Spectroscopic methods of analysis

Colorimetry, in which a sample absorbs visible light, is one example of a spectroscopic method of analysis. At the end of the nineteenth century, spectroscopy was limited to the absorption, emission, and scattering of visible, ultraviolet, and infrared electromagnetic radiation. During the twentieth century, spectroscopy has been extended to include other forms of electromagnetic radiation (photon spectroscopy), such as X-rays, microwaves, and radio waves, as well as energetic particles (particle spectroscopy), such as electrons and ions.  

The focus of this chapter is photon spectroscopy, using ultraviolet, visible, and infrared radiation. Because these techniques use a common set of optical devices for dispersing and focusing the radiation, they often are identified as optical spectroscopies. For convenience we will usually use the simpler term spectroscopy in place of photon spectroscopy or optical spectroscopy however, it should be understood that we are considering only a limited part of a much broader area of analytical methods. Before we examine specific spectroscopic methods, however, we first review the properties of electromagnetic radiation. 

Galego and Arroyo [14] described a simultaneous spectrophotometric determination of OTC, hydrocortisone, and nystatin in the pharmaceutical preparations by using ratio spectrum-zero crossing derivate method. The calculation was performed by using multivariate methods such as partial least squares (PLS)-l, PLS-2, and principal component regression (PCR). This method can be used to resolve accurately overlapped absorption spectra of those mixtures. 

Samola and Urleb [15] reported qualitative and quantitative analysis of OTC using near-infrared (NIR) spectroscopy. Multivariate calibration was performed on NIR spectral data using principle component analysis (PCA), PLS-1, and PCR. 

Fernandez-Gonzales et al. [16] described a method for determination of OTC in medicated premixes and feeds by second-derivative synchronous spectrofluorome-try. The assay based on the reaction of oxytetracycline with divalent metal ion (Ca2+) at pH 6-10 to form a yellow complex that can be analyzed by synchronous spectrofluorometry (AX = 115 nm). Rao et al. [17] described a spectrophotometric method for the determination of OTC in pharmaceutical formulations based on the color reaction with uranium, which was detected at 413 nm. 

In recent years, the chemiluminescence method has been developed for the determi- 

A high performance capillary electrophoresis (HPCE) was described for the separation and simultaneous determination of OTC, TC, CTC, DC, and chloramphenicol in honey. The use of buffer pH 3.2 containing 0.02 mol/L Na2HP04 and 0.01 mol/L citric acid with addition of 4% (v/v) A-methylmorpholine and 12% (v/v) acetonitrile demonstrated a good separation of these five antibiotics within 20 min. The proposed method gave detection limit (signal to noise ratio 5) of 20 pg/L for OTC [26], 

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