Introduction

Fluorescence detection has been widely used as one of a practical detection method in HPLC analysis. It enables the selective detection of fluorescent substances, with sensitivity that can be up to 1000 times greater than with UV detection.

Fluorescence detection can also be applied to non-fluorescent substances with the use of derivatization, with many derivatization agents becoming commercially available in recent years. Therefore, this technique provides the advantages of highly sensitivity and selectivity to many different classes of compound, and broadens the range of applications in many industries. However, fluorescence detection has not been used in SFC analysis due to the difficulty of developing a suitable high pressure flow cell.

We developed a fluorescence detector with a flow cell specifically constructed for operations at the high pressures required for SFC. This fluorescence detector is included in our newer SFC system (SFC-4000 series). In this presentation, we will introduce an application for the analysis of several compounds using SFC separation with fluorescence detection.

Experimental

Apparatus

Figure 1 shows the appearance of JASCO SFC system equipped with UV/Vis (UV) and fluorescence (FL) detector used in this experiment. Figure 2 also shows the schematic diagram of this system. This system enables to perform the method scouting analysis on mobile phases and columns.

Figure 3 shows the structures and differences between the conventional and novel FL flow cell.  We developed newly quartz flow cell design and structure against high pressure (up to 20 MPa). The outer surface of this flow cell are coated with aluminum, it can realize high collection efficiency of fluorescence by functioning as a reflector.

Results

SFC/UV vs. SFC/FL

SFC/FL Applications

Conclusion

– We developed a fluorescence detector with a flow cell specifically constructed for operation at the high pressures required for SFC
– SFC/FL provides highly sensitive detection (from fg to pg) for fluorescent substances, compared to UV/Vis detector.
– SFC equipped with the fluorescence detector will open the door to new applications fields.

References

Poster session at SFC 2017 at Rockville, Maryland USA
Satoe Iijima¹, Akitaka Terada¹, DJ Tognarelli², John Burchell², Takeshi  Kanomata¹, Masao Bounoshita¹, Yasuyo Sato¹, Miki Kuwajima¹
1JASCO Corporation, 2967-5 Ishikawa-machi, Hachioji, Tokyo 192-8537
2JASCO Incorporated, 28600 Mary’s Court, Easton, MD 21601

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3.O. A. Adegoke, Afr.J. Pure Appl.Chem., 2012, 6 (14), 129-140.
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