Carotenoid food analysis

Mass spectrometry has become an essential analytical tool for a wide variety of biomedical applications such as food chemistry and food analysis. Mass spectrometry is highly sensitive, fast, and selective. By combining mass spectrometry with HPLC, GC, or an additional stage of mass spectrometry (MS/MS), the selectivity increases considerably. As a result, mass spectrometry may be used for quantitative as well as qualitative analyses. In this manual, mass spectrometry is mentioned frequendy, and extensive discussions of mass spectrometry appear, for example, in units describing the analyses of carotenoids (unitfia) and chlorophylls (unit F4.5). In particular, these units include examples of LC/MS and MS/MS and the use of various ionization methods. 

Due to the health-promoting properties associated with the consumption of carotenoids, the analysis of these compounds in plasma, serum, and tissues has become important. The same precautions used when extracting carotenoids from foods must also be employed here to minimize isomerization, oxidation, and degradation. 

Of course, the term proven efficacy is central to any resource investment in this area. Basic information on time and dose responses in humans to complex foods rich in carotenoids (and other phytochemicals) is pitifully small. Much of our information is based upon inadequate databases derived from chemical analysis, in vitro models that have not been properly evaluated or validated, and short-term, high-dose human studies. Future research progress requires much more rigorous debate on the experimental systems employed... 

HART J D and SCOTT K J (1995) Development and evaluation of an HPLC method for the analysis of carotenoids in foods and the measurement of carotenoid content of vegetables and fruits commonly consumed in the UK. Food Chem. 54(1) 101-111. 

Rodriguez-Amaya, D.B. and Kimura, M., Harvest Plus Handbook for Carotenoid Analysis Harvest Plus Technical Monograph 2, Washington, International Food Policy Research Institute and International Center for Tropical Agriculture, 2004. 

Rodriguez- Amaya, DL., A Guide To Carotenoid Analysis in Foods, International Life Sciences Instimte Press, Washington, D.C., 2001. 

Insights into the mechanisms of carotenoid degradation can be followed in model systems that are more easily controlled than foods and the formation of initial, intermediate, and final products can also be more easily monitored. However, extrapolation to foods must be done with caution because simple model systems may not reflect the nature and complexity of a multicomponent food matrix and the interactions that can occur. In addition, even in model systems, one must keep in mind that carotenoid analysis and identification are not easy tasks. 

Schoefs, B., Chlorophyll and carotenoid analysis in food products. A practical case-by-case view. Trends Anal. Ghent., 22, 335, 2003. 

On the other hand, quantitative extraction requires complete and exhaustive extraction and no material can be lost. To assure complete extraction when a food is analyzed for the first time in a laboratory, it is useful to carry out two or three extractions, pool the solvents, and keep separate the next extracts to verify the presence of carotenoids. Usually four to six extractions are enough to remove the carotenoids completely from a sample. The extraction can be carried out in a blender, vortex, or with a mortar and pestle. Accelerated solvent extraction (ASE), an important extraction technique in residue analysis, currently attracts interest due to its short duration, low level of solvent use, and high extraction yield. The average recoveries for all carotenoids with the exception of norbixin ranged from 88.7 to 103.3% using manual extraction and from 91.0 to 99.6% by ASE (70 bar and temperature of 40°C) both extractions were carried out with a mixture of MeOH, EtOAc, and petroleum ether (1 1 1). ... 

No expensive equipment is required for OCC however, the separation efhciency depends on the analyst s experience since a new column has to be packed for each analysis. In addition, depending on the packing type (powder or slurry), stationary phase, and purpose of the separation, the separation can take from 30 min to 4 hr. The AOAC official method for the determination of carotenoids still uses OCC." Separation of carotenoids from many foods was developed on a column packed with a mixture of MgO and HyfloSupercel (or celite or diatomaceous earth) at 1 1... 

Mercadante A.Z., Rodriguez-Amaya, D.B., and Britton, G., HPLC and mass spec-trometric analysis of carotenoids from mango, J. Agric. Food Chem., 45, 120, 1997. 

Scott, K. et al.. Interlaboratory studies of HPLC procedures for the analysis of carotenoids in foods. Food Chem., 57, 85, 1996. 

Carmen Socaciu was bom in Cluj-Napoca, Romania and earned a BSc in chemistry in 1976, an MSc in 1977, and a PhD in 1986 from the University Babes-Bolyai in Cluj-Napoca, an important academic centre located in the Transylvania region. Dr. Socaciu worked as a researcher in medical and cellular biochemistry for more than 10 years, and became a lecturer in 1990 and full professor in 1998 in the Department of Chemistry and Biochemistry of the University of Agricultural Sciences and Veterinary Medicine (USAMV) in Cluj-Napoca. She extended her academic background in pure chemistry (synthesis and instrumental analysis) to the life sciences (agrifood chemistry and cellular biochemistry). Her fields of competence are directed especially toward natural bioactive phytochemicals (carotenoids, phenolics, flavonoids), looking to advanced methods of extraction and analysis and to their in vitro actions on cellular metabolism, their effects as functional food ingredients, and their impacts on health. 

Partali, V., Liaaen-Jensen, S., Slagsvold, T., and Lifjeld, J. T. 1987. Carotenoids in food chain studies. II. The food chain of Pams spp. monitored by carotenoid analysis. Comp. Biochem. Physiol. B 87 885-888. 

The foliage of the food plants was ground and the pigments were extracted into warm methanol and saponified in 4% sodium hydroxide. The carotenoids were extracted into dichloromethane, dried, and redissolved in ethanol prior to an analysis by HPLC. 

The HPLC analysis of milkweed, the food-plant source for Monarch butterflies, demonstrates that it contains a complex mixture of carotenoids including lutein, several other xanthophylls, xanthophyll epoxides, and (3-carotene, Figure 25.3b. There is a component in the leaf extract that is observed to elute near 8min, which has a typical carotenoid spectrum but is not identical to that of the lutein metabolite observed at near the same retention time in the extracts from larval tissue. 

Mercadante AZ, Rodriguez-Amaya DB and Britton G. 1997. HPLC and mass spectrometric analysis of carotenoids from mango. J Agric Food Chem 45 120—123. 

Ben-Amotz A and Fishier R. 1998. Analysis of carotenoids with emphasis on 9-cis-fS-carotene in vegetables and fruits commonly consumed in Israel. Food Chem 62 515-520. 

Rodriguez-Amaya D. 2001. A Guide to Carotenoid Analysis in Foods. Washington, DC ILSI Press, International Life Science Institute. 

A.M. Pupin, M.J. Dennis and M.C.F. Toledo, HPLC analysis of carotenoids in orange juice. Food Chem. 64 (1999) 269-275. 

M. Baranska, H. Schulz, R. Baranski, T. Nothnagel and L.P. Christensen, In situ simultaneous analysis of polyacetylenes, carotenoids and polysaccharides in carrot roots, J. Agric. Food Chem., 53, 6565-6571 (2005). 

Recently we published data that even in countries with excellent food sources and availability, insufficient vitamin A supply will occur (Schulz et ah, 2007). The aim of this trial was to analyze vitamin A and p-carotene status and investigate the contribution of nutrition to vitamin A and p-carotene supply in mother-infant pairs of multiparous births or births within short birth rates. Twenty-nine volimteers aged between 21 and 36 years were evaluated for 48 hours after delivery. In order to establish overall supply, retinol and p-carotene were determined in maternal plasma, cord blood, and colostrum via HPLC analysis. A food frequency protocol was obtained from all participants. Regardless of the high-to-moderate socioeconomic background, 27.6% of participants showed plasma retinol levels below 1.4 pmol/liter, which can be taken as borderline deficiency. In addition, 46.4% showed retinol intake <66% of RDA and 50.0% did not consume liver at all, although liver contributes as a main source for preformed retinol. Despite a high total carotenoid intake of 6.9 3.9mg/day, 20.7% of mothers showed plasma levels <0.5 pmol/liter p-carotene. 

---