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1. BioPak point of use ultrafilter Final purification step at the ultrapure water point of delivery provides pyrogen and nuclease free water at a high flow rate when you need It The BioPak unit is a disposable ultrafiltration cartridge typically used in cell culture biochemistry or molecular biology applications It can be installed at the outlet of Type water purification systems such Key benefits e Direct connection to all EMD Millipore Type water systems as the Milli Q Direct Q Simplicity or Synergy Pyrogen free water lt 0 001 EU mL production systems to produce pyrogen and nuclease free e RNase free water lt 0 01 ng mL and ultrapure water for a period of up to three months DNase free water lt 4 pg uL production The cartridge is composed of polysulfone hollow fibers e Safe method that eliminates the need to in a white ABS housing The BioPak ultrafiltration treat water with DEPC membrane is designed to optimize the rejection of e Bacteria free water lt 1 cfu mL production e Warranty of results within specifications for a minimum of 90 days usage e Maintenance free pyrogens nucleases and bacteria while maintaining a high flow rate and minimizing the release of ionic and organic materials EMD Millipore is a division of Merck KGaA Darmstadt Germany EMD MILLIPORE PYROGEN REMOVAL The most common pyrogens are endotoxins i e lipopolysaccharides LPS from the walls of2. Gramnegative bacteria The LPS have two major parts a hydrophilic polysaccharide chain with antigenic regions and a hydrophobic lipid group As the polysaccharide chain is variable in length the LPS molecular weight ranges from 3 000 to 25 000 Dalton In ultrapure water the LPS sub units aggregate to form higher molecular weight structures that can be removed by ultrafiltration membranes with cut off below 20 000 Dalton Pyrogens are known to affect cell culture and biochemistry experiments in numerous ways It has been demonstrated that their interaction with cell membranes causes morphological changes and damage as well as the secretion of specific substances such as tumor necrosis factor cytokines or enzymes Pyrogens also affect the cell division process enhancing or reducing it depending on the nature of the cell line The presence of pyrogens also may affect analytical techniques such as electrophoresis For these reasons it is good laboratory practice to remove pyrogens from all solutions used in cell culture and other biochemical applications Experiments performed in EMD Millipore R amp D laboratories have demonstrated that the BioPak ultrafiltration cartridge can be used for at least 90 days to treat Milli O ultrapure water and obtain product water with a pyrogen level below 0 001 EU mL NUCLEASE REMOVAL Challenge tests performed in EMD Millipore R amp D laboratories have demonstrated that the BioPak cartridge all
3. connectors for connection to Millipak final filter For more information please visit our website www millipore com laowater Millipore Milli Q Direct O Synergy Simplicity and Millipak are registered trademarks of Merck KGaA Darmstadt Germany VOC Pak the M mark and EMD Millipore are trademarks of Merck KGaA OI Corporation is a trademark of OI Corporation Agilent Technologies is a registered trademark of Agilent Technologies Inc Lit No PB1632ENUS 01 12 2012 EMD Millipore Billerica MA U S A All rights reserved
4. lt 0 05 isopropylbenzene lt 0 05 4 isopropyltoluene lt 0 05 pe atl lt 0 10 naphtalene lt 0 10 n propylbenzene lt 0 50 styrene lt 0 10 1 1 1 2 tetrachloroethane lt 0 05 1 1 2 2 tetrachloroethane lt 0 10 tetrachloroethene lt 0 05 toluene lt 0 10 1 2 3 trichlorobenzene lt 0 05 1 2 4 trichlorobenzene lt 0 05 1 1 1 trichloroethane lt 0 10 1 1 2 trichloroethane lt 0 05 trichloroethene lt 0 05 trichloromonofluoromethane lt 0 10 1 2 3 trichloropropane lt 0 05 1 2 4 trimethyloenzene lt 0 05 1 3 5 trimethyloenzene lt 0 10 o xylene lt 0 05 m xylene lt 0 05 p xylene lt 0 05 vinyl chloride lt 0 20 OL Quantification Limit expressed in ppb ug l CCI C H CI 4 8 Molecules removed by the VOC Pak cartridge include carbon tetrachloride CCI and chlorobenzene C H CI g Milli O water purification system fitted with a VOC Pak cartridge CHCI C H CI P Molecules removed by the VOC Pak cartridge include chloroform CHCI and vinyl chloride CHCI Direct Q 3 water purification system with VOC Pak cartridge Ordering information Description Catalogue No VOC Pak cartridge 1 pk delivered hermetically sealed with a Certificate of Quality VOCPAKOO1 Cartridge installation and Conditioning Kit including the following reusable parts Polyethylene 1 4 Gaz F Hose barb connection with O ring EDSKITOO1 e Polyethylene 1 4 Gaz F 1 4 Gaz F
5. lt x 1 2x108 140000 0 016 6 0x10 70000 0 000 0 0 0 0 00 8 00 16 00 24 00 32 00 40 00 5 00 10 00 15 00 20 00 25 00 30 00 35 00 40 00 0 00 8 00 16 00 24 00 32 00 40 0C Minutes Minutes Minutes Bottled water F for LC MS Bottled water J for LC MS Bottled water B for LC MS Milli Q Integral with LC Pak EQUIPMENT The equipment used throughout the experiment was from Waters Corporation Milford MA U S A e LC system Alliance 2695 e Pre column Column C18 X Terra 3 um 4 6 x 30 mm e Column C18 Atlantis 3 um 2 1 x 150 mm e Detectors PDA Model 2996 e Mass detector ZQ 2000 e Elution solvents Water to be tested Acetonitrile HPLC grade J T Baker e Computer for data acquisition and system control equipped with Empower software Detailed analytical conditions available upon request internal procedure No 00084229S0 The entire study including details of analytical procedures is provided in the LC Pak cartridge Validation Guide ORDERING INFORMATION Description Comments LC Pak Cartridge 1 pk delivered hermetically sealed with a Certificate of Quality pore Cartridge installation and Conditioning Kit including the following reusable parts e Polyethylene 1 4 Gaz F Hose barb connection with O ring EDSKITOO1 e Polyethylene 1 4 G F 1 4 Gaz F connectors for connection to Millipak Millipak 0 22 um end filter MPGP04001 Millipore Milli Q Direct Q Synergy LC Pak Simplicity and Mil
6. C MS e Each LC Pak cartridge is delivered with a Certificate of Quality Recent advances in analytical techniques such as UPLC LC MS LC MS MS have considerably improved the sensitivity of organic and biochemical molecule detection As a result these techniques now require improved water quality for the production of mobile phase buffers blanks standards preparation sample dilution glassware rinsing or extraction EMD Millipore is a division of Merck KGaA Darmstadt Germany EMD MILLIPORE Although fresh Milli O water has proved adequate for most procedures involving these techniques until recently there is now a growing need for higher water quality in some advanced applications To answer this emerging need EMD Millipore has developed a new POD Pak designed to produce ultrapure water with an organics contamination a level below most water sources available today at a fraction of the cost The LC Pak cartridge can be connected to the outlet of existing and new EMD Millipore Type water systems Milli Q Direct O Synergy and Simplicity to produce water with a TOC level below 5 ppb and deliver Ea at least 500 L of ultrapure water with minimum trace a j organic contamination The LC Pak cartridge uses the pia well known reversed phase silica purification media to remove traces of neutral organics A LC PAK CARTRIDGE DESIGN The LC Pak is designed to provide scientists with the cer
7. LISHER SPECIFICATIONS The VOC Pak cartridge is designed to producewater in which the VOC concentration is below the analytical quantification limit for the VOCs listed in Table 1 provided that the VOC concentration in feed water does not exceed 1 ppb 1 ug l and the Total Oxidizable Carbon TOC of the ultrapure water entering the VOC Pak cartridge is below 5 ppb In order to warrant this water quality the VOC Pak cartridge was qualified by a challenge test the VOC Pak cartridge was fed by ultrapure water containing a 1 ppb 1 ug l concentration of the VOC to be eliminated The concentration of the VOC was measured by Gas Chromatography GC downstream of the VOC Pak cartridge after passage of 50 100 150 200 250 300 and 350 in order to determine the breakthrough point of the different VOCs The results are displayed in Table 1 for a processed volume of 350 I eE i amp k i ate Pm Q POD dispenser fitted with a VOC Pak cartridge Analysis of the VOCs in ultrapure water was performed according to EMD Millipore procedure 84214S0 using an Eclipse 4660 Purge and Irap sample concentrator from Ol Corporation College Station Texas and a GC MS system GC model 6890N and MS model 5975B from Agilent Technologies Inc Santa Clara California A copy of the complete qualification report with description of the analytical procedure is available upon request Table 2 Quantificat
8. ak unit has been individually tested for efficiency and flow rate and is delivered with a Certificate of Quality ORDERING INFORMATION Description Catalogue No BioPak Ultrafiltration Cartridge 1 pk validated for pyrogen and nucleasefree water production delivered with a self adhesive label with space to note CDUFBIOO1 installation and replacement dates Certificate of Quality and multilingual User manual Lit No PB1O06ENUS 01 12 Copyright 2012 EMD Millipore Billerica MA U S A Millipore Milli Q Direct Q Simplicity BioPak and Synergy are registered trademarks of Merck KGaA Darmstadt Germany The M Mark and EMD Millipore are trademarks of Merck KGaA All rights reserved Design Sysaxe LC Pak Point of use Polisher C18 Reverse phase silica Key Benefits e The LC Pak cartridge is easily connected to the outlet of all EMD Millipore Type water systems Mil li O Direct O Synergy and Simplicity e Final purification step uses efficient C18 reverse phase silica technology to provide fresh ultrapure water with low traces of organics at high flow rate when you need it e Delivers a minimum of 500 L of organic free ultra pure water e Designed to answer the demands of UPLC LC MS and LC MS MS analytical techniques used for organic trace and ultra trace analysis e Validated for the production of water whose quality equals or exceeds the specifications of bottled water for L
9. ay be only somewhat dangerous for adults mounting evidence Suggests that they can significantly disturb fetal and infant development As researchers increasingly focus on the effects of these chemicals there is a need for endocrine disrupter free water for use in experimental tests While conventional water purification techniques remove the bulk of organic compounds from ultrapure water some trace organics still may remain Examples of endocrine disrupters include nonylphenol a detergent bisphenol A a plasticizer diethylstiloestrol a growth agent administered to livestock and DDT a pesticide Examples of man made chemicals with estrogeno mimetic structure Estradiol AC These man made chemicals can be found in many products widely used in industrial societies e Bisphenol A is a plasticizer used in reusable water bottles laptop computer housings dental sealants and the resins in some food can linings e Phthalates are found in a wide variety of products including vinyl flooring food packaging blood storage bags intravenous medical tubing as well as many health and beauty products such as detergents soaps shampoos deodorants fragrances hair spray and nail polish They are often used as a plasticizer in PVC pipes distributing water in buildings AA Nonylphenol OH Q Diethylstilbestrol Bisphenol A Al CO EDS Pak polisher design The EDS Pak is a disposable point o
10. carbon tetrachloride 1 06 lt 0 05 lt 0 05 chlorobenzene 0 94 lt 0 05 lt 0 05 chloroform 1 07 lt 0 05 lt 0 05 dibromochloromethane 1 01 lt 0 10 lt 0 10 1 2 dichlorobenzene 1 01 lt 0 05 lt 0 05 1 4 dichlorobenzene 1 53 lt 0 05 lt 0 05 1 2 dichloroethane 1 08 lt 0 05 lt 0 05 1 1 dichloroethene 0 87 lt 0 05 lt 0 05 cis 1 2 dichloroethene 0 95 lt 0 05 lt 0 05 trans 1 2 dichloroethene 1 53 lt 0 05 lt 0 05 1 2 dichloropropane 0 86 lt 0 05 lt 0 05 cis 1 3 dichloropropene 0 97 lt 0 04 lt 0 04 trans 1 3 dichloro T PE r propene ethylbenzene 1 09 lt 0 05 lt 0 05 tetrachloroethene 1 44 lt 0 05 lt 0 05 toluene 0 96 lt 0 10 lt 0 10 1 1 1 trichloroethane 0 94 lt 0 10 lt 0 10 1 1 2 trichloroethane 0 93 lt 0 05 lt 0 05 trichloroethene 1 02 lt 0 05 lt 0 05 o xylene 0 79 lt 0 05 lt 0 05 m xylene 1 02 lt 0 05 lt 0 05 p xylene 0 94 lt 0 05 lt 0 05 Challenge VOC concentration in feed water upstream of VOC Pak cartridge Quantification limit limit of quantification of the VOC measured by GC Concentration of VOC below the VOC Pak cartridge for a feed water concentration of 1 ppb VOC Pak POLISHER OPERATION The VOC Pak cartridge is easy to use simply connect the VOC Pak unit to the outlet of the EMD Millipore ultrapure water system and flush 5 of water You are now ready to deliver at least 300 of VOC free ultrapure water VOC Pak PO
11. docrine Disrupters on Developmental and Reproductive Functions Current Drug Targets Immune Endocrine amp Metabolic Disorders Volume 5 Number 1 p 1 10 2005 3 Bigsby R Chapin R E Daston G P Davis B J Gorski J Gray L E Howdeshell K L Zoeller R T and vom Saal F S Evaluating the Effects of Endocrine Disrupters on Endocrine Function during Development Environmental Health Perspectives 107 Suppl 4 p 613 618 1999 01 12 Copyright 2012 EMD Millipore Billerica MA U S A Milli Q EDS Pak Synergy Simplicity Direct Q and Millipak are registered trademarks of Merck KGaA Darmstadt Germany The M mark EMD Millipore are trademarks of Merck KGaA All rights reserved VOC Pak Point of Use Polisher Final purification step at the ultrapure water point of delivery efficiently removes Volatile Organic Compounds VOCs EMD Millipore is a division of Merck KGaA Darmstadt Germany VOCs are industrial contaminants that may cause health safety issues and therefore should be detected in the environment VOCs are routinely analyzed in water soil soil gas and air Specifications for VOCs in drinking water have been established by different regulatory agencies in many countries A table showing the Maximum Allowable Concentration MAC for some of the different VOCs monitored in the United States Europe Japan and China as well as by the World Health Organization WHO is available on t
12. effi cient removal of the following endocrine disrupters bisphenol A diethyl phthalate di n buty phthalate and nonylphenol EDS tested Feed ppb Product ppt Volume I Flow l min Bisphenol A Up to 4 lt DL 5 ppt 300 0 5 2 0 Diethyl phthalate Up to 1 5 lt DL 200 ppt 300 0 5 2 0 Di n buty phthalate Up to 1 5 lt DL 200 ppt 300 0 5 2 0 Nonylphenol Up to 3 3 lt DL 100 ppt 300 0 5 2 0 Each EDS Pak cartridge is delivered with a Certificate of Quality showing the analytical results of the Endocrine Disrupter Analysis performed on the cartridge lot Lit No PB1234ENUS Ordering information Description Catalogue No rer l l l EDS Pak Cartridge 1 pk delivered hermetically sealed with EDSPAKO01 a Certificate of Quality EDS Pak Cartridge installation and Conditioning Kit including the following reusable parts e Polyethylene 1 4 Gaz F Hose barb connection with O ring EDSKITOO1 e Polyethylene 1 4 G F 1 4 Gaz F connectors for connection to Millipak Note A 50 ml glass syringe is required to condition the EDS Pak cartridge with methanol This syringe is not provided by EMD Millipore Bibliography 1 Huang Y W Twidwell D L Elrod J C Occurrence and Effects of Endocrine Disrupting Chemicals in the Environment Practice Periodical of Hazardous Toxic and Radioactive Waste Management 7 4 p 241 247 2003 2 Brevini T A L Zanetto S B Cillo F Effects of En
13. f use ultrafiltration cartridge developed by EMD Millipore to meet the needs of researchers who require ultrapure water free of endocrine disrupters EDS The EDS Pak can be installed on all EMD Millipore Type water purification systems that deliver ultrapure water with TOC lt 5 ppb including the Milli Q Synergy Simplicity and Direct Q systems It is designed to provide at least 300 of EDS free ultrapure water before replacement Is necessary EDS Pak polisher specifications EDS Pak polisher operation The EDS Pak cartridge contains a specific type of activated carbon that has been validated for the removal of several endocrine disrupters The cartridge s housing and connector materials also have been validated for the absence of endocrine disrupters Before use the EDS Pak unit must be conditioned with methanol to ensure that all binding surfaces are accessible to endocrine disrupters Following this step the cartridge should then be flushed with ultrapure water to remove any excess methanol Once conditioned the EDS Pak unit is warranted to deliver up to 300 of EDSfree water see specifi cations below when fed by ultrapure water with a TOC level lt 5 ppb at a fl ow rate between 0 5 2 min Outlet Connection 1 4 Male and Luer F Inlet Connection 1 4 Male and Luer F Polyethylene Housing with O ring for easy Length 12 7 cm The EDS Pak has been validated for the
14. he EMD Millipore web site www millipore com labwater Benefits e The VOC Pak polisher is easily connected to the outlet of all EMD Millipore Type water purification systems Milli O Direct Q Synergy and Simplicity systems e The final purification step provides VOC free ultrapure water at high flow rate when you need it e Delivers a minimum of 300 of VOC free ultrapure water e Validated for efficient removal of VOCs e Each VOC Pak cartridge is delivered with a Certificate of Quality EMD MILLIPORE VOC Pak POLISHER DESIGN The VOC Pak cartridge is designed to assure scientists performing VOC analysis that the ultrapure water they use to rinse glassware produce standard solutions or dilute samples is VOC free Polypropylene Sintered Filter Inlet Connection 1 4 Outlet Connection 1 4 Male and Luer F Luer F Activated Carbon YY com GALLI Hose Connection with Polyethylene Housing O ring for easy water delivery Length 12 7 cm The purification media used to achieve this goal is a specific activated carbon selected after tests performed on media from many suppliers Table 1 Results of the VOC Pak challenge test for a processed volume of 350 amit a aR Challenge P tration after ppb op VOC Pak ppb benzene 0 97 lt 0 05 lt 0 05 bromodichloromethane 1 56 lt 0 05 lt 0 05 bromoform 1 58 lt 0 50 lt 0 50
15. highest eluted peak Specification of LC Pak ultrapure water ZRXQ003 WW At 210 nm lt 0 006 AU At 254 nm lt 0 002 AU Comments Concentration of 60 mL water at 1 mL min prior to elution No water preconcentration Optical properties absorbance in UV range Fluorescence as quinine UV 200 nm lt 0 05 AU UV 205 nm lt 0 01 AU UV 210 nm lt 0 01 AU At 254 nm lt 1 ppb At 365 nm lt 1 ppb UV 254 nm lt 0 005 AU Compliance with suitability for LC MS Reserpine test Residue after evaporation No peak higher than 10 ppb Reserpine at 609 m z in ESI lt 0 0001 w w Test performed as specified in ISO 3696 procedure An LC Pak validation guide is available should you wish to review the details of the validation report OPTIONAL Millipak 0 22 um end filter COMPARISON OF LC PAK AND SOME LC MS BOTTLED WATER In order to verify that fresh ultrapure water produced by a Milli OQ system and an LC Pak cartridge matched or exceeded the quality of commercially available bottled water for LC MS applications different 60 mL water Samples were pre concentrated on a C18 reverse phase chromatography column then eluted and an lyzed by LC MS PDA single quadrupole Examples of the results obtained are shown below UV plot at 210 nm Total plot ESI Total plot ESI From From m z 200 to 1000 m z 200 to 1000 6 2 4x10 280000 0 048 1 8x10 210000 gt 0 032 a 3 z W U gt E
16. ion limits ppb of VOCs analyzed in each lot of VOC Pak cartridges Additionally each lot of VOC Pak cartridges is analyzed to ensure that the following VOCs Table 2 are below the analytical quantification limit in water produced by a VOC Pak fed with ultrapure water from a EMD Millipore Milli O system resistivity at 18 2 MQ cm at 25 C and TOC lt 5 ppb Compound OL ppb benzene lt 0 05 bromobenzene lt 0 10 bromochloromethane lt 0 05 bromodichloromethane lt 0 05 bromoform lt 0 50 bromomethane lt 0 20 n butylbenzene lt 0 05 sec butylbenzene lt 0 50 tert butylbenzene lt 0 05 carbon tetrachloride lt 0 05 chlorobenzene lt 0 05 chloroethane lt 1 00 chloroform lt 0 05 2 chlorotoluene lt 0 05 4 chlorotoluene lt 0 10 dibromochloromethane lt 0 10 cance ne DBCP ae 1 2 dibromoethane lt 0 05 dibromomethane lt 0 10 1 2 dichlorobenzene lt 0 05 1 3 dichlorobenzene lt 0 05 1 4 dichlorobenzene lt 0 05 1 1 dichloroethane 0 05 1 2 dichloroethane lt 0 05 1 1 dichloroethene lt 0 05 cis 1 2 dichloroethene lt 0 05 trans 1 2 dichloroethene lt 0 05 1 2 dichloropropane lt 0 05 1 3 dichloropropane lt 0 10 Compound QL ppb 2 2 dichloropropane lt 0 05 1 1 dichloropropene lt 0 05 cis 1 3 dichloropropene lt 0 05 trans 1 3 dichloropropene lt 0 05 ethylbenzene lt 0 05 hexachlorobutadiene
17. lipak are registered trademarks of Merck KGaA Darmstadt Germany The M mark EMD Millipore are trademarks of Merck KGaA Waters Alliance XTerra Atlantis and Empower are registered trademarks of Waters Investment Ltd and ZQ is a trademark of Waters Investment Ltd Lit No DS1067ENUS 2012 EMD Millipore Billerica MA U S A All rights reserved EDS Pak Point of Use Polisher Endocrine disrupter free water at the ultrapure water point of delivery EMD MILLIPORE Key benefits e Point of use polisher for use with Milli Q and other EMD Millipore Type water purification systems e Final purification step provides endocrine disrupter free ultrapure water at high flow rate when you need it e Delivers a minimum of 300 of EDS free ultrapure water e Validated for efficient removal of bisphenol A diethyl phthalate and di n buty phthalate e Each EDS Pak cartridge is delivered with a Certificate of Quality EMD Millipore is a division of Merck KGaA Darmstadt Germany Endocrine disrupters Endocrine disrupters are small man made organic chemicals whose structures mimic those of hormones enabling them to interact with normal endocrine system functions By sheer coincidence their key shaped molecules open the hormonal locks that control proper development and behavior in both humans and animals Today endocrine disrupters are increasingly present in the environment Although endocrine disrupters m
18. ows easy production of ultrapure water that is both RNase free lt 0 01 ng mL and DNase free lt 4 pg uL Previous experiments also have shown that ultrafiltration with a properly validated device is just as efficient as diethylpyrocarbonate DEPC for RNase removal from ultrapure water without the negative aspects of DEPC treatment lengthy treatment time and contamination of the treated water by CO and ethanol BACTERIA FREEWATERPRODUCTION The validation protocol provided proof that the BioPak ultrafiltration cartridge enables delivery of bacteria free lt 1 cfu mL water when used according to Instructions with the filter outlet located in a clean environment BIOPAK ULTRA FILTRATION CARTRIDGE FLOW RATE Feed pressure 1 bar ma VU g9 _ LL 20 30 Water temperature C PRODUCT WATER SPECIFICATIONS Pyrogen Level EU mL lt 0 001 RNases ng mL lt 0 01 DNases pg L lt 4 Bacteria cfu ml lt 1 When used according to instructions with the filter outlet located in a clean environment HIGH FLOW RATE The large ultrafiltration membrane surface of the BioPak cartridge makes it possible to produce pyrogen free ultrapure water without compromising the flow rate as shown by the graph opposite The user can obtain ultrapure water on demand just before solution preparation minimizing risks of recontamination CERTIFICATE OF QUALITY Each BioP
19. titude that the ultrapure water that they are using in critical organic molecules analysis at trace and ultra trace levels will be adequate Polypropylene Compression Inlet connection 1 4 male Disc Sintered Filter and Luer F LLL C18 reverse phase silica 25 gra m Polypropylene Vyon Polypropylene Housing TUL Outlet connection 1 4 male Hose connection for and Luer F easy water delivery The granular C18 reverse phase silica in the polypropylene housing is tightly packed by a compression disc to avoid any channeling effect This ensures that the water passing through the LC Pak will be in close contact with the C18 chains binding organic molecules by hydrophobic interactions When required the LC Pak outlet can be connected to a 0 22 um in line filter LC PAK CARTRIDGE OPERATION The LC Pak is easy to use simply flush it with 50 mL methanol to properly wet the pores of the C18 reverse phase purification media connect the LC Pak to the outlet of the EMD Millipore ultrapure water system and flush with ultrapure water for 10 minutes You are now ready to deliver at least 500 L of ultrapure water with low trace organic LC PAK CARTRIDGE SPECIFICATIONS The LC Pak cartridge was extensively validated to ensure that the water quality at the outlet matches the most demanding requirements Parameter HPLC Gradient Test Absorbance of highest eluted peak HPLC Gradient Test Absorbance of
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