Page 1 of 2 CAPCELL PAK C18 MGIII Type 13.02.2014 http://hplc
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1. hplc shiseido co jp e column html mg3_ index htm 13 02 2014 CAPCELL PAK C18 MGIII Type Page 2 of 2 groups on the stationary phase surface For the former a special method to clean and Before PC condition columns pre conditioning PC was CHa After PC invented The PC process was intensively Si O studied and one of the results was shown in Fig 2 where the relation of the duration time of PC and tr of amitriptyline was clearly observed The retention time of naphthalene a conventional indicator for overall retention of Cis columns did not change at all while tr of amitriptyline gradually increased with the PC time and reached the plateau after a certain time Solid state Si NMR a useful tool to directly observe different silanol groups on the surface 10 20 30 40 50 60 70 8 0 100 110 120 130 PP did not show a significant difference between before and after the PC process Fig 3 which Fig 3 Spectral difference in 7 Si NMR suggested the residues of unwanted substances were the most responsible to the tr variation of amitriptyline Based on these results we confidently applied the PC process to all of the MG columns and decided to launch the columns as MGIII series The followings are some of characteristic applications obtained with the MGIH series Therefore the special PC process provided the stable retention to both of basic amitriptyline and neutral naphthalene Ref 1 J J Kirkland cow2. 0 90 1 40 1 26 As Amitriptyline 7 0 90 1 60 1 42 k Amitriptyline i 12 17 1 3 See Fig 1 on reverse i Chromatographic conditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7uL Detector UV 254 nm Temperature 40 C 5 See Fig 2 on reverse Chromatographic conditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2POz 20 mmol L K2HPO Water Methanol 20 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C T See Fig 3 on reverse Acid Resistance gt 80 0 86 3 Alkali Resistance gt 70 0 T95 Chromatographic conditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm Percentage of k at 20 hr over the initial k 8 Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C 9 Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGI S 3 Lot BMSII03 0 4 8 12 min Fig 1 Alkyibenzene 0 3 6 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm D x 100 mm Mobile phase Acetonitrile Water 30 70 viv Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume
3. provide better column lifetimes 8 To prevent column deterioration avoid the following Frequent change of the mobile phase composition Rapid change in pressure of column inlet Continued using at pressure exceeding 15 MPa 100 MPa for IF2 40 MPa for IF and 50 MPa for MGIII H High column pressure due to the use of a high viscosity mobile phase Prolonged water flow 3 2 Preparing a Sample Solution 1 Dissolve the sample in a solvent of the same composition as the eluate wherever possible 2 Using a solvent with strong dissolving power may lower the separation efficiency or cause the sample to precipitate at the column head 3 If there is insoluble matter remaining in the sample solution filtrate the solution using a filter 0 45 um or smaller 4 The pH of the sample solution should be set in acceptable pH range for packing material 3 3 Notes on Analysis Columns of CAPCELL PAK series generally show similar separation profiles to those of corresponding conventional silica based columns although slight selectivity difference may be observed depending on the analyte When optimizing the conditions for compounds already done with conventional columns use the same condition as a starting one 1 When using C1s Cs Phenyl C1 or CN 1 Regarding the guard column Use a column of the same packing material as the main column If a guard column of different packing material or the chemically bonded silica column of a different manufact
4. 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene 0 3 6 min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Methanol H20 Formic Acid 500 500 1 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 3 0p1L Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BMSII02 the previous lot CAPCELL PAK MG IW Certificate of Analysis CAPCELL PAK C18 MGI S 3 Lot BMSI04 Analysis of Silica gel Specification Result Mean Particle Diameter um 26 3 2 2 9 Particle Distribution 40 90 lt 1 28 1 24 Multi point Nitrogen Sorption Median Pore Diameter nm 80 40 0 8 8 Surface Area m g 250 310 295 Pore Volume mL g 0 80 1 00 0 87 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 2 6 Zn lt 5 0 n d Mg lt 5 0 n d Ti lt 5 0 0 3 Al lt 5 0 0 5 m Na lt 5 0 0 3 ICP AES n d not detected Analysis of CAPCELL PAK C18 MGI S 3 Carbon Content 13 4 15 5 14 6 Chromatographic Results a Hexylbenzene Amylbenzene E 1 46 1 50 1 49 k Hexylbenzene 11 3 15 2
5. Water 70 30 v v Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene 0 3 6 min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Methanol H20 Formic Acid 500 500 1 viv Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 3 0L Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BMSII01 the previous lot CAPCELL PAK MG QW Certificate of Analysis CAPCELL PAK C18 MGI S 3 Lot BMS M03 Analysis of Silica gel Specification Result Mean Particle Diameter um 26 32 2 8 Particle Distribution 40 90 lt 1 28 1 22 Multi point Nitrogen Sorption Median Pore Diameter nm 8 0 10 0 8 6 Surface Area m g 250 310 298 Pore Volume mL g 0 80 1 00 0 86 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 3 0 Zn lt 5 0 n d Mg lt 5 0 n d Ti lt 5 0 n d Al lt 5 0 1 3 Na lt 5 0 n d ICPIAES n d not detected Analysis of CAPCELL PAK C18 MGI S 3 Carbon Content 13 4 155 14 6 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 49 k Hexylbenzene 11 3 452 13 0 a Methyl benzoate Toluene 0 38 0 47 0 42 As Pyridine 0 90 1 50 1 21 As Phenol 0 90 1 20 1 01 As Quinizarin
6. conditions Inadequate column equilibration time Page 4 of 4 e Sonicate the filter or replace it e Filtrate the mobile phase and sample solution in advance using a membrane filter e Attach a line filter e Clean the tubing and replace the plunger seal e Prepare a sample solution with the mobile phase Reconnect the tubing Review the pH salt concentration sample amount and other conditions Review the ion pair agent concentration pH sample amount and other conditions eCheck the column performance using standard inspection solution e Check the pump and tubing for any leaks e Secure adequate equilibration time e Secure adequate equilibration time CAPCELL PAK is shipped after a strict performance check However if you should find any defect please contact your dealer or Shiseido for replacement Note that Shiseido does not warrant the product against column life or deterioration caused by the failure to follow the above handling instructions Ten or more days after reception by the customer Shiseido will assume that the product was delivered in good condition and will not accept a later replacement request Our Chromatography Business http hplc shiseido co jp e column html cp_manual htm 2008 10 1 2nd edited 2011 8 1 a PAGE TOP m Contact Shiseido Site Map 13 02 2014 GAP GE LL PAK MAGIE Certificate or Analysis CAPCELL PAK C18 MGIII S 5 Lot BSII03 Analysis of Sili
7. or more to equilibrate the column Since the equilibrating time depends on the flow volume and the salt concentration allow the mobile phase to remain at a high flow volume or the same pH and high salt concentration in urgent cases Ascorbic acid may show peak tailing Allantoin does not ionize in the pH range from 2 to 8 However the use of a phosphate buffer solution is advised Example of mobile phase Acetonitrile water 80 20 5 mmol L KH2P04 pH 2 0 H3PO4 3 When using CR Packing material of CR is a mixture of C18 and SCX In ion exchange mode the following factors will change elution behavior significantly pH pH should ideally be 2 0 or more away from pKa for full ion separation from the sample Salt concentration Amount of organic solvent http hplc shiseido co jp e column html cp_manual htm 13 02 2014 CAPCELL PAK User s Manual Page 3 of 4 Salt type 4 Storing the Column 1 Seal the column with the accessory plug and store it in a cold place where there is little temperature fluctuation 2 Replace the column with solution of organic solvent and water having the same composition as the mobile phase after using solvents containing strong organic acids such as TFA or basic solvents Do not use 100 water Moreover for storage of one week or longer replace the column with acetonitrile 3 For storage within one month after using replace the mobile phase with a solution of organic solvent and water hav
8. x 150 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm Percentage of k at 20 hr over the initial k ee phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGIII S 5 Lot BSIII03 2 3 4 5 IA o 5 10 15 20 25 min Fig 1 Alkylbenzene m 1 2 Es at L di as 0 5 10 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm D x 150 mm Mobile phase Acetonitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Methanol H20 Formic Acid 500 500 1 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Amitrip
9. 12 8 a Methyl benzoate Toluene 0 38 0 47 0 42 As Pyridine 0 90 1 50 1 23 As Phenol 0 90 1 20 1 01 As Quinizarin 0 90 1 40 1 13 As Amitriptyline 7 0 90 1 60 1 52 k pal is 12 17 1 2 eee Fig 1 on reverse 4 Chromatographic conditions Column 4 6 mm I D x 100mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7pL Detector UV 254 nm eee 40 C 5 See Fig 2 on reverse Chromatographic conditions Column 4 6 mm I D x 100mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2PO4 20 mmol L KzHPO Water Methanol 20 80 v v Injection volume 5yL Detector UV 254 nm Temperature 40 C 7 See Fig 3 on reverse Acid Resistance gt 80 0 85 6 Alkali Resistance gt 70 0 71 2 Chromatographic conditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7pL Detector UV 254 nm Percentage of k at 20 hr over the initial k Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGI S 3 Lot BMSI104 0 4 8 12 min Fig 1 Alkylbenzene min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm 1 D x 100 mm Mobile phase Aceto
10. 4 6 mm 2 pieces 21107 End fitting 6 mm 2 pieces 21110 Ferrule 1 16 Ferrules 1 16 10pieces a PAGE TO 7 Troubleshooting Problems in high performance liquid chromatography are attributable to various causes that cannot all be listed up The table below describes some comparatively common problems related to the column Symptom Cause Measures http hplc shiseido co jp e column html cp_manual htm 13 02 2014 CAPCELL PAK User s Manual 1 Column pressure rise 2 Peak splitting tailing and broadening 3 Retention time too long or unstable 4 Retention time too short Blocking with foreign matter 1 Dust or insoluble matter in the mobile phase or sample solution 2 Dirt in the tubing 3 4 Precipitation of sample Plunger seal fragment components Void in the column head Dead volume due to inappropriate connections Inappropriate mobile phase conditions Ion suppression method Inadequate suppression Too much sample Ion pair method Inadequate concentration of the ion pair agent Too much sample Column deterioration Not repairable in the case of column deterioration or damage to the packing condition Liquid leak Indicated on the pressure gage of the pump Inappropriate mobile phase conditions Inadequate column equilibration time Hydrolysis deterioration of a bonded groups by strong acid or base Inappropriate mobile phase
11. 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C T See Fig 3 on reverse Acid Resistance gt 80 0 86 7 Alkali Resistance gt 70 0 74 9 Chromatographic conditions Column 4 6 mm I D x150 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7L Detector UV 254 nm Percentage of k at 20 hr over the initial k 8 Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information CAPCELL PAK C18 MGI S 5 Lot BS 04 0 5 10 15 20 25 min Fig 1 Alkylbenzene 0 5 10 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Acetonitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm D x 150 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Methanol H20 Formic Acid 500 500 1 v v Flow rate 1 0 mL min Temperat
12. CAPCELL PAK C18 MGIII Type Page 1 of 2 SHISEIDO Products CAPCELL PAK Cts MGIII Type Flist of Sales 2 Application Data _ Representatives Overview User s Manual Validation Sheet Contact Shiseido Catalog List About MGIII The third generation of the MG series was developed to overcome the Column to column variation in retention of basic compounds under an acidic condition We hope that the quality of MGIII will help develop improved methods in various LC MS applications Hydrophobicity and surface polarity parameters of popular C18 columns Intended to be the standard for LC 0 7 MS While it is based on the identical silica 0 65 e l CHUGIZ0 used in other MG series CAPCELL 0 6 PAK Cis MGIII possesses a weaker cuG120 hydropobicity and a stronger surface ga 9 55 PhuUGIZ0 polarity and is suitable for a wide a as CaUGIZ0 Gang atlantis variety of applications such as analyses gs CaDD ota wae of metabolites and other highly polar as Inertsil 005 3 compounds Especially its a WOES ypl column reproducibility in retention times of CiaSG120 HC Pro p baji ne 0A IF basic compounds under acidic conditions CauGizoy SUPERIOREX i CiBACR CalGeo is a preferred feature for long time span 0 35 Symmetry CR applications such as clinical analyses with LC MS and applications under 1 7 1 8 1 9 2 24 22 GLP MGIII s specifications include a n separation performance of amitriptyline droph fogs
13. Mobile phase Acetonitrile Water Mobile phase 30 70 v v Methanol H20 Formic Acid Flow rate 1 0 mL min 00 500 1 v v Temperature 40 C Flow rate 1 0 mL min Detector UV 254 nm Temperature 40 C Injection volume 7 0uL Detector UV 254 nm Sample 1 Pyridine injection volume 5 0uL 2 Phenol Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BSII 05 the previous lot J EIDO CAPCELL PAK MGI Certificate of Analysis CAPCELL PAK C18 MGI S 3 Lot BMSMO01 Analysis of Silica gel Specification Result Mean Particle Diameter um 26 32 2 8 Particle Distribution 40 90 lt 1 28 1 22 Multi point Nitrogen Sorption Median Pore Diameter nm 8 0 10 0 8 6 Surface Area m g 250 310 298 Pore Volume mL g 0 80 1 00 0 86 7 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 3 0 Zn lt 5 0 n d Mg lt 5 0 n d Ti lt 5 0 n d Al lt 5 0 1 3 Na lt 5 0 n d 2 ICPIAES n d not detected Analysis of CAPCELL PAK C18 MGI S 3 Carbon Content 13 4 155 14 5 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 49 k Hexylbenzene 11 3 15 2 13 8 a Methyl benzoate Toluene 0 38 0 47 0 42 As Pyridine 0 90 1 50 1 45 As Phenol 0 90 1 20 1 19 As Quinizarin 0 90 1 40 1 09 As Amitriptyline 0 90 1 60 1 30 k T H 12 4 7 1 6 gece Fig 1 on reverse 4 Chromatographic conditions Column 4 6 mm I D
14. ca gel Specification Result Mean Particle Diameter um 440 470 4 66 Particle Distribution 40 90 lt 1 28 1 24 Multi point Nitrogen Sorption Median Pore Diameter nm 9 0 11 0 9 8 Surface Area mg 230 280 263 Pore Volume mL g 0 85 0 99 0 93 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 2 0 Zn lt 5 0 n d Mg lt 50 n d Ti lt 50 n d Al lt 50 2 0 Na lt 50 2 5 2 ICP AES n d not detected Analysis of CAPCELL PAK C18 MGIII S 5 Carbon Content 140 155 14 5 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 48 k Hexylbenzene 11 3 15 2 12 9 a Methyl benzoate Toluene 0 42 0 46 0 44 As Pyridine 0 90 1 50 1 01 As Phenol 0 90 1 20 1 00 As Quinizarin 0 90 1 40 1 02 As Amitriptyline 7 0 90 1 30 1 13 k Amitriptyline 7 13 16 1 4 wee Fig 1 on reverse i Chromatographic conditions Column 4 6 mm I D x 150mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 viv Injection volume 7uL Detector UV 254 nm omperature 40 C lt See Fig 2 on reverse Chromatographic conditions Column 4 6 mm I D x150 mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2POz 20 mmol L K2ZHPO Water Methanol 20 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C 7 See Fig 3 on reverse Acid Resistance gt 80 0 83 8 Alkali Resistance gt 70 0 79 3 Chromatographic conditions Column 4 6 mm I D
15. ction volume 7pL Detector UV 254 nm ig Temperature 40 C 5 See Fig 2 on reverse Chromatographic conditions Column 4 6 mmI1 D x150mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2PO 20 mmol L K2HPO Water Methanol 20 80 v v injection volume 5pL Detector UV 254 nm Temperature 40 C 7 See Fig 3 on reverse Acid Resistance gt 80 0 83 7 Alkali Resistance gt 70 0 73 7 Chromatographic conditions Column 4 6 mm 1 D x150 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm Percentage of k at 20 hr over the initial k Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C 9 Mobile phase 4 mmol L Sodium Tetraboraie Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGI S 5 Lot BSI06 Chromatographic conditions Column 4 6 mm D x 150 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 ml min Temperature 35 C Detector UV 254 nm Injection volume 5 0L Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene min Fig 1 Alkylbenzene 0 5 10 0 5 10 min min Fig 2 Pyridine Phenol Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Chromatographic conditions Column 4 6 mm I D x 150 mm Column 4 6 mm I D x 150 mm
16. ecrease It will vary depending upon the temperature and the concentration of organic solvent For continued high performance of the columns ensure to avoid using pH exceeding the recommended ones In addition high temperature and organic solvent poor condition will result in short column lifetimes when working at the extremes of pH Table 1 Applicable pH range for capcell pak columns Bonding group Cis Cs Phenyl C1 CN NH2 C18 SCX UG UG UG IF Type ACR MG AQ DD AG SG SG CR SG AG Applicable pH 1 10 2 10 2 9 1 5 10 2 10 2 9 2 8 2 7 Note The durability of C1 and CN column are not comparable with that of Cs and Phenyl columns because of their short function groups http hplc shiseido co jp e column html cp_manual htm 13 02 2014 CAPCELL PAK User s Manual Page 2 of 4 3 After full degassing filtrate the mobile phase using a membrane filter 0 45 um or smaller to remove dust 4 A 2 um filter is used at the column inlet To prevent foreign matter from clogging the column inlet filter we recommend using a line filter 5 The mobile phase stated in the report is sealed in a new column To change to a mobile phase containing inorganic salt note the replacement procedure to avoid the salting out 6 Ion pair reagent will slightly result in short column lifetimes 7 AQ column is applicable 100 aqueous mobile phase However the column lifetime will extremely vary depending upon the HPLC conditions It is known that acidic phosphate buffer will
17. ide Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Methanol H20 Formic Acid 500 500 1 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BS11104 the previous lot SHISEIDO CAPCELL PAK MGH Certificate of Analysis CAPCELL PAK C18 MGI S 5 Lot BSM 06 Analysis of Silica gel Specification Result Mean Particle Diameter um 440 470 4 55 Particle Distribution 40 90 lt 1 28 1 23 Multi point Nitrogen Sorption Median Pore Diameter nm 9 0 11 0 9 8 Surface Area Im g 230 280 255 Pore Volume mL g 0 85 0 99 0 90 i Quantachrome Autosorb 14 Metal Contents ppm Fe lt 50 0 8 Zn lt 5 0 n d Mg lt 5 0 n d Ti lt 50 0 5 Al lt 5 0 25 Na lt 5 0 n d ICP AES n d not detected Analysis of CAPCELL PAK C18 MGIILS 5 Carbon Content 140 15 5 14 2 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 48 k Hexylbenzene 114 3 15 2 12 3 a Methyl benzoate Toluene 0 42 0 46 0 43 As Pyridine 0 90 1 50 0 98 As Phenol 0 90 1 20 0 95 As Quinizarin 0 90 1 40 1 16 As Amitriptyline 7 0 90 1 30 1 12 k Amitriptyline 7 13 16 1 3 3 See Fig 1 on reverse Chromatographic conditions Column 4 6 mm I D x 750mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Inje
18. in Mobile phase 20 mmol L KH2PQ4 20 mmol L K2HPO Water Methanol 20 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C See Fig 3 on reverse Acid Resistance gt 80 0 86 2 Alkali Resistance gt 70 0 76 7 Chromatographic conditions Column 4 6 mm I D x150 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm Percentage of k at 20 hr over the initial k 8 Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGM S 5 Lot BS105 0 5 10 15 20 25 min Fig 1 Alkylbenzene 0 5 10 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm I D x 150 mm Mobile phase Acetonitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm 1 D x 150 mm Mobile phase Acetonitrile Water 70 30 viv Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene Fig 3 Amitriptyline Hydrochlor
19. ing the same composition as the mobile phase and then fill it with the solvent used at the time of shipment Refer to the column report 4 Avoid using 100 water to rinse a column that is other than AQ 5 End Fittings 1 An analytical column of up to 6 mm ID uses a filter embedded end fitting as shown in Fig 1 The filter cannot be changed alone If the filter is clogged or the column pressure is high replace the end fitting See Table 3 for the replacement parts and repair items 2 See Fig 1 for the column connection If the tubing is inappropriate especially if a tube for a different type of column is used the length after the ferrule tip V in Fig 1 is often different from the end fitting length L and a problem may occur If L is greater than V dead volume may be generated and cause peak broadening or tailing or deterioration of separation performance If L is smaller than V liquid may leak because of inadequate ferrule adhesion Therefore we recommend replacing the ferrule together with the column If the column is replaced frequently the male nut may crush the ferrule and liquid may leak Since tightening the nut too much may cause its head to come off replace the ferrule at an early stage End fitting Ferrule 1 16 Male nut Tube 1 16 Fig 1 Column connection amp PAGE TC 6 Replacement Parts and Repair Items Table 3 Replacement parts and repair items Part No Part Name Description 21105 End fitting
20. nia iydropnonisily under an acidic condition Features HPLC Conditions Column Column A 4 6 mm i d x 150 mm Mobile phase CH30H H20 HCOOH 500 500 1 Flow rate 1 mL min Temperature 40 C Detection UV 254 nm Injvol 5 wb Sample dissolved in H20 50 g mL Fig 1 Example of lot variation under an acidic condition An example of column to column variation under an acidic condition is shown in Figure 1 The compound used here was amitriptyline a highly basic compound used for the USP evaluation method The results were obtained under an isocratic condition 0 1 formic acid Es i ra Fit L methanol Retention times of the three p E an llts columns Column A anonymous were found io very different under the mobile phase that was HI ar i a Pre conditioning Stable retention is reached one of the most common in LC MS The similar tendency was also found in many other columns Fig 2 Optimization of pre conditioning time We carried out a comprehensive study to understand the phenomenon of tR variation including effects of metal impurities reagent grade of mobile phase solvents and byproducts in the process of stationary phase synthesis After eliminating one possible cause after another we reached a conclusion that the responsible factor was either 1 the synthetic byproducts and the residues of impurities from reagents used for the synthetic process or 2 isolated silanol http
21. nitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0L Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 mL min Temperature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm 1 D x 100 mm Mobile phase Methanol H20 Formic Acid 500 500 1 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 3 0uL Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BMSI103 the previous lot
22. onditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7uL Detector UV 254 nm Temperature 40 C 5 See Fig 2 on reverse Chromatographic conditions Column 4 6 mm 1 D x 100mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2PO4 20 mmol L K2HPO Water Methanol 20 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C 7 See Fig 3 on reverse Acid Resistance gt 80 0 85 5 Alkali Resistance gt 70 0 77 6 Chromatographic conditions Column 4 6 mm I D x100 mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm Percentage of k at 20 hr over the initial k gle phase Methanol Water TFA 10 89 14 pH 1 0 Temperature 60 C 2 Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGI S 3 Lot BMSI02 0 gt 4 8 12 16 min Fig 1 Alkylbenzene 0 3 6 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Acetonitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm D x 100 mm Mobile phase Acetonitrile
23. orkers J Chromatogr 352 1986 275 Excellent reproducibility gt Fig 4 Fig 5 Comparison of column bleeding gt Fig 6 MEID Separation of organic acids Fig 8 Also available worldwidely GLP GMP Supporting column Categorized as L1 in USP Property values Particle Specific P D Pore size size Surface area C ensity Functional group Acceptable USP nm um m g umol m pH 10 3 300 15 2 3 Octadecyl group 2 10 L1 10 5 260 15 2 7 Octadecyl group 2 10 L1 PAGE TOP Our Chromatography Business Contact Shiseido Site Map MZ Analysentechnik GmbH _ E E I W hlerstra e 2 6 D 55120 Mainz Tel 49 6131 68 66 19 ANALYSENTECHNIK Fax 49 6131 68 66 20 e mail info mz at de www mz at de http hplc shiseido co jp e column html mg3_ index htm 13 02 2014 CAPCELL PAK User s Manual Page 1 of 4 SHISEIDO Products CAPCELL PAK User s Manual El Microcotumn User s Manual MG Series MGI MGT MG UG SG AG CR UHPLC columns IF2 IF MGIII H To Top of HPLC Columns E List of Sales Technical Materials Representatives Contact Shiseido Catalog List 1 Handling the Column 2 Attaching the Column Analysis 4 Storing the Column 5 End Fittings 6 Replacement Parts and Repair Items 7 Troubleshooting Le i Le Le Lo Lt Le CAPCELL is provided with packing material made of totally porous spherical silica coated with a mono layer silicone polymer having octadecy C18 as well as other f
24. perature 35 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Propylbenzene 3 Butylbenzene 4 Amylbenzene 5 Hexylbenzene 0 3 6 min Fig 3 Amitriptyline Hydrochloride Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Methanol H20 Formic Acid 500 500 1 viv Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 3 0uL Sample 1 Uracil 2 Amitriptyline hydrochloride IJ EIDO capceLtt PAK maem Certificate oF Analysis CAPCELL PAK C18 MGI S 3 Lot BMS M02 Analysis of Silica gel Specification Result Mean Particle Diameter um 26 32 2 8 Particle Distribution 40 90 lt 1 28 1 21 Multi point Nitrogen Sorption Median Pore Diameter nm 8 0 10 0 8 6 Surface Area m g 250 310 291 Pore Volume mL g 0 80 1 00 0 85 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 2 8 Zn lt 5 0 0 3 Mg lt 5 0 n d Ti lt 5 0 0 5 Al lt 5 0 2 8 Na lt 5 0 n d 2 ICP AES n d not detected Analysis of CAPCELL PAK C18 MGI S 3 Carbon Content 13 4 15 5 14 2 Chromatographic Results a Hexylbenzene Amylbenzene 146 1 50 1 49 k Hexylbenzene 11 3 15 2 13 7 a Methyl benzoate Toluene 0 38 0 47 0 42 As Pyridine 0 90 1 50 1 45 As Phenol 0 90 1 20 1 18 As Quinizarin 0 90 1 40 1 03 As Amitriptyline 7 0 90 1 60 1 34 k Amitriptyline mt 12 17 1 3 See Fig 1 on reverse i Chromatographic c
25. tyline hydrochloride The chromatogram shown with a dotted line is that of Lot BSIII02 the previous lot CAPCELL PAK MG I Certificate of Analysis CAPCELL PAK C18 MGI S 5 Lot BS 04 Analysis of Silica gel Specification Result Mean Particle Diameter um 4 40 4 70 4 54 Particle Distribution 40 90 lt 1 28 1 24 Multi point Nitrogen Sorption Median Pore Diameter nm 9 0 11 0 9 9 Surface Area m7 g 230 280 260 Pore Volume mL g 0 85 0 99 0 92 1 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 2 2 Zn lt 5 0 0 5 Mg lt 5 0 n d Ti lt 5 0 n d Al lt 5 0 2 2 _ Na lt 5 0 2 5 ICP AES n d not detected Analysis of CAPCELL PAK C18 MGIII S 5 Carbon Content 14 0 15 5 14 3 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 48 k Hexylbenzene 11 3 15 2 13 1 a Methyl benzoate Toluene 0 42 0 46 0 43 As Pyridine 0 90 1 50 1 12 As Phenol 0 90 1 20 1 01 As Quinizarin 0 90 1 40 1 01 As Amitriptyline 7 0 90 1 30 1 13 k pai dite Pe 13 16 1 3 4566 Fig 1 on reverse 4 Chromatographic conditions Column 4 6 mm i D x150mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7pL Detector UV 254 nm lemperature 40 C See Fig 2 on reverse 6 Chromatographic conditions Column 4 6mm I D x150 mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2PO4 20 mmol L K2HPO Water Methanol 20
26. unctional groups The epoch making packing material integrates the high separation performance and pressure resistance of silica packing material and the durability of organic polymer based packing material 1 Handling the Column 1 Handle the column with great care A strong shock may cause damage 2 Attach or detach the column when the pressure gage indicates zero 3 The maximum column operating pressure is 100 MPa for IF2 50 MPa for MGIII H 40 MPa for IF and 20 MPa for others 2 Attaching the Column 1 The column joint is of the male nut type for tubing of 1 16 inch OD Check that the tubing joints of the system fit correctly and that the ferrule tips are deeply inserted into the joints See Fig 1 2 Before attaching the column replace the liquid in the system with the mobile phase to be used Note the replacement procedure to avoid salting out The shipment solvent is described in the column report enclosed with the column 3 Attach the column according to the direction of the arrow 3 Analysis 3 1 Mobile Phase 1 All solvents acceptable for the conventional chemically bonded silica columns can be used 2 The applicable pH range for capsule type packing material depends on the bonding group The recommended operating pH ranges for Capcell Pak columns are listed in Table 1 The applicable range of pH is determined by operating pH limits under which the retention and the number of theoretical plate are maintained without d
27. ure 40 C Detector UV 254 nm Injection volume 5 0uL Sample 1 Uracil 2 Amitriptyline hydrochloride The chromatogram shown with a dotted line is that of Lot BSIiI03 the previous lot CAPCELL PAK C18 MGI S 5 CAPCELL PAK MG QW Certificate of Analysis Lot BS105 Analysis of Silica gel Specification Result Mean Particle Diameter um 440 4 70 4 63 Particle Distribution 40 90 lt 1 28 1 24 Multi point Nitrogen Sorption Median Pore Diameter nm 9 0 11 0 9 9 Surface Area m g 230 280 258 Pore Volume mL g 0 85 0 99 0 91 1 Quantachrome Autosorb 1 Metal Contents ppm Fe lt 5 0 1 8 Zn lt 5 0 n d Mg lt 5 0 n d Ti lt 5 0 n d Al lt 5 0 3 8 Na lt 5 0 3 0 ICP AES n d not detected Analysis of CAPCELL PAK C18 MGIII S 5 Carbon Content 14 0 15 5 14 4 Chromatographic Results a Hexylbenzene Amylbenzene 1 46 1 50 1 48 k Hexylbenzene 11 3 15 2 12 0 Methyl benzoate Toluene 0 42 0 46 0 42 As Pyridine 0 90 1 50 1 33 As Phenol 0 90 1 20 1 05 As Quinizarin 0 90 1 40 1 06 As Amitriptyline 7 0 90 1 30 1 24 k Amitriptyline 7 13 16 1 3 See Fig 1 on reverse Chromatographic conditions Column 4 6 mm I D x150 mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7L Detector UV 254 nm Temperature 40 C 5 See Fig 2 on reverse ae Chromatographic conditions Column 4 6 mm I D x150 mm Flow rate 1 0 mL m
28. urer is used a separation profile may not be as expected 2 To analyze basic compounds which are protonated under neutral conditions peak shapes became sharper according to the buffer concentration and amount of organic solvent in mobile phase 3 If analysis is performed with a neutral or acidic mobile phase after using a basic or strongly acidic mobile phase retention times and peak shapes of basic compounds may become unstable 2 When using NH2 1 Analysis of carbohydrate Set the mobile phase conditions for a mixed solution of acetonitrile and water As the acetonitrile concentration is higher the carbohydrate retention is greater If methanol or buffer solution is used for the eluate peaks have a tendency to broaden When you try to reproduce the same separation already done with NH2 columns of other manufactures raise the acetonitrile content by 5vol from their conditions Avoid using 100 water in preparing carbohydrate samples Sample solutions of 100 water deteriorate peak shapes by the nature of normal phase chromatography Prepare carbohydrate samples in order to contain acetonitrile for more than 50 2 Analysis of ionic substance Set the mobile phase conditions with a well defined pH value by the use of buffer When optimizing pH begin with a high pH then lower it as needed Once mobile phase of low pH was used the surface of packing material will be changed irreversibly It may take a long time 24 hours
29. x100 mm Flow rate 1 0 mL min Mobile phase Methanol Water 50 50 v v Injection volume 7uL Detector UV 254 nm Temperature 40 C See Fig 2 on reverse i Chromatographic conditions Column 4 6 mm 1 D x 100mm Flow rate 1 0 mL min Mobile phase 20 mmol L KH2PO4 20 mmol L K2HPO Water Methanol 20 80 v v Injection volume 5uL Detector UV 254 nm Temperature 40 C T See Fig 3 on reverse Acid Resistance gt 80 0 83 7 Alkali Resistance gt 70 0 75 7 Chromatographic conditions Column 4 6 mm I D x 100mm Flow rate 1 0 mL min Sample 1000 ppm Benzylalcohol Injection volume 7uL Detector UV 254 nm percentage of k at 20 hr over the initial k Mobile phase Methanol Water TFA 10 89 1 pH 1 0 Temperature 60 C 9 Mobile phase 4 mmol L Sodium Tetraborate Methanol 90 10 pH 10 Temperature 50 C Continued on reverse for further information SHISEIDO CO LTD 7 5 5 GINZA CHUO KU TOKYO 104 0061 JAPAN CAPCELL PAK C18 MGI S 3 Lot BMS1101 min Fig 1 Alkylbenzene 0 3 6 min Fig 2 Pyridine Phenol Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Acetonitrile Water 30 70 v v Flow rate 1 0 mL min Temperature 40 C Detector UV 254 nm Injection volume 7 0uL Sample 1 Pyridine 2 Phenol Chromatographic conditions Column 4 6 mm I D x 100 mm Mobile phase Acetonitrile Water 70 30 v v Flow rate 1 0 mL min Tem
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