INSTRUCTION MANUAL FOR CHIRALPAK AY-RH, CHIRALCEL OX-RH and

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1 INSTRUCTIN MANUAL F CHIRALPAK AY-RH, X-RH ad Z-RH Please read this istructio sheet completely before usig these colums Colum Descriptio CHIRALPAK AY-RH X-RH Amylose tris(5-chloro-2-methylpheylcarbamate) coated o 5 µm Cellulose tris(4-chloro-3-methylpheylcarbamate) coated o 5µm R H 3 C H N Cl R HH N N CHCl 3 Cl CH 3 Z-RH Cellulose tris(3-chloro-4-methylpheylcarbamate) coated o 5 µm R H N Cl CH 3 Page 1

2 CAUTIN The etire HPLC system icludig the ijector ad the ijectio loop must be flushed with a solvet compatible with the colum ad its storage solvet prior to coectig the colum. May of the solvets commoly used i HPLC eluets such as acetoe, chloroform, DMF, dimethylsulfoxide, ethyl acetate, methylee chloride ad THF may DESTRY the chiral statioary phase if they are preset, eve i residual quatities, i the system. If a auto-sampler is used, the the solvet employed to flush this uit betwee ijectios should also be chaged ad the relevat solvet lies flushed. peratig Coditios Flow rate directio 150 x 2.1 mm i.d. Aalytical colum As idicated o the colum label 150 x 4.6 mm i.d. Aalytical colum Typical Flow rate ~ 0.1 to 0.3ml/mi ~ 0.5 to 1.0ml/mi Pressure limitatio Should be maitaied < 300 Bar (4350 psi) for maximum colum life Adapt flow rates to colum size. ph Betwee ad ph 9.0 Temperature 5 C to 40 C A ph less tha 8.0 is recommeded for maximum colum life. Keep the temperature betwee 5 C to 25 C whe used with a ph higher tha 7.0 peratig Procedure Please cotact Chiral Techologies for further assistace before tryig ay solvets ot metioed below. A - Mobile Phases / For UV ad Mass Detectios ACIDIC (AMPHTERIC) NEUTRAL BASIC CHIRALPAK AY-RH X-RH Z-RH Aqueous solutio rgaic modifier HCH aq. Water CH 3CN or MeH or EtH or IPA 20mM NH 4HC 3 aq. ph 9.0 adjusted with a basic additive Typical startig coditios Aqueous solutios 60% CH 3CN 40% NTE 1: If you caot achieve sufficiet resolutio, try the complemetary mobile phases: Page 2

3 B Complemetary Mobile Phases / For UV Detectio ACIDIC (AMPHTERIC) NEUTRAL BASIC CHIRALPAK AY-RH X-RH Z-RH Aqueous solutio 50mM Phosphate Buffer H 3P 4 aq. 100mM KPF 6 (or NaPF 6) aq. adjusted with H 3P 4 Water 20mM Borate Buffer ph mM Phosphate Buffer ph mM KPF 6 (or NaPF 6) aq. rgaic modifier Typical startig coditios CH 3CN or MeH or EtH or IPA Aqueous solutios 60% CH 3CN 40% NTE 2: The cocetratio of all the bufferig salt should be less tha 500mM. Refer to sectio C for preparatio of aqueous solutio ad choice of basic additives. It is recommeded to use CH 3 CN to start the ivestigatio. The elutio power of orgaic modifiers for these colums is i the descedig order of CH 3 CN > EtH > MeH: 50%CH 3 CN 65-70%EtH 75-80%MeH. The use of other orgaic solvets has ot bee ivestigated ad could be harmful to the colums. The use of alcohols causes the back pressure to be sigificatly higher compared to CH 3 CN due to their high viscosity i mixtures with water. Retetio ca be adjusted by chagig the proportio of CH 3 CN. Retetio may be very sesitive to the amout of CH 3 CN preset ito the mobile phase. Lowerig the colum temperature may icrease the retetio time ad the selectivity. Icreasig the colum temperature ad decreasig the flow rate may icrease the resolutio. To maximize colum life the use of a guard cartridge is essetial whe basic coditios are employed. The use of strog basic coditios (> ph 9) must be avoided, as they are kow to damage the silica gel matrix. Whe these colums are used at ph > 7, the temperature should be maitaied betwee 5 C ad 25ºC for maximum colum life. High percetages of orgaic modifier i the mobile phase may precipitate the bufferig salt from the solutio, ad lead to cosequet cloggig of the colum (refer to the table below). Water / rgaic Modifier Buffer solutio / rgaic Modifier 90 / 10 to 0 / / 10 to 15 / 85 Do ot use the phosphate buffer for ph > 8. Whe ph 9 is ecessary, use the ammoium bicarboate solutio or borate buffer for maximum colum life. Page 3

4 C Buffer preparatio Examples Preparatio of ph 2 Phosphate buffer: Solutio A: 50mM potassium dihydrogephosphate 3.40g KH 2P 4 / FW , make up the volume to 500ml with HPLC grade water Solutio B: phosphoric acid (H 3P 4 85% by weight) Adjust the ph of solutio A to a value of 2.0 usig solutio B. Preparatio of ph 2 KPF 6 (NaPF 6) solutio: Solutio A: 100mM potassium (sodium) hexafluorophosphate 9.20g KPF 6 / FW or 8.40g NaPF 6 / FW , make up the volume to 500ml with HPLC grade water Solutio B: phosphoric acid (H 3P 4 85% by weight) Adjust the ph of solutio A to a value of 2.0 usig solutio B. Preparatio of ph 9 Ammoium bicarboate solutio: Solutio A: 20mM ammoium bicarboate 0.78g NH 4HC 3 / FW 78.05, make up the volume to 500ml with HPLC grade water Solutio B Basic additive such as diethylamie (DEA), triethylamie (TEA), ammoia (NH 3) ad so o. * DEA teds to give better peak shape tha other bases. Adjust the ph of solutio A to a value of 9.0 usig solutio B. Preparatio of ph 8 Phosphate buffer: Solutio A: 20mM potassium hydrogeophosphate 1.74g of K 2HP 4 / FW , make up the volume to 500ml with HPLC grade water Solutio B: 20mM potassium dihydrogeophosphate 1.36g KH 2P 4 / FW , make up the volume to 500ml with HPLC grade water. Adjust the ph of solutio A to a value of 8.0 usig solutio B. Preparatio of ph 9 Borate buffer: Solutio A: 20mM sodium tetraborate decahydrate 3.81g of Na 2B H 2 / FW , make up the volume to 500ml with HPLC grade water Solutio B: 20mM boric acid 0.62g H 3B 3 / FW 61.83, make up the volume to 500ml with HPLC grade water Adjust the ph of solutio A to a value of 9.0 usig solutio B. Colum Care / Maiteace The use of a guard cartridge is highly recommeded for maximum colum life. Samples should preferably be dissolved i the mobile phase. The mobile phase ad the sample solutio should be filtered through a membrae filter of approximately 0.5µm porosity to esure that there is o precipitate before usig. Before discoectig the colum from the HPLC, ay traces of salts should be removed by flushig with a mobile phase that does ot cotai ay salts / buffers, e.g. Water/CH 3 CN 60:40 (v/v). If the colum is cotamiated with o eluted compoets, wash it with a mobile phase that does ot cotai ay salts / buffers the with 100% CH 3 CN for 2 hours at 0.3ml/mi. If the o-elutig compoets are more soluble i methaol, this solvet may be used for the washig step. All salts must be flushed out from the HPLC system ad the colum before chagig to 100% CH 3 CN or 100% methaol. Use Water/CH 3 CN 60:40 (v/v) to store the colum. Page 4

5 Importat Notice STRNGLY BASIC solvet additives or sample solutios MUST BE AVIDED, because they are likely to damage the silica gel used i these colums. peratig these colums i accordace with the guidelies outlied here will result i a log colum life. I the USA: questios@chiraltech.com or call CHIRAL I the EU: cte@chiral.fr or call +33 (0) I Idia: chiral@chiral.daicel.com or call Locatios: North/Lati America Chiral Techologies. Ic. 800 North Five Poits Road West Chester, PA CHIRAL Tel: Fax: chiral@chiraltech.com Europe Chiral Techologies Europe Parc d Iovatio Bd Gothier d Aderach Illkirch Cedex, Frace Tel: Fax: cte@chiral.fr Idia Daicel Chiral Techologies (Idia) Pvt. Ltd. Lab No. 4A, Phase III IKP Kowledge Park Geome Valley, Turkapally, Shameerpet, Raga Reddy Dist. Hyderabad , Telagaa Tel: Fax: chiral@chiral.daicel.com CHIRALCEL, CHIRALPAK ad CRWNPAK are registered trademarks of DAICEL CPATIN Page 5

INSTRUCTION MANUAL FOR CHIRALPAK ZWIX(+) and CHIRALPAK ZWIX(-)

INSTRUCTION MANUAL FOR CHIRALPAK ZWIX(+) and CHIRALPAK ZWIX(-) 800 orth Five Points Road West Chester, PA 19380 800 6 CIRAL Tel: 610-594-2100 Fax: 610-594-2325 chiral@chiraltech.com www.chiraltech.com ISTRUCTI MAUAL FR CIRALPAK ZWIX(+) and CIRALPAK ZWIX(-) Semi-Preparative