An Evaluation of the Use of Indirect Absorbance for Level Response Detection in HPLC
Brushwyler, Kevin Ross
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The work done for this SIP project explored the limits of application of the indirect photometric detection scheme to a real system containing only neutral components. A discussion of indirect photometric detection and ion pair chromotography is included. For neutral systems, it was discovered that the proposed partial molar volume theory does not adequately describe the interaction between any given neutral sample and neutral UV probe. The experimental results show that cyclohexane gives the expected negative response at the detector. However, spectinomycin, a real pharmaceutical product, appears to form sample-probe complexes which result in positive absorbance peaks rather than the expected negative peaks. Most of the UV absorbers tested exhibited no significant retention on the column. Furthermore, the addition of the UV probe to the mobile phase was found to significantly alter sample retention and resulted in odd column behavior such as steady pressure increases and very long equilibration times.If you are not a current K College student, faculty, or staff member, email firstname.lastname@example.org to request access to this SIP.