 MOISTURE M E / ASH / TOTAL T SU USPENDE D SOLIDS S / PARTIC CULATE O ORGANIC MATTER / PARTICUL P LATE INOR RGANIC MATTER M A ANALYSIS S Moisture M A Analyser • Metttler Toledoo HG63 mooisture ana alyser • Rappid and preecise methood to deterrmine moissture conteent in solid sam mples • Miniimum samp mple size = 1g • Miniimum num mber of sam mples = 1 Gravimetr G ric determination of ash • • • • • Gra avimetric d determination of mois sture/ash ccontent Co ombustion in a muffle e furnace at 500-550°°C Further analyysis for acid soluble and a acid innsoluble an nalysis Min nimum sam mple size = 1g Min nimum num mber of sa amples = 10 0 Page 4 of 8 ROTENON R NE ANALY YSIS High H Perfo ormance Liquid L Chrromatogra aphy • Shim madzu HP PLC system m equipped with Ph otodiode Array A (PDA A) • • • • and refrigerate ed autosam mpler Qua antification of rotenon ne in solid and a liquid samples Calibrated aga ainst roteno one standa ard Miniimum samp mple size = 1g Miniimum num mber of sam mples = 1 ADD DITION NAL EQ QUIPME ENTS • Flow cyytometer (B Beckman-c coulter Ce ll lab Quan nta SC) • 18L Freeeze Dryer (Labconcco) • Milli Q Integral Water W Purific cation Sysstem (TOC <5ppb) an nd a Millipoore Elix 5 Reverse R Osmossis system • • • • • 86L -80 0ºC Freeze er Refrige erated centtrifuge (Eppendorf 58 810 R, max ximum cap pacity 4 x 2250ml) Muffle furnace (1100ºC) 2 x fum mehoods Varietyy of precisio on balance es from 0.0 01mg – 4 kg k resolutio on PHOTOSY P YNTHETIC PIGMENT TS High H Perfo ormance Liquid L Chrromatogra aphy – Pig gment pro ofile suite • Shim madzu HPL LC system m equipped with Fluorrescence detector d an nd refrigerated au utosamplerr • Photosyntheticc pigment profiles fro om solid orr liquid sam mples • Calibrated usinng 19 phottosyntheticc pigment sstandards • Miniimum samp mple size = 0.2g Spectroph S hotometer/ r/ Spectroffluoromete er / Microp plate Rea ader - Qua antification of pigments • Synergy 4 Miccroplate reaader, Shim madzu UV-11700 • • • • Spectrophotom meter and Hitachi F2500 Specttroflurometter Qua antification of Chlorop phyll a, b, c and total carotenoid ds Acetone / Etha anol extrac cted samples Miniimum samp mple size = 500mg of dried powdder or sam mple supplie ed on 0.2 0 µm pore e size cellulose aceta ate or glasss fiber filterr paper Miniimum num mber of sam mples = 3 Page 5 of 8 PHOTOPH P HYSIOLOG GICAL ASS SAYS Walz W Phytto-PAM an nalyser • Phyto-PAM m measures emitted e fluorescencee at 4 excitation wa avelengths • Assessment of photosyynthetic performancee and light adaptationnal sta ate of phyto oplankton • Samples mayy be microoalgal cultures or natuural phytopplankton sam mples Walz W Divin ng PAM analyser a • Unnderwater ffluorometeer • In situ s measu urements of o chloroph hyll fluoresccence • For use on seeagrassess, macroalg gae, coral zzooxanthelle Aquation A S Submersible Photophysiolog gical Syste em • Divver deployeed submerrsible cham mber systeem with shu utter • • • • • fluo orometers and senso ors for PAR R, temperaature, pH and a salinityy Da ataloggers and powerr modules allow for uunattended d operation ns Pumps for m ixing and flushing f In situ s measu urements photosynth p hesis For use on se eagrasses s and macrroalgae Set of 10 cha amber systtem includiing 2 blankk chambers WATER W QUALITY ANALYSIS A pH, p conductivity, temperatu t ure, carbo on dioxid de (CO2), dissolveed oxygen n (DO), light, density, d viiscosity nt of param meters inclu uding: • Meeters availaable for meeasuremen pH, conductivity, temperature, CO2 , DO, lightt, density, visco osity Page 6 of 8 ALGAL PRODUCTION SY YSTEMS S SMALL S LA ABORATO ORY SCAL LE EXPERIIMENTAL SYSTEM • 12 x Te eco Aquarrium heater/chillers (8 800L-2000 0L capacity y) • Aquatrronica conttrol and mo onitoring ssystem with h pH and te emperaturee sensors • 24 x 20 00W LED lights – Pla ant lights ccapable of yielding up p to 1500μ μmol.m-2.s-1 PAR MASS M CUL LTURE INFRASTRU UCTURE Pure P culture and iso olation faciility Inclu udes: • Laminar Fllow Cabine et • Petriplate ccolony Cou unter • Fluorescen nt microsco ope (Olympus BX-611) and Digital inverted d microscop e (EVOS) for digital imaging, qquantificatio on of intracellula ar lipid prod duction and automateed cell cou unts • 110L Autocclave (Hira ayama HV) Controlled C d Environm ment Room m • Co ontrolled en nvironment room with temperatuure and ligh ht control and CO O2 input • 2 x light and ttemperaturre controlle ed growth ccabinets Micro M race eway syste em (250L, 2.5m2) 2 • 2 x 2.5m 2 race eway ponds s including control sysstems for CO C 2 injectio on and pH contro ol R&D R scale e racewayy system (3 3500L, 20 0m2) • 9 x 20m 2 2 racew way ponds including automated a control and mon nitoring sysstem for CO O2 injection, pH, waterr level, salinity and DO O mon nitoring for cultivation of micro- and a macroaalgae Bench B top p bioreacto or (12L) • 12L bench top Applicon fermentor f in ncludes coontrol syste ems for temp perature, p pH, DO, CO O2 injection, nutrient feeed, data lo ogging and d mixing. Bolt on n lights ava ailable for use as a phhotobioreac ctor. Ideal fo or stud dies on auto otrophic, mixotrophic m and heterootrophic systems. Large L scalle - closed d system (3500L) • 3 x Pilot-Scale photobio oreactor comprising off a 3.5m3 Algaelink A sollutions tubu ular reactor, with auto omated reggular injection of nuttrients and CO2 Page 7 of 8 DOWN D STREAM PR ROCESSIN NG EQUIPM MENTS • Bio o-ULTRA iss a pilot sca ale automa ated biodiessel reactorr that uses ultrrasound to enhance the t transes sterificationn of methan nol and oil to t form high qua ality biodies sel under controlled c teemperature e. • 2 x Clara 80 IIndustrial process p cen ntrifuges • Us sed for harvvesting alga al biomass s from masss culture sy ystems • Associated in nfrastructurre includes, seawaterr intake, filtrration and dis sinfection, ssedimentattion tanks and a storagee for algal biomass b • My yers vacuum m centrifug gal molecullar distillatioon system for sep paration an nd purificattion feed materials succh as oils • Ca an be applie ed for puriffication of Polyunsatu P rated Fatty y Acids from m alg gal lipids • Als so used in cchemical, food f proces ssing, pharrmaceutica al and pettroleum pro ocessing Con ntacts: Drr Sasi Nay yar Fa acility Mana ager Te el: +61 8 82 207 5321 Em mail: sasi.n [email protected] gov.au Mrrs. Michellee Braley Laboratory M Manager Tel: +61 8 82207 5330 E mail: m micheelle.braley@ @sa.gov.au u Page 8 of 8 ALG GAE AND BIOFUELS FAC CILITY SARDI Aquatic Scie ences 2 Hamra A Avenue West Beach SA A 5024 P: (08) 88207 5321 F: F (08) 8207 7 5481
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