Biomedical applications of derivative spectroscopy

Fell, Anthony F.

doi:10.1016/0165-9936(83)85010-9

Cited by 81 publications

(28 citation statements)

References 33 publications

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“…Very little additional laboratory time is required for the spectrofluorometer to display the firstderivative spectra. Undergraduate students are exposed to the general method of derivative spectroscopy, which is an important, often-used analytical technique (15)(16)(17)(18)(19)(20) for eliminating sample matrix and background fluorescence effects and for treating overlapped spectral bands. Moreover, students have the opportunity to actually perform the statistical analysis involved with comparing different analytical methods.…”

Section: Discussion Of Resultsmentioning

confidence: 99%

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Comparison of Analytical Methods: Direct Emission versus First-Derivative Fluorometric Methods for Quinine Determination in Tonic Waters

et al. 1999

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Fluorescence spectroscopy is an extremely versatile, sensitive experimental technique used in identification and quantification of many environmentally important compounds such as polycyclic aromatic hydrocarbons, polycyclic aromatic nitrogen heterocycles, and polycyclic aromatic sulfur heterocycles. Through judicious selection of excitation and emission wavelengths, a single desired fluorophore can often be analyzed in complex unknown mixtures containing several absorbing and fluorescing species.Many laboratory experiments appearing in this Journal (1-9) and standard laboratory manuals (e.g., ref 10) have involved determination of analyte concentrations by fluorometric methods. Published methods assume that the observed emission intensity, F, isdirectly proportional to the molar concentration of the analyte. The proportionality constant, K′, depends upon the quantum efficiency (quantum yield) of the fluorescence process, the response of the photodetector at the emission wavelength, and the molar extinction coefficient, which remain constant during any given chemical analysis at fixed excitation and emission wavelengths. Analyte concentrations are determined from a working-curve plot of the measured fluorescence intensity versus the known molar concentrations of the standard solutions. The aforementioned experimental methods introduce students to fluorescence instrumentation. However, the data analysis will appear rather trivial if UV-vis spectrophotometric, flame emission, or AA analysis has already been performed. Most instrumental analysis textbooks (11-14) discuss absorption spectroscopy and applications of the Beer-Lambert law one or two chapters before presenting fluorescence and phosphorescence.We have found it possible to modernize our existing fluorometric laboratory experiment involving the determination of quinine in tonic waters by statistically comparing values determined from direct emission and first-derivative fluorometric methods. Recent review articles (15-20), written in several different languages, have cited numerous examples of the application of derivative spectroscopy to the analysis of food, clinical, pharmaceutical, biomedical, and environmental samples. For the most part, published applications utilize either the first or second derivative. Third and higher-order derivatives have been successfully used in select occasions. The first-derivative spectrofluorometric method is relatively straightforward and will be discussed in terms of an unknown tonic water sample containing quinine. The measured emission intensity is given by eq 1. Differentiation of the solution fluorescence emission with respect to the emission wavelength, λ em , yields the following mathematical expression:For solutions that contain only a single fluorophore, the first derivative corresponds to the gradient dF/d λ em of the fluorescence emission envelope and for each well-resolved band features only a maximum and trough. The vertical distance is the amplitude, which is directly proportional to the analyte concentrati...

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Section: Discussion Of Resultsmentioning

confidence: 99%

“…Recent review articles (15)(16)(17)(18)(19)(20), written in several different languages, have cited numerous examples of the application of derivative spectroscopy to the analysis of food, clinical, pharmaceutical, biomedical, and environmental samples. For the most part, published applications utilize either the first or second derivative.…”

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confidence: 99%

Comparison of Analytical Methods: Direct Emission versus First-Derivative Fluorometric Methods for Quinine Determination in Tonic Waters

et al. 1999

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“…5 B, C). These minima at 280 nm ( f 2 nm) and 290 nm ( f. 2 nm) are typical of tyrosine and tryptophan residues respectively [14]. A detailed spectral analysis of a number of synthetic peptides (data not given), containing various combinations of aromatic amino acids, indicates that the tryptophan extremum often masks the tyrosine extremum when the tryptophan-totyrosine ratio is about 1 : 1.…”

Section: Spectral Analysis Of Purqied Tryptic Peptidesmentioning

confidence: 99%

A microbore high-performance liquid chromatography strategy for the purification of polypeptides for gas-phase sequence analysis. Structural studies on the murine transferrin receptor

et al. 1985

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We describe herein the use of reversed-phase high-performance liquid chromatography coupled with the novel application of short (10 cm or less) microbore columns (2 mm internal diameter) to fractionate and purify a number of tryptic peptides generated from approximately 200 pmol purified murine transferrin receptor. The use of reversed-phase microbore columns permits the recovery of submicrogram amounts of purified polypeptides in high yield (greater than 90%) in small eluent volumes (20-60 pl). In this manner, purified polypeptides can be loaded directly onto the gas-phase sequencer without further manipulation. This procedure avoids sample loss, which frequently occurs with other forms of concentration (e. g. lyophilization, evaporation). The application of second-order-derivative ultraviolet spectroscopy, using a diode array detector, for the analysis of aromatic aminoacid-containing peptides in complex tryptic digests is described. N-terminal amino acid sequence analyses were performed on six tryptic peptides, yielding 105 unique assignments; this corresponds to approximately 14% of the molecule. A comparison of this amino acid sequence information with the primary structure of human transferrin receptor deduced from the mRNA sequence [Nature (Lond.) 311, 675-678 (1984); Cell 39,267 -274 (1984)l reveals, with the exception of one tryptic peptide, a very close sequence homology between the murine and human transferrin receptors.The receptor for the iron-binding protein transferrin is an integral membrane glycoprotein located on the surface of all proliferating mammalian cells [l]. Recent evidence indicates that the transferrin receptor is a dimer composed of two similar or identical polypeptide chains ( M , 95 000) joined by disulfide bonds [2 -41. The recent purification to homogeneity [5] of small quantities (200 pmol) of the murine transferrin receptor from NS-1 myeloma cells has led us to undertake a partial amino acid sequence analysis of this protein. The partial sequence information obtained in this manner has been used to construct the appropriate oligonucleotide probes, thereby permitting the primary structure determination of the murine transferrin receptor using recombinant DNA procedures [6].Direct amino acid sequence analysis of high-M, proteins often yields little meaningful sequence information. As this was the case with the murine transferrin receptor, we decided to fragment the parent molecule into smaller peptides (e. g. tryptic peptides), which are more suitable for sequence analysis. Crucial to the success of this strategy is the ability to purify to homogeneity submicrogram amounts of some of the resultant peptides, and to recover them in volumes suitable for gas-phase sequence analysis. To this end we demonstrate

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“…This increased selectivity results because bands that are overlapped in zeroorder absorption spectra can be separated in the derivative mode. Theoretical and practical studies done by O'Haver and Green 10,11 and Fell and co-workers 12,13 showed that this technique can lead to a faster and more accurate determination of multi-component mixtures that previously would have required time-consuming separation techniques. Rhodium and iridium were preconcentrated from a fairly large volume of their aqueous solutions using 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) and tetraphenylborate onto microcrystalline naphthalene in the pH ranges 5.0 -6.5 and 3.5 -5.5, respectively.…”

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confidence: 99%

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confidence: 99%

Simultaneous Determination of Rhodium and Iridium Using Derivative Spectrophotometry after Preconcentration of Their 2-(5-Bromo-2-pyridylazo)-5-diethylaminophenol Chelates onto Microcrystalline Naphthalene

Pancras

Puri

1999

ANAL. SCI.

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The economy and efficiency of the analytical procedure for the separation and determination of precious metals, such as rhodium and iridium in geological samples and related materials, are still a challenge to the analyst. Because of their low concentration levels in natural sources, a necessary first step in any analytical procedure needs to be the preconcentration of these elements. The most common and well-documented concentration-cum-separation technique is liquid-liquid extraction. However, this technique is tedious, time consuming (extraction involves several steps and equilibrium distribution between the two phases takes longer time in certain cases) and often requires a large volume of high purity toxic solvents. Column methods using various adsorbents are effective for the preconcentration of metal ions. But their main disadvantages are the lengthy method to prepare the column materials and other stringent conditions. The sorption and desorption of metal ions are carried out at low flow rates, which make these methods time consuming.1,2 To overcome these drawbacks, Satake and co-workers 3,4 developed a method called "Solid-liquid separation after the adsorption of metal-chelates onto microcrystalline naphthalene". This method is very simple, economic and less time-consuming.Only a few photometric reagents have been used for the trace determination of rhodium and iridium in solutions. The similarities in chemical properties of rhodium and iridium make it difficult to eliminate the interference between them in zero-order (normal) spectrophotometry. Tertipis and Beamish 5 separated and determined rhodium and iridium using tin(II) bromide. Stokely and Jocobs 6 proposed a simultaneous zeroorder spectrophotometric determination using 1-(2-pyridylazo)-2-naphthol. Zhenya 7 reported a simultaneous dual-wavelength spectrophotometric determination using tin(II) bromide and 2-mercaptobenzothiazole. Shkil 8 separated rhodium from iridium in acid medium using 2-mercaptobenzothiazole. Wilson and Jacobs 9 separated relatively a large amount of iridium from rhodium by extracting iridium into tributyl phosphate. All of these methods utilized time-consuming solvent extraction for separation. Moreover, they are not sufficiently sensitive and selective. Derivative spectrophotometry has the advantage of higher selectivity than zero-order spectrophotometry. This increased selectivity results because bands that are overlapped in zeroorder absorption spectra can be separated in the derivative mode. Theoretical and practical studies done by O'Haver and Green 10,11 and Fell and co-workers 12,13 showed that this technique can lead to a faster and more accurate determination of multi-component mixtures that previously would have required time-consuming separation techniques. Rhodium and iridium were preconcentrated from a fairly large volume of their aqueous solutions using 2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (5-Br-PADAP) and tetraphenylborate onto microcrystalline naphthalene in the pH ranges 5.0 -6.5 and 3.5 -5.5, re...

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Biomedical applications of derivative spectroscopy

Cited by 81 publications

References 33 publications

Comparison of Analytical Methods: Direct Emission versus First-Derivative Fluorometric Methods for Quinine Determination in Tonic Waters

Comparison of Analytical Methods: Direct Emission versus First-Derivative Fluorometric Methods for Quinine Determination in Tonic Waters

A microbore high-performance liquid chromatography strategy for the purification of polypeptides for gas-phase sequence analysis. Structural studies on the murine transferrin receptor

Simultaneous Determination of Rhodium and Iridium Using Derivative Spectrophotometry after Preconcentration of Their 2-(5-Bromo-2-pyridylazo)-5-diethylaminophenol Chelates onto Microcrystalline Naphthalene

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