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LABORATORY PROCEDURES FOR PLANT CELL LINES

REFERENCE NO.: PC/1998/3/1.3


TITLE: STANDARD HPLC PROCEDURE FOR CULTURE EXTRACTS


INTRODUCTION

After extraction of the plant cell cultures, the resulting extracts are analysed by a standard HPLC procedure.

PROCEDURE

1. Assure that all single electrical devices of the analytical equipment are set to position: on.

(The following equipment is used: solvent delivery system: 2 pumps contraMetric 3000, LDC Analytical, 1990; detector: SM4000, LDC Analytical, 1990; autosampler: Marathon, Spark Holland, 1990; Dynamic + Static HPLC Mixing Chamber, Spark Holland, 1990; pump control and data analysis system: Thermochrom Vers. IV, LDC Analytical, 1990.)

2. Prepare the necessary volumes of solvent A and solvent B according to the standard procedure (PC/1998/3/1.5) and refill the solvent reservoirs of the HPLC pumps.

3. Place the vials to be analysed in the autosampler and program the autosampler according to the manual.

4. Make sure that the parameters of the UV detection unit are adjusted to: wavelength = 280 nm, range = 0.01.

5. Program the sequence for the analysis of this set of samples according to the manual.

5.1. Use Method 6 for equilibrating the column before the analytical runs (PC/1998/3/1.6)

5.2. Use Method 1 for the standard analysis of samples (PC/1998/3/1.6)

5.3. Use Method 7 for equilibrating the column after the analytical runs (PC/1998/3./1.6)

6. Start the programmed analytical sequence at the computer.

7. At the end of method 6 start the analytical sequence at the autosampler.

8. When the programmed analytical sequence of samples is finished, print out the single profiles and copy the single run profiles into the corresponding storage files

(c:\maa\lc for collection number l; c:\maa\nac for collection number na; c:\maa\zac for collection number za; c:\maa\zcc for collection number zc).

9. Set all single parts of the HPLC device to position: off


Guidelines prepared for CABRI by DSMZ, 20 Jan. 1998; updated August 1999
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Copyright CABRI, 1998 

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