Liquid chromatography vitamins analysis

Siluk D, Oliveira RV, Esther-Rodriguez-Rosas M et al. A validated liquid chromatography method for the simultaneous determination of vitamins A and E in human plasma. Journal of Pharmaceutical and Biomedical Analysis, 44, 1001-1007, 2007. 

Currently, high-performance liquid chromatography (HPLC) methods have been widely used in the analysis of tocopherols and tocotrienols in food and nutrition areas. Each form of tocopherol and tocotrienol can be separated and quantified individually using HPLC with either a UV or fluorescence detector. The interferences are largely reduced after separation by HPLC. Therefore, the sensitivity and specificity of HPLC methods are much higher than those obtained with the colorimetric, polarimetric, and GC methods. Also, sample preparation in the HPLC methods is simpler and more efficiently duplicated than in the older methods. Many HPLC methods for the quantification of tocopherols and tocotrienols in various foods and biological samples have been reported. Method number 992.03 of the AOAC International Official Methods of Analysis provides an HPLC method to determine vitamin E in milk-based infant formula. It could probably be said that HPLC methods have become dominant in the analysis of tocopherols and tocotrienols. Therefore, the analytical protocols for tocopherols and tocotrienols in this unit are focused on HPLC methods. Normal and reversed-phase HPLC methods are discussed in the separation and quantification of tocopherols and tocotrienols (see Basic Protocol). Sample... 

Sanchez-Perez, A., Delgado-Zamarreno, M.M., Bustamante-Rangel, M., and Hernandez-Men-dez, J. 2000. Automated analysis of vitamin E isomers in vegetable oils by continuous membrane extraction and liquid chromatography-electrochemical detection. J. Chromatogr. A 881 229-241. 

JN Thompson. Trace analysis of vitamins by liquid chromatography. In JF Lawrence, ed. Trace Analysis. Vol 2. New York Academic Press, 1982, pp 1-67. 

DB Dennison, JR Kirk. Quantitative analysis of vitamin A in cereal products by high-speed liquid chromatography. J Food Sci 42 1376-1379, 1977. 

F Zonta, B Stancher. Quantitative analysis of phylloquinone (vitamin K,) in soy bean oils by high-performance liquid chromatography. J Chromat 329 257-263, 1985. 

DA Sampson, LA Eoff, XL Yan, K Lorenz. Analysis of free and glycosylated vitamin B6 in wheat by high-performance liquid chromatography. Cereal Chem 72 217-221, 1995. 

M Careri, R Cilloni, MT Lugari, P Manini. Analysis of water-soluble vitamins by high-performance liquid chromatography-particle beam-mass spectrometry. Anal Commun 33 159-162, 1996. 

Figure 1 is the ultraviolet spectrum of a 10 mcg/ml solution of vitamin D3 in methanol. The spectrum was obtained using a Cary Model 219 recording spectrophotometer (Varian Instrument Co., Palo Alto, CA). Vitamin D3 and related compounds have a characteristic UV absorption maximum at 265 nm and a minimum at 228 nm. The extinction coefficient at 265 nm is about 17,500 and 15,000 at 254 nm. An index of purity of vitamin D3 is a value of 1.8 for the ratio of the absorbance at 265 to that at 228 nm. The high absorbance at 254 nm enables one to use the most common and sensitive spectrophotometric detector used in high performance liquid chromatography (HPLC) for the analysis of vitamin D3 in multivitamin preparations, fortified milk, other food products, animal feed additives etc. 

Chavez-Servin, J.L. Castellote, A.I. Lopez-Sabater, M.C. 2006. Simultaneous analysis of vitamins A and E in infant milk-based formulae by normal-phase high-performance liquid chromatography-diode array detection using a short narrow-bore column. J. Chromatogr. A. 1122 138-143. 

The quantitative analysis of the fat-soluble vitamins (A, E, D and K) and their esters by reversed-phase partition in water/alcohol solvents on Zipax columns has been reported [255]. The applicability of gas and high pressure liquid chromatography of vitamin A was discussed by Vecchi, Vesely and Oesterhelt [256] who concluded that HPLC was superior in this application. 

Preferably, high pressure liquid chromatography (hplc) is used to separate the active pre- and cis-isomers of vitamin D3 from other isomers and allows theic analysis by comparison with the chromatograph of a sample of pure reference j-vitamin D3, which is equilibrated to a mixture of pre- and cis-isomers (82,84,85). This method is more sensitive and provides information on isomer distribution as well as the active pre- and cis-isomer content of a vitamin D sample. It is applicable to most forms of vitamin D, including the more dilute formulations, ie, multivitamin preparations containing at least 1 lU/g (AOAC Methods 979.24 980.26 981.17 982.29 985.27) (82). The practical problem of isolation of the vitamin material from interfering and extraneous components is the limiting factor in the assay of low level formulations. 

Delgado Zamarreno MM, Sanchez Perez A, Bustamante Rangel M, and Hernandez Mendez J. Automated analysis for vitamin E in butter by coupling sample treatment-continuous membrane extraction—liquid chromatography with electrochemical detection. Anal. Chim. Acta 1999 386 99-106. 

Kramer, J.K.G., Fouchard, R.C., and Kallury, K.M.R. 1999. Determination of Vitamin E Forms in Tissues and Diets by High-Performance Liquid Chromatography Using Normal-Phase Diol Column, in Analysis of Vitamin E Forms. New York, NY Academic Press Ltd., pp. 318-329. 

A number of very good reviews on food analysis can be found in the literature [7-12]. Table 3 presents a very limited representation of the kind of work involved in a food laboratory. All basic constituents of foodstuffs - proteins, lipids, carbohydrates and vitamins - are amenable to liquid chromatography. Various types of columns and detectors used for those analysis demonstrate the versatility of the technique. Almost any type of food matrix can be extracted in order to identify and quantitate trace amounts of analytes. 

EXPERIMEIMT 33 QUALITATIVE AND QUANTITATIVE ANALYSIS OF FRUIT JUICES FOR VITAMIN C USING HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY ... 

Williams MV, Lee SH, Blair lA. Liquid chromatography/mass spectrometry analysis of bifunctional electrophiles and DNA adducts from vitamin C mediated decomposition of 15-hydroperoxyeicosatetraenoic acid. Rapid Commun Mass Spectrom 2005 19 849-858. 

Fluorescence spectroscopy plays an important function in modern food analysis as can be seen from its wide use in the determination of numerous food components, contaminants, additives, and adulterants. This technique has made available very sensitive and selective methods that satisfy the requirements of food analysis, which are usually very complex, taking into account the large number of species to be determined, frequently at very low concentrations, and the wide variety of foodstuffs available. Initially, the use of fluorescence spectroscopy in food analysis was limited to the determination of species with intrinsic fluorescence (e.g., vitamins, aflatoxins, and some polycyclic aromatic hydrocarbons (PAHs)), but now it is widely applied to nonfluorescent species, using several physicochemical means such as chemical or photochemical derivatization reactions. Numerous techniques involve fluorescence detection in liquid chromatography (LC), frequently using pre- or postcolumn derivatization. In addition to conventional fluorime-try, which is commonly chosen for this purpose, other fluorimetric techniques such as laser-induced... 

See also Amino Acids. Electrophoresis Proteins. Food and Nutritional Analysis Overview Antioxidants and Presen/atives. Immunoassays, Applications Food. Immunoassays, Techniques Luminescence Immunoassays. Kinetic Methods Principles and Instrumentation. Liquid Chromatography Food Applications. Proteins Foods. Vitamins Overview. 

See also Amperometry. Derivatization of Anaiytes. Food and Nutritional Analysis Meat and Meat Products Dairy Products. Liquid Chromatography Food Applications. Nitrogen. Polarography Inorganic Applications. Quality Assurance Primary Standards. Spectrophotometry Organic Compounds. Sulfur. Vitamins Fat-Soluble Water-Soluble. 

Inductively Coupled Plasma. Carbohydrates Sugars -Spectrophotometric Methods Starch. Food and Nutritional Analysis Oven/iew Antioxidants and Presen/atives Oils and Fats. Ion-Selective Electrodes Food Applications. Liquid Chromatography Amino Acids. Proteins Foods. Quality Assurance Quality Control. Sampling Theory. Vitamins Fat-Soluble Water-Soluble. Water Determination. 

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