Efficiency of Sun<strong>Shell</strong> C18UHPLCColumn: Sun<strong>Shell</strong> C18, 50 x 2.1 mm3Plate(3)=10,3001240 1 2 3 4 5Retention time/minColumn: Sun<strong>Shell</strong> C18, 100 x 2.1 mm31 2Plate(3)=21,3004Column: Sun<strong>Shell</strong> C18, 2.6 µm 50 x 2.1 mmMobile phase: CH 3 CN/H 2 O=60/40Flow rate: 0.3 mL/minPressure: 7 MPaTemperature: 23 ºCSample: 1 = Uracil2 = Toluene3 = Acenaphthene4 = ButylbenzeneColumn: Sun<strong>Shell</strong> C18, 2.6 µm 100 x 2.1 mmMobile phase: CH 3 CN/H 2 O=60/40Flow rate: 0.3 mL/minPressure: 12.5 MPaTemperature: 25 ºC0 2 4 6 8Retention time/minColumn: Sun<strong>Shell</strong> C18,150 x 4.6 mmEfficiency=253,000 plate/m1120 2 4 6 8 10Retention time/minColumn: Sun<strong>Shell</strong> C18, 150 x 4.6 mm32Plate(3)=38,00034Plate(3)=31,0004HPLC80% efficiency tocompare with UHPLCColumn: Sun<strong>Shell</strong> C18, 2.6 µm 150 x 4.6 mmSun<strong>Shell</strong> C18, 2.6 µm 100 x 4.6 mmMobile phase: CH 3 CN/H 2 O=70/30Flow rate: 1.0 mL/minPressure: 14.5MPa(UHPLC),13.5 MPa(HPLC) for 150 mm9.5MPa(HPLC) for 100 mmTemperature: 25 ºCSample: 1 = Uracil2 = Toluene3 = Acenaphthene4 = Butylbenzene1Column: Sun<strong>Shell</strong> C18, 100 x 4.6 mm23Plate(3)=20,00040 2 4 6 8 10Retention time/min0 2 4 6 8 10Retention time/minLaChrom ELITEThe same efficiency as5 µm, 250 x 4.6 mmSaving 60% foranalytical time andconsumption of solventWebsite : www.chromtech.net.au E-mail : info@chromtech.net.au TelNo : 03 9762 2034 . . . in AUSTRALIA
Examples of transfer from a conventional 5 µm column to Sun<strong>Shell</strong> columnIsocratic separationHPLC12311/3 of analysis time0 2 4 6 8 10 12 14 16 18 20 22 24Retention time/minN(4)=20,2871Sun<strong>Shell</strong> C18, 2.6 µm 100 x 4.6 mm452233ACE C18, 5 µm 250 x 4.6 mm0 1 2 3 4 5 6Retention time/min44N(4)=19,3135N(4)=20,2875Column:ACE C18, 5 µm 250 x 4.6 mmSun<strong>Shell</strong> C18, 2.6 µm 100 x 4.6 mmMobile phase:CH 3 CN/20mM Phosphoric acid = 45/55Flow rate: 1.0 mL/min,1.8 mL/min at the lowest chromatogramTemperature: 25 ºCPressure: 9.5 MPa for ACE C18 5 µm13.4 MPa for Sun<strong>Shell</strong> C18 2.6 µmDetection: UV@230 nmSample: 1 = Benzydamine2 = Ketoprofen3 = Naproxen4 = Indomethacin5 = IbuprofenUHPLC1231.0 mL/min4N(4)=24,1245HPLC: Hitachi LaChrom ELITE (using 0.25 mm i.d. tubing)UHPLC: Jasco X-LC123451.8 mL/minN(4)=22,424Analysis time decreases to 1/6 to comparewith a 5 µm column sized 250 x 4.6 mm.0 1 2 3 4 5 6Retention time/minGradient separationHPLC140 2 4 6 8 10 12 14 16 18 20 22 24Retention time/min12Sunniest C18, 5 µm 150 x 4.6 mm340 1 2 3 4 5 6Retention time/min152Sun<strong>Shell</strong> C18, 2.6 µm 75 x 4.6 mm6728 93341/2 of Analysis time5656778 98 97 8 9Column:Sunniest C18, 5 µm 150 x 4.6 mmSun<strong>Shell</strong> C18, 2.6 µm 75 x 4.6 mmMobile phase:A) 0.1% Phosphoric acidB) CH 3 CNGradient program for Sunniest C18for Sun<strong>Shell</strong> C18Time 0 min 15 min 20 min%B 2% 25% 25%Time 0 min 7.5 min 10 min%B 2% 25% 25%Flow rate: 1.0 mL/min,Temperature: 25 ºCDetection: UV@250 nmSample: Oolong tea1 = Gallocatechin, 2 = Epigallocatechin,3 = Catechin, 4 = Caffeine, 5 = Epicatechin,6 = Epigallocatechin gallate, 7 = Gallocatechingallate, 8 = Epicatechin gallate, 9 = Catechin gallateUHPLC120 1 2 3 4 5 6Retention time/min345678 9HPLC: Hitachi LaChrom ELITE (using 0.25 mm i.d. tubing)UHPLC: Jasco X-LCThere are difference of system time lagbetween HPLC and UHPLC. UHPLC has muchless system time lag than HPLC because of highpressure gradient system for UHPLC and lowpressure gradient system for HPLC.Website : www.chromtech.net.au E-mail : info@chromtech.net.au TelNo : 03 9762 2034 . . . in AUSTRALIA