Hewlett-Packard fluorescence detector

Figure 3.16—Flow cell of a fiuorimetric detector (reproduced by permission of Hewlett-Packard Inc.). One example of a reagent that is used to form fluorescent derivatives with compounds containing primary amines. Reaction of OPA in the presence of monothioglycol.

The HPLC was a Du Pont 8800 quaternary solvent system with a Hewlett-Packard (HP) 1040A photodiode array detector (5-8). Flexible disks were used for data storage, and postrun data evaluation was performed by the detector s computer (HP 85). Samples were injected with a loop injector 10 p,L was used for analytical scale, and 200 jlL was used for preparative scale. Spectra were run on a spectrophotometer (Perkin-Elmer Lambda 3) for static absorbance and on a spectrofluo-rometer (Perkin-Elmer MPF-66) for fluorescence. Field-ionization mass spectra were obtained at a resolution of 45,000 (corresponding to a 0.01-daltons error for a mass of 450). 

Hi h Pressure Liquid Chromatography. Size exclusion chromatography was performed isocratically in 0.2 M ammonium acetate (flow-1.5 ml/min) with a Water s Model 840 HPLC with a Toyo-Soda G2000SWXL column (30 cm x 7.8 mm). Eluting components were monitored in the UV at 214 nm and by fluorescence (Ex - 230 nm, Em - 300 nm) and collected in 4 minute fractions for bioassay. Reverse phase chromatography was conducted with a Hewlett-Packard Model 1090 HPLC equipped with a Vydac C-18 column using a linear gradient over one hr from 10% to 50% acetonitrile in 0.1% TFA. Sample was monitored with a diode array detector from 190 to 350 nm in the UV. 

Hewlett-Packard HP 1100 Quaternary pump w/degasser, thermostatted column compartment, w/fluorescence and refractive index detectors... 

The chromatographie system eonsisted of a quaternary pump, an automatic sample injector (Hewlett-Packard 1050 Series, Palo Alto, CA, USA) and a programmable fluorimetric detector (Hewlett-Packard, 1046 Series). The detector was linked to a data system (Hewlett-Paekard, HPLC ChemStation) for data acquisition and calculation. According to the results of preliminary experiments obtained by injecting directly into the analytical column standard solutions of PAHs, the deteetor was programmed to measure the fluorescence intensity at the exeitation/emission wavelengths pairs listed in Table 1. 

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