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R-Biopharm Rhone Ltd

5

 HPLC Conditions

HPLC Conditions

Derivatisation

KOBRA® CELL at 100 µA setting

Guard Cartridge

Inertsil ODS-3

5 µm, 4 mm x 10 mm or (Hichrom) equivalent

Analytical Column

Inertsil ODS-3V

5 µm, 4.6 mm x 150 mm (Hichrom) or equivalent

Mobile Phase

Water : Methanol (60 : 40 v/v)
Add 119 mg of potassium bromide and 350 µl 4 M Nitric Acid to 1 litre of mobile 

phase

HPLC Pump

To deliver mobile phase

Flow Rate

1.0 ml/minute

Fluorescence 

Detector

Excitation:   362 nm

Emission:     425 nm (B1 and B2)

                     455 nm (G1 and G2)

Column Heater

Maintain guard and analytical columns at 40 °C

Integrator / Data 

Control System

From preferred supplier

Injector

Autosampler / Reodyne valve

Injection Volume

100 µl

Elution Order

G2, G1, B2, B1.

Summary of Contents for KOBRA CELL

Page 1: ...R Biopharm Rh ne Ltd R Biopharm for the safety of your analysis KOBRA CELL Art No K01 Instruction Manual...

Page 2: ...2 R Biopharm Rhone Ltd...

Page 3: ...nents 7 3 2 Precautions When Assembling 8 3 3 Assembly 8 4 KOBRA CELL Installation 9 4 1 Power Pack Notes 9 4 2 Power Pack Technical Specification 9 4 3 Installation Guide 10 4 4 Installation Instruct...

Page 4: ...ks Observed 18 7 3 Abnormal Peaks Observed 19 7 4 Error Light Observed on Power Pack 20 7 5 Leakage from KOBRA CELL 20 7 6 KOBRA CELL Causing High Pressure on the HPLC 20 7 7 Storage Plug Broken Off o...

Page 5: ...lumn The limitations of this method are the use of a second pump and the difficulty in dissolving the PBPB as well as the hazardous nature of the reagent Post column derivatisation with iodine also ha...

Page 6: ...occurs generating the reactive form of bromine The reaction between the reactive bromine and the aflatoxins must take place before the derivatised aflatoxins enter the fluorescence detector Hence the...

Page 7: ...toxins are very hazardous substances Only laboratories equipped to handle toxic materials and solvents should perform analyses Suitable protective clothing including gloves safety glasses and lab coat...

Page 8: ...9 4 Platinum electrode K05 5 PTFE spacer 0 25 mm K07 6 Ion exchange membrane K02 7 PTFE spacer 0 25 mm K07 8 Spacer grid K10 9 PTFE spacer 0 25 mm K07 10 Stainless steel electrode K04 11 PTFE spacer 0...

Page 9: ...ng screws as this will damage the threads Never use metal nuts or ferrules directly with the KOBRA CELL Use a guillotine tubing cutter to prevent distortion of the diameter of the PEEKTM tubing used w...

Page 10: ...required adapter place on the two pins and slide to connect The power pack can now only be used at 100 A and there is a light to show it has been switched on top right There is also an error light to...

Page 11: ...let Outlet Waste Working Electrode platinum Spacer Membrane Spacer Counter Electrode Stainless Steel Membrane Reichelt Thomapor Anion Capillary connections Valco 1 16 inch do not use metal nuts or fer...

Page 12: ...ightened and PEEKTM tubing 7 Connect the KOBRA CELL outlet to waste using plastic ferrules hand tightened and PEEKTM tubing 8 Switch on the HPLC pump and flush mobile phase through the system for appr...

Page 13: ...CELL power pack before turning off the HPLC pump Always use HPLC grade solvents with minimal benzene content from a quality supplier and maintain fresh supplies of potassium bromide Never flush 100 so...

Page 14: ...hase Water Methanol 60 40 v v Add 119 mg of potassium bromide and 350 l 4 M Nitric Acid to 1 litre of mobile phase HPLC Pump To deliver mobile phase Flow Rate 1 0 ml minute Fluorescence Detector Excit...

Page 15: ...he KOBRA CELL leaving the other inlet outlet positions sealed with storage plugs or ferrules 3 Using the syringe with the pipette tip insert this into the empty inlet outlet position on the KOBRA CELL...

Page 16: ...he HPLC pump and flush the system for at least 30 minutes before switching off the system 6 Manually flush the KOBRA CELL through with distilled water several times using a syringe 7 Store the KOBRA C...

Page 17: ...te plastic housing of the KOBRA CELL by removing the 6 locating screws 2 Carefully separate the top white plastic housing and remove in turn the internal layers of the cell using tweezers Make a note...

Page 18: ...2 000 hours depending on the detector Check there is not a problem with the detector Check the standard has been prepared correctly or the standard has not deteriorated Standards should be prepared f...

Page 19: ...use the centre of the membrane becomes discoloured eventually turning a pale colour The pale colour indicates that the membrane will need replaced Check there is power in the power pack Check that th...

Page 20: ...at is at fault Fig 10 Check the correct mobile phase has been used Check the correct HPLC column has been used Check the internal components of the KOBRA CELL have been inserted in the correct order F...

Page 21: ...be replaced 7 5 Leakage from KOBRA CELL The locating screws on the KOBRA CELL have not been tightened correctly Please refer to 3 2 Precautions When Assembling Check the locating screws on the KOBRA C...

Page 22: ...for difficult samples References from the RBR library Installation and support of the KOBRA CELL Advice on detection parameters Advice on preparation and handling of standards Updates on legislation s...

Page 23: ...23...

Page 24: ...R Biopharm Rh ne Ltd R Biopharm Rh ne Ltd Block 10 Todd Campus West of Scotland Science Park Acre Road Glasgow G20 0XA www r biopharm com R Biopharm for the safety of your analysis 28 06 2017 V19...

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