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FATTY ACID COMPOSITION OF Capsicum GENUS PEPPERS

Composição de ácidos graxos em pimentas do gênero Capsicum

Abstracts

Fatty acids have a great metabolic and structural importance. Evaluation of fatty acid composition of peppers is still incomplete. Pulps and seeds from six varieties of the genus Capsicum were evaluated in this work with respect to their contents in fatty acids. A total of 25 different fatty acids, including some with odd number of carbons were identified in the samples. The most abundant fatty acids were palmitic (16:0), oleic (18:1n-9) and linoleic (18:2n-6) acids. The polyunsaturated:saturated fatty acid (PUFA/SFA) ratios for all peppers were high due to the elevated amounts of polyunsaturated acids, particularly linoleic acid. In the pulps, the omega-6/omega-3 ratios ranging from 1.28 to 4.33, were relatively adequate if one considers that ratios between 0.25 and 1.0 in the human diet are regarded as highly appropriate. In the seeds, the levels of omega-3 were very low whereas the levels of omega-6 were high, leading to very inadequate omega-6/omega-3 ratios ranging from 74.2 to 279.6. Principal component analysis (PCA) explained 93.49% of the total variance of the data. Considering the PUFA/SFA ratio and omega-6/omega-3 ratio, our data suggest that, among the peppers of the genus Capsicum evaluated in this work, the bell pepper and orange habanero pepper present the best nutritional characteristics concerning fatty acid composition.

Polyunsaturated:saturated ratio; omega-6/omega-3 ratio; multivariate analysis


Ácidos graxos tem grande importância metabólica e estrutural. Avaliação da composição de ácidos graxos em pimentas é ainda incompleta. Polpas e sementes de seis variedades de pimentas do gênero Capsicum foram avaliadas, neste trabalho, em relação a sua composição de ácidos graxos. Foram identificados 25 ácidos graxos diferentes nas amostras, incluindo alguns com número ímpar de átomos de carbono. Os ácidos graxos mais abundantes foram palmítico (16:0), oleico (18:1n-9) e linoleico (18:2n-6). As relações poli-insaturados:saturados foram elevadas para todas as amostras, em razão das elevadas quantidades de ácidos poli-insaturados, principalmente o ácido linoleico. Nas polpas as relações de omega-6/omega-3 variaram de 1,28 a 4,33, e foram avaliadas como relativamente adequadas, tendo em vista que, na dieta humana, relações entre 0,25 e 1,0 são consideradas altamente adequadas. Nas sementes, os níveis de n-3 foram muito mais baixos do que os níveis de n-6, que foram bem elevados, resultando em baixas proporções de omega-6/omega-3, variando entre 74,2-279,6. Análise de componentes principais (ACP) explicou 93,49% da variância total dos dados. Considerando as relações poli-insaturados: saturados e omega-6/omega-3, nossos dados sugerem que entre as pimentas do gênero Capsicum avaliadas, neste trabalho, o pimentão e a pimenta habanero laranja apresentaram as melhores características nutricionais em relação à composição de ácidos graxos.

Relação poli-insaturado:saturado; relação omega-6/omega-3; análises multivariadas


Introduction

Peppers (Capsicum spp.), which are grown worldwide, including Brazil, are used extensively as a natural food colorant and seasoning agent due to their attractive color, flavor, and taste (Reyes-Escogido et al., 2011REYES-ESCOGIDO, M.L.; GONZALEZ-MONDRAGON, E.G.; VAZQUEZ-TZOMPANTZI, E. Chemical and pharmacological aspects of capsaicin. Molecules. 16:1253-1270, 2011.). Capsicum spp are remarkable sources of antioxidant compounds, including capsaicinoids (Ochi et al., 2003OCHI, T. et al. Antioxidant activity of a new capsaicin derivative from Capsicum annuum. Journal of Natural Products. 66(8):1094-1096, 2003.; Topuz; Ozdemir, 2007TOPUZ, A.; OZDEMIR, F. Assessment of carotenoids, capsaicinoids and ascorbic acid composition of some selected pepper cultivars (Capsicum annum L.) grown in Turkey. Journal of Food Composition and Analysis. 20(7):596-602, 2007.) and phenolic compounds, particularly flavonoids (Materská; Perucka, 2005MATERSKÁ, M.; PERUCKA, I. Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L). Journal of Agricultural and Food Chemistry. 53(5):1750-1756, 2005.). The consumption of these components has potential health benefits due to their antioxidant activity, which may help to prevent inflammatory diseases and pathologies associated with oxidative damage, such as atherosclerosis and Alzheimer's disease (Kothari et al., 2010KOTHARI, S.L. et al. Chilli peppers-a review on tissue culture and transgenesis. Biotechnology Advances. 28(1):35-48, 2010.; Topuz; Ozdemir, 2007TOPUZ, A.; OZDEMIR, F. Assessment of carotenoids, capsaicinoids and ascorbic acid composition of some selected pepper cultivars (Capsicum annum L.) grown in Turkey. Journal of Food Composition and Analysis. 20(7):596-602, 2007.). Peppers of the genus Capsicum appear to possess anti-inflammatory, antioxidant, antiplatelet, antihypertensive, hypoglycemic, and hypocholesterolemic properties and demonstrated using in vitro and in vivo models. The main pungent component, capsaicin, has been used clinically for its analgesic and anti-inflammatory properties (Rezanka; Sigler, 2009REZANKA, T.; SIGLER, K. Odd-numbered very-long-chain fatty acids from the microbial, animal and plant kingdoms. Progress in Lipid Research. 48(3-4):206-238, 2009.; Srinivasan, 2013SRINIVASAN, K.; Biological Activities of Pepper Alkaloids. In RAMAWAT, K. G.; MÉRILLON, J. M. Natural Products. Berlin: Springer-Verlag, 2013, p. 1397.).

Lipids and fatty acids represent in general a small portion of the edible portion in peppers. However, they have a great metabolic and structural importance. The content of lipids is an important parameter of quality, considering that several bioactives and molecules responsible for the pericarp (pulp) colour are liposoluble. Additionally, the nutritional quality of lipids is dependent on its fatty acid composition (Martínez et al., 2006MARTÍNEZ, S. et al. Perfil de ácidos grasos de lagrasa de tres variedades de pimientos (Arnoia, Fresnode la Vega y los Valles-Benavente). Influencia del grado de maduración. Grasas y Aceites. 57(4):415-421, 2006.; Zaki et al., 2013ZAKI, N.; HASIB, A. et al. Assessment of color, capsaicinoids, carotenoids and fatty acids composition of paprika produced from Moroccan pepper cultivars (Capsicum annuum L.). Journal of Natural Sciences Research. 3(7):11-119, 2013.).

Evaluation of the fatty acid composition of peppers of the genus Capsicum is still incomplete and is limited to C. annuum (Jarret et al., 2013JARRET, R.L. et al. Seed oil content and fatty acid composition in a gene bank collection of Cucurbita moschata Duchesne and C. argyrosperma C. Huber. Plant Genetic Resources. 11(2):149-157, 2013.; Pérez-Gálvez et al., 1999PÉREZ-GÁLVEZ, A. et al. Fatty acid composition of two pepper varieties (Capsicum annuum L. cv. Jaranda y Jariza). Effect of drying process and nutritional aspects.Journal of the American Oil Chemists' Society. 76:205-208, 1999.). Thus, the objective of this work was to evaluate the fatty acid composition of six varieties of four species of peppers of the genus Capsicum.

Material and methods

The standards and reagents chloroform, methanol, standard methyl esters tricosanoic acid methyl ester (23:0) were purchased from Sigma-Aldrich Co (St Louis, USA).

Four species of the genus Capsicum, largely produced and consumed in Brazil, were used: C. annuum (two varieties, cayenne pepper and bell pepper), C. baccatum var. pendulum (known in Brazil as 'dedo de moça' pepper), C. chinense (two varieties, red habanero pepper and orange habanero pepper) and C. frutescens ('malagueta' pepper). They were all produced in the Northwest Paraná region, Southern Brazil (23º21' South latitude, 52º04' West longitude and 510 m altitude), between March and May 2013. Three different samples (at least 1 Kg) of each of these six varieties were purchased. All samples were at stage and conditions appropriate for human consumption (ripened fruits). Analyses were done immediately after purchase.

Pulps (pericarp) and seeds were manually separated and grounded in a blender. Lipids were extracted using a mixture of chloroform: methanol: water (45:30:25) (Bligh; Dyer, 1959BLIGH, E. G.; DYER, W.J. A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology. 37(8):911-917, 1959.).

To determine the fatty acid composition, the lipids were converted into fatty acid methyl esters (FAME) and were methylated (Hartman; Lago, 1973HARTMAN, L.; LAGO, R. C. A. Rapid preparation of fatty acid methyl from lipids. Laboratory practice. 22(6):474-476, 1973.). The FAME were separated using a gas chromatograph CP-3380 (Varian, USA) fitted with a flame ionization detector and a CP 7420-select Fame fused-silica capillary column (100 m x 0.25 mm x 0.25 μm cyanopropyl) (Souza et al., 2013SOUZA A. H. P. et al. Sacha inchi as potential source of essential fatty acids and tocopherols: multivariate study of nut and shell. Acta Scientiarum. Technology. 35(4):757-763, 2013.).

The retention times were compared to those of standard methyl esters (Sigma, USA). The fatty acids were quantified using tricosanoic acid methyl ester (Sigma, USA) as an internal standard (Joseph; Ackman, 1992JOSEPH, J.D.; ACKMAN, R. Capillary column gas chromatographic method for analysis of encapsulated fish oils and fish oil ethyl esters: collaborative study. Journal of American Oil Chemists' Society. 75(3):488-506, 1992.). The peak areas were determined with software Star 5.0 (Varian, USA) and the concentrations were expressed in mg g-1of total lipid.

The results are presented as the mean±standard deviation of three replicates of each experiment. A p-value of ≤0.05 was used to indicate significant differences between the mean values determined by analysis of variance (ANOVA) and Tukey´s test. The methodology of principal component analysis (PCA) was used to scrutinize the relationships between variables and standards. PCA was applied to separate the samples (pulps, seeds and varieties) according to the fatty acids. Firstly a matrix was build using the samples (n=12) and the responses (n=10), totalizing 120 points. The results obtained for each parameter were used as variable columns. Pulps and seeds of peppers were used as individual samples (lines). An autoscaled pre-treatment (z-score) of data was conducted to standardize the significant importance of all variables (Zielinski et al., 2014___________ ___. Chemical composition, sensory properties, provenance, and bioactivity of fruit juices as assessed by chemometrics: a critical review and guideline. Comprehensive reviews in food science and food safety. 13(3):300-316, 2014b.a). Analysis was based on linear correlations and variances were computed as sums of squares/(n-1). Finally, scatter plots was built for projecting all answers and standards (Zielinski et al., 2014___________ ___. Chemical composition, sensory properties, provenance, and bioactivity of fruit juices as assessed by chemometrics: a critical review and guideline. Comprehensive reviews in food science and food safety. 13(3):300-316, 2014b.b). All procedures were done by using STATISTICA 7.0 software (Stat-Soft Inc., Tulsa, OK, USA).

Results and discussion

Considering that the fatty acids are present in both pericarp (pulp) and seed, these parts were manually separated for evaluation of the distribution of those compounds. In all peppers the seeds were richer in total lipids than the pulps (Figure 1). The highest values were found in the seeds of 'dedo de moça' (10.17%) and cayenne (6.59%). The lowest values of total lipids were found in orange habanero (1.93%). Among pulps, the highest amounts of lipids were found in the 'malagueta' pepper. Peppers contain lipids that are qualitatively similar to those found in plants in general, but their fruits are considered as poor in lipids, normally less than 2% (Asilbekova, 2003ASILBEKOVA, D.T. Lipids from Capsicum annum seeds. Chemistry of Natural Compounds. 39(6):528-530, 2003.; Bosland; Votava, 2012BOSLAND, P.; VOTAVA, E. Peppers - vegetable and spice capsicums. Wallingford, England: CABI, 2012, 2th ed, 230p.).

Figure 1:
Total lipids of pulps and seeds of peppers of the genus Capsicum.

The fatty acid composition of the pulps and seeds of peppers is shown in Tables 1 and 2. A total of 25 different fatty acids, including some with odd number of carbons, were identified. Such odd fatty acids occur in small quantities in vegetables and are receiving attention only now because of the technical difficulties inherent in their detection and identification ( Diedrich; Henschel, 1990DIEDRICH.M.; HENSCHEL, K.P. The natural occurrence of unusual fatty acids - Part I. Odd numbered fatty acids. Molecular Nutrition& Food Research. 34(10):935-943, 1990.; Martínez et al., 2006MARTÍNEZ, S. et al. Perfil de ácidos grasos de lagrasa de tres variedades de pimientos (Arnoia, Fresnode la Vega y los Valles-Benavente). Influencia del grado de maduración. Grasas y Aceites. 57(4):415-421, 2006.; Rezanka; Sigler, 2009REZANKA, T.; SIGLER, K. Odd-numbered very-long-chain fatty acids from the microbial, animal and plant kingdoms. Progress in Lipid Research. 48(3-4):206-238, 2009.).

Table 1:
Fatty acid composition of pulps of peppers.
Table 2:
Fatty acid composition of seeds of peppers.

As in the majority of plants, the most abundant fatty acids in the peppers were palmitic (16:0), oleic (18:1n-9) and linoleic (18:2n-6) acids (Table 1 and 2). The last one was the most abundant in all seeds and pulps. Its contents range from 325.35 to 209.03 mg per 100 g of pulp (Table 1) and 697.29 to 669.66 mg per 100 g of seed (Table 2). In general, the major fatty acids in pulps and seeds are the unsaturated ones (apart from palmitic acid, which is the precursor of the others). As the 18:2 and 18:3 fatty acids are synthesized preferentially, they probably constitute the appropriate membrane lipids in this stage of fruit ripening (Pérez-Gálvez et al., 1999PÉREZ-GÁLVEZ, A. et al. Fatty acid composition of two pepper varieties (Capsicum annuum L. cv. Jaranda y Jariza). Effect of drying process and nutritional aspects.Journal of the American Oil Chemists' Society. 76:205-208, 1999.). It is interesting to note that linolenic acid (18:3n-3) is found in high amounts in pulps, but it is almost absent in seeds. This phenomenon has already been reported for other species of the genus Capsicum (Pérez-Gálvez et al., 1999PÉREZ-GÁLVEZ, A. et al. Fatty acid composition of two pepper varieties (Capsicum annuum L. cv. Jaranda y Jariza). Effect of drying process and nutritional aspects.Journal of the American Oil Chemists' Society. 76:205-208, 1999.). The reasons and implications for this particularity cannot be inferred from the hitherto available data.

The PUFA:SFA ratios for all peppers were high due to the elevated amounts of polyunsaturated acids, particularly linoleic acid (Table 3). Foods with high polyunsaturated/saturated fatty acid ratio (PUFA/SFA) are considered beneficial for health, especially because they contribute for the reduction of fat body and total cholesterol (Simopoulos, 2011_________________. Evolutionary aspects of the diet: the Omega-6/omega-3 ratio and the brain. Molecular Neurobiology. 44(2):203-215, 2011.). Furthermore, PUFA, especially omega-3, when incorporated into the cells can act by modulating several metabolic and signalling pathways and by exerting protective effects against inflammatory and tumoral events (Alexander, 1998ALEXANDER, J.W. Immunonutrition: The role of ω-3 fatty acids. Nutrition. 14(7-8):627-633, 1998.; Arrigo et al., 2004ARRIGO, M. D. et al. Production of n-3 fatty acid enriched pork liver pâté. LWT - Food Science and Technology. 37(6):585-591, 2004.; Gladine et al., 2012GLADINE, C. et al. Increasing intake of long chain omega 3 PUFA enhances lipoperoxidation and modulates hepatic gene expression in a dose-dependent manner. British Journal of Nutrition 107(9):1254-1273, 2012.; Rose; Connolly, 1999ROSE, D.P.; CONNOLLY, J.M. Omega-3 fatty acids as cancer chemopreventive agents. Pharmacology and Therapeutics. 83(3):217-244, 1999.).

Table 3:
Sums and ratios of fatty acid.

The omega-6:omega-3 ratios in the pulps, from 1.28 to 4.33 (Table 3), are relatively adequate if one considers that ratios between 0.25 and 1.0 in the human diet are regarded as highly appropriate. In the seeds, the levels of n-3 were very low whereas the levels of n-6 were high, leading to very inadequate ratios ranging from 74.2 to 279.6. Considering that foods with a good omega-6:omega-3 ratio can act by preventing inflammatory processes and diseases, the ingestion of pulps of peppers can generate more health benefits than the ingestion of seeds (Malacrida, 2010MALACRIDA, C.R. Characterization of oils extracted from fruit seeds: fatty acid composition, tocopherols and carotenoids. Revista Instituto Adolfo Lutz. 69(1):144, 2010.; Simopoulos, 2002SIMOPOULOS, A.P. The importance of the ratio of omega-6/omega-3 essential fatty acids. Biomedicine and Pharmacotherapy. 56(8):365-379, 2002.). To our knowledge, this is the first study to evaluate the omega-6/omega-3 ratio in peppers.

The application of univariate methods to a large number of variables and/or samples may not be appropriate for understanding the data structure and interrelationships. Therefore, the use of PCA can be an alternative because it allows to construct 2-D or 3-D graphs which facilitate understanding of similarities or differences (Zielinski et al., 2014___________ ___. Chemical composition, sensory properties, provenance, and bioactivity of fruit juices as assessed by chemometrics: a critical review and guideline. Comprehensive reviews in food science and food safety. 13(3):300-316, 2014b.a). Figure 2 illustrates the relationships of the samples with their principal fatty acid contents. PCA explained 93.49% of the total variance of the data, where PC1 explaining up 81.74% and PC2 11.75%. The samples were separated between seeds (left side) and pulps (right side). The seeds were related to the highest levels of linoleic acid, n-6, PUFA, as well as the PUFA:SFA ratio and the n-6:n-3 ratio, whereas the pulps showed higher contents of palmitic acid, oleic acid, MUFA, SFA and n-3. Among all pepper pulps, orange habanero and bell pepper pulps showed the higher levels of n-3.

Figure 2:
Scatter plots PC1 vs. PC2 on the main sources of variability among peppers samples. (A) Scores and (B) Loadings plots.

Conclusions

All peppers presented high values of PUFA, generally believed to be beneficial for health. Considering the PUFA:SFA and the n-6:n-3 ratios, our data suggest that, among the peppers of the genus Capsicum evaluated in this work, the bell pepper and the orange habanero pepper present the best nutritional characteristics with respect to their fatty acid composition.

Acknowledgements

This study was supported by Universidade Estadual de Maringá (UEM), Universidade Tecnológica Federal do Paraná (UTFPR), Conselho Nacional de Pesquisa (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nivel Superior (CAPES).

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  • BLIGH, E. G.; DYER, W.J. A rapid method of total lipid extraction and purification. Canadian Journal of Biochemistry and Physiology. 37(8):911-917, 1959.
  • BOSLAND, P.; VOTAVA, E. Peppers - vegetable and spice capsicums. Wallingford, England: CABI, 2012, 2th ed, 230p.
  • DIEDRICH.M.; HENSCHEL, K.P. The natural occurrence of unusual fatty acids - Part I. Odd numbered fatty acids. Molecular Nutrition& Food Research. 34(10):935-943, 1990.
  • GLADINE, C. et al. Increasing intake of long chain omega 3 PUFA enhances lipoperoxidation and modulates hepatic gene expression in a dose-dependent manner. British Journal of Nutrition 107(9):1254-1273, 2012.
  • HARTMAN, L.; LAGO, R. C. A. Rapid preparation of fatty acid methyl from lipids. Laboratory practice. 22(6):474-476, 1973.
  • JARRET, R.L. et al. Seed oil content and fatty acid composition in a gene bank collection of Cucurbita moschata Duchesne and C. argyrosperma C. Huber. Plant Genetic Resources. 11(2):149-157, 2013.
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  • MARTÍNEZ, S. et al. Perfil de ácidos grasos de lagrasa de tres variedades de pimientos (Arnoia, Fresnode la Vega y los Valles-Benavente). Influencia del grado de maduración. Grasas y Aceites. 57(4):415-421, 2006.
  • MATERSKÁ, M.; PERUCKA, I. Antioxidant activity of the main phenolic compounds isolated from hot pepper fruit (Capsicum annuum L). Journal of Agricultural and Food Chemistry. 53(5):1750-1756, 2005.
  • OCHI, T. et al. Antioxidant activity of a new capsaicin derivative from Capsicum annuum. Journal of Natural Products. 66(8):1094-1096, 2003.
  • PÉREZ-GÁLVEZ, A. et al. Fatty acid composition of two pepper varieties (Capsicum annuum L. cv. Jaranda y Jariza). Effect of drying process and nutritional aspects.Journal of the American Oil Chemists' Society. 76:205-208, 1999.
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  • REZANKA, T.; SIGLER, K. Odd-numbered very-long-chain fatty acids from the microbial, animal and plant kingdoms. Progress in Lipid Research. 48(3-4):206-238, 2009.
  • ROSE, D.P.; CONNOLLY, J.M. Omega-3 fatty acids as cancer chemopreventive agents. Pharmacology and Therapeutics. 83(3):217-244, 1999.
  • SIMOPOULOS, A.P. The importance of the ratio of omega-6/omega-3 essential fatty acids. Biomedicine and Pharmacotherapy. 56(8):365-379, 2002.
  • _________________. Evolutionary aspects of the diet: the Omega-6/omega-3 ratio and the brain. Molecular Neurobiology. 44(2):203-215, 2011.
  • SOUZA A. H. P. et al. Sacha inchi as potential source of essential fatty acids and tocopherols: multivariate study of nut and shell. Acta Scientiarum. Technology. 35(4):757-763, 2013.
  • SRINIVASAN, K.; Biological Activities of Pepper Alkaloids. In RAMAWAT, K. G.; MÉRILLON, J. M. Natural Products. Berlin: Springer-Verlag, 2013, p. 1397.
  • TOPUZ, A.; OZDEMIR, F. Assessment of carotenoids, capsaicinoids and ascorbic acid composition of some selected pepper cultivars (Capsicum annum L.) grown in Turkey. Journal of Food Composition and Analysis. 20(7):596-602, 2007.
  • ZAKI, N.; HASIB, A. et al. Assessment of color, capsaicinoids, carotenoids and fatty acids composition of paprika produced from Moroccan pepper cultivars (Capsicum annuum L.). Journal of Natural Sciences Research. 3(7):11-119, 2013.
  • ZIELINSKI, A.A.F. et al. A comparative study of the phenolic compounds and the in vitro antioxidant activity of different Brazilian teas using multivariate statistical techniques. Food Research International. 60:246-254, 2014a.
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Publication Dates

  • Publication in this collection
    Jul-Ago 2015

History

  • Received
    19 Feb 2015
  • Accepted
    27 Apr 2015
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