Time (min)

Figure 4-6 Typical selected reaction monitoring profile from the analysis of capsaicinoids by LC—MS—MS (Reilly et al., 2001b). The identities of the peaks are octanoyl vanillamide (m/z 280-137), non-ivamide (m/z 294-137), capsaicin (m/z 306-137), and dihydrocapsaicin (m/z 308-137).

Capsaicinoids and toxicity

Many studies have been conducted on the most abundant capsaicinoids in the Capsicum pepper, capsaicin and dihydrocapsaicin. Capsaicin, the pungent phenolic compound of the Capsicum species, has shown a wide range of pharmacological properties, including antigenotoxic, antimu-tagenic and anticarcinogenic effects. Other studies, however, have shown it to be a tumour promoter and potential mutagen and carcinogen, resulting in capsaicin being termed as "a double edged sword" (Richeux et al., 1999; Singh et al., 2001).

Neurotoxicity, cytotoxicity and genotoxicity of capsaicinoids

The detection of painful stimuli occurs primarily at the peripheral terminals of specialized sensory neurons called nociceptors. The small diameter neurons transduce signals of a chemical, mechanical or thermal nature into action potentials and transmit their information to the central nervous system, ultimately eliciting a perception of pain and discomfort (Caterina and Julius, 2001). The localized desensitization of nociceptive afferents when capsaicin is applied in small doses has led to capsaicin being used as a therapeutic drug for pain relief (Minani et al., 2001). For example, capsaicin has been evaluated as an analgesic in a variety of neuropathic pain conditions, including postherapeutic neuralgia, painful diabetic neuropathy, osteroarthritis, the postsurgical pain syndrome and Guillain-Barre syndrome (Markovits and Gilhar, 1997; Szallasi and Blumberg, 1999). However, at increased concentrations, a capsaicin-associated burning sensation can negate the beneficial and therapeutic effects (Minani et al., 2001).

In order to determine the lethal toxic level of capsaicinoids and to extrapolate them to humans, many experiments were performed in mice, rats, guinea pig and rabbits. Pure capsaicin (16,000,000 SHU) was administered intravenously, subcutaneously in the stomach and applied topically until the death of the animals. The lethal toxic doses of capsaicin in mg per kg animal weight, were 0.56 for intravenous administration, 190 when consumed and 512 when applied topically. The probable cause of death in all cases was presumed to be respiratory paralysis. Guinea pig was the most sensitive to capsaicin, while rabbits were less sensitive. The acute toxicity of capsaicinoids as a food additive for humans was negligible. Moreover, if humans were as sensitive as mice, the acute fatal toxicity dose for a 70 kg person would be about 13 g of pure capsaicin which is very high regarding the Capsicum content and the quantity taken per one meal. Other studies have concluded that a person of 70 kg weight would have to consume nearly 2.5 L of Tabasco sauce to overdose and to become unconscious. People with few tastebuds in the mouth are not so bothered by the extreme heat. However, most of the people react very negatively to the super hot sauces, experiencing severe burning and sometimes blistering of the mouth and the tongue and immediate responses have included shortness of breath, fainting, nausea and spontaneous vomiting (Geppeti et al., 1988; Palecek et al., 1989; Midgren et al., 1992).

Many other studies on capsaicinoid toxicity have shown that capsaicin directly inhibits protein synthesis by competition with tyrosine in a cell-free system, as well as in a cell culture system according to the cell type and the extracellular concentration (Cochereau et al., 1996). Reports in the literature (Nagabhushan and Bhide, 1985; Lawso and Gannett, 1989; Morre et al., 1995) indicate that capsaicin and its metabolites are able to induce DNA damage and mutagenicity observed in bacteria and in vivo micronucleus formation in mice. In the presence of Cu (II) and molecular oxygen, capsaicin was reported to cause strand scission in DNA through an oxidative mechanism (Morre et al., 1995; Cochereau et al., 1996; Cochereau et al., 1997). For example, in Hela cells in culture, 100 jxM capsaicin was reported to induce DNA fragmentation and chromatin condensation (Morre et al., 1995).

Usage of capsaicinoids in self-defence weapons

An interesting application of capsaicinoids is their use in self-defence weapons. Defense sprays have become popular for both police use and personal protection. Most of them contain o-chlorobenzylidene, malononitrile, chloroacetophenone, oleoresin Capsicum or a combination of these ingredients. When applied topically, capsaicin produces a spontaneous inflammatory reaction in mucous membranes, and contact with eyes results in blepharospasm caused by irritation of the corneal nerves. Additional symptoms of eye contact include extreme burning heat, lacrimation, conjunctival edema and hyperemia (Gonzalez et al., 1993). In the nasal mucosa, capsaicin produces burning pain, sneezing and a dose-dependent serious discharge (Lundblad et al., 1984; Geppeti et al., 1988). Contact with skin produces a burning sensation, erythema without vesiculation, while its inhalation results in transitory bronchoconstriction, cough and retrosternal discomfort (Collier and Fuller, 1984; Fuller et al., 1985; Blanc et al., 1991; Midgren et al., 1992). Direct administration of extratracheal capsaicin aerosol in dogs resulted in apnea, brachycardia, hypotension, miosis and aqueous flare (Gonzalez et al., 1993).


Capsaicinoids are a natural group of alkaloids attracting an increasing interest for their use as food additives and for their pharmacological properties. Human intoxication risk with capsaicinoids after ingestion was evaluated to be very low, while its use in self-defence weapons with possible contact with eyes could result in acute blepharospam. Their isolation from the Capsicum species and their complete quantification still requires relatively sophisticated apparatus. However, permanent research for low cost determination continues.


AOAC Official Method 995.03- Capsaicinoids in capsicum and their extractives. Liquid chromatographic method. First action 1995. Bajaj, K. L. and Kaur, G. (1979) Colorimetric determination of capsaicin in capsicum fruits with the Folin-

Ciocalteu reagent. Mikrochim. Acta., 1, 81—86. Bernai, A. M., Calderon, A. A., Pedreno, M. A., Munoz, R., Barcelo, A. R. and Merino, F. C. (1993a) Capsaicin oxidation by peroxidase from Capsicum annuum (var. annuum) fruits. J. Agric. FoodChem., 41, 1041—1044. Bernai, A. M., Calderon, A. A., Pedreno, A. M., Barcelo, A. R. and Merino, C. F. (1993b) Dihydrocapsaicin oxidation by Capsicum annuum (var. annuum) peroxidase. J. Food Sa., 58(679), 611—613. Blanc, P., Luc, D. and Juarez, C. (1991) Cough in hot pepper workers. Chest, 99(1), 27-32. Caterina, M. J. and Julius, D. (2001) The vanilloid receptor: a molecular gateway to the pain pathway. Ann.

Rev. Neurosa., 24, 487-517. Cochereau, C., Sanchez, D., Bourhaoui, A. and Creppy, E. E. (1996) Capsaicin, a structural analog of tyrosine inhibits the aminoacylation of tRNAtyr. Toxicol. Appl. Pharmacol., 141, 133-137. Cochereau, C., Sanchez, D. and Creppy, E. E. (1997) Tyrosine prevents capsaicin-induced protein synthesis inhibition in cultured cells. Toxicology, 117, 133-139-Collier, J. and Fuller, R. (1984) Capsaicin inhalation in man and the effects of sodium cromoglycate. Br. J. Pharmacol., 81, 113-117.

Constant, H. L., Cordell, G. A., West, D. P. and Johnson J. H. (1995) Separation and quantification of capsaicinoids using complexation chromatography, j. Nat. Prod., 58(12), 1925-1928. Constant, H. L. and Cordell, G. A. (1996) Nonivamide, a constituent of capsicum oleoresin. J. Nat. Prod., 59(4), 425-426.

Cooper, T. H., Guzinski, J. A. and Fisher, C. (1991) Improved high-performance liquid chromatography method for the determination of major capsaicinoids in capsicum oleoresins._/. Agrtc. FoodChem., 39, 2253-2256.

Dicecco, J. J. (1976) Gas-liquid chromatographic determination of capsaicin. J. Assoc. Off. Anal. Chem., 59(1), 1-4.

Esteves, M. P., Girio, E. M., Amaral-Collaco, M. T., Andrade, M. E. and Empis, J. (1997) Characterization of starch from white and black pepper treated by ionizing radiation. Sciences des Aliments, 17, 289-298.

Fujiwake, H., Suzuki, T. and Iwai, K. (1982a) Intracellular distribution of enzymes and intermediates involved in the biosynthesis of capsaicin and its analogs in capsicum fruits. Agric. Biol. Chem., 46, 2685-2689.

Fujiwake, H., Suzuki, T. and Iwai. K. (1982b) Capsaicinoid formation in the protoplast from the placenta of capsicum fruits. Agric. Biol. Chem., 46, 2591-2595.

Fuller, R., Dixon, C. and Barnes, P. (1985) Bronchoconstrictor response to inhaled capsaicin in humans. J. Appl. Physiol, 58 (4), 1080-1084.

Fung, T., Jeffery, W. and Beveridge, A. D. (1982) The identification of capsaicinoids in tear-gas spray. J. Forensic Sci., 27, 812-821.

Geppeti, P., Fusco, B. and Marabine, S. (1988) Secretion, pain and sneezing induced the application of capsaicin to the nasal mucosa. Br. J. Pharmacol., 93, 509—514.

Gonzalez, G., Rubia, P. and Callar, J. (1993) Reduction of capsaicin-induced ocular pain and neurogenic inflammation by calcium antagonists. Invest. Ophthalmol. Vis. Sci., 34(12), 3329-3335.

Govindarajan, V. S. (1986) Ca/>jif»ff2-production, technology, and quality. Part III: chemistry of the color, aroma, and pungency stimuli. CRC Crit. Rev. Food Sci. Nutr., 24(3), 245-355.

Govindarajan, V. S., Narasimhan, S. and Dhanara, S. J. (1977) Evaluation of spices and oleoresin II. Pungency of Scoville Heat Units. A standardized procedure. J. FoodSci. Technol., 14, 28—34.

Hall, R. D., Holden, M. A. and Yeoman, M. M. (1987) The accumulation of phenylpropanoid and capsaicinoid compounds in cell cultures and whole fruit of the chilli pepper Capsicum frutescens. Mill. Plant Cell, Tiss. Org. Cult., 8, 163-176.

Hawer, W. S., Ha, J., Hwang, J. and Nam, Y. (1994) Effective separation and quantitative analysis of major heat principles in red pepper by capillary gas chromatography. Food Chem., 49, 99—103-

Howard, L. R., Talcott, S. T., Brenes, C. H. and Villalon, B. (2000) Changes in phytochemical and antioxidant activity of selected peppers cultivars (Capsicum species) as influenced by maturity. J. Agric. Food Chem., 48, 1713-1720.

Inoue, K., Sewalt, V. J. H., Balance, G. M., Ni, W., Sturzen, C. and Dixon, R. A. (1998) Developmental expression and substrate specifities of a,a-caffeic acid-3-0-methyltransferase and caffeoylcoenzyme A 3-0-methyltransferase in relation to lignification. Plant Physiol., 117, 761—770.

Iwai, K., Suzuki, T., Fujiwake, H. and Oha, S. (1979) Simultaneous microdetermination of capsaicin and its four analogues by using high-performance liquid chromatography and gas chromatography—mass spectrometry. J. Chromatogr., 172, 303—311.

Jurenitsch, J., David, M., Heresch, F. and Kubelka, W. (1979) Detection and identification of new pungent compounds in fruits of capsicum. Planta Med., 36, 61-67.

Jurenitsch, J. and Leinmuller, R. (1980) Quantification of nonylic acid vanillylamide and other capsaicinoids in the pungent principle of capsicum fruits and preparation by gas liquid chromatography on glass capillary columns (in German). J. Chromatogr., 189, 389-393.

Kosuge, S. and Furuta, M. (1970) Studies on the pungent principle of capsicum. Part XIV: Chemical constitution of the pungent principle. Agric. Biol. Chem., 34(2), 248-256.

Krajewska, A. M. and Powers, J. J. (1987) Gas chromatographic determination of capsaicinoids in green capsicum fruits. J. Assoc. Off. Anal. Chem. Int., 70(5), 926-928.

Laskaridou-Monnerville, A.(1999) Determination of capsaicinoid dihydrocapsaicin by micellar electroki-netic capillary chromatography and its application to various species of Capsicum, Solanaceae. J. Chromatogr. A, 838, 293-302.

Lawso, T. and Gannett, P. (1989) The mutagenicity of capsaicin and dihydrocapsaicin in V79 ceils. Cancer Lett., 49, 109-114.

Lee, K. R., Suzuki, T., Kobashi, M., Hasegawa, K. and Iwai, K. (1976) Quantitative micro analysis of capsaicin, dihydrocapsaicin, nordihydrocapsaicin using mass fragmentography. J. Chromatogr., 123, 119-128.

Lopez-Hernandez, J., Oruna-Concha, M.J., Simal-Lozano, J. M. E., Vazquez-Bianco, M. E. and GonzalezCastro, M. J. (1996) Chemical composition of padron peppers (Capsicum annuum L.) grown in Galicia (NW Spain). J. FoodChem., 57(4), 557-559.

Lundblad, L., Xiao Y. H. and Lundberg, J. (1984) Mechanisms for reflexive hypertension induced by local application of capsaicin and nicotine to the nasal mucosa. Acta. Physiol. Scand., 121, 277—282.

Machlin, L. J. (1995) Critical assessment of the epidemiologic data concerning the impact of antioxidant nutrients on cancer and cardiovascular disease. Crit. Rev. FoodSci. Nutr., 35, 41—50.

Manirakiza, P., Covaci, A. and Schepens, P. (1999) Solid-phase extraction and gas chromatography with mass spectrometric determination of capsaicin and some of its analogues from chilli peppers {Capsicum spp,)J. Assoc. Off. Anal. Chem. Int., 82(6), 1399-1405.

Manirakiza, P., Covaci, A. and Schepens, P. (2000) Single step clean up and GC-MS quantification of organochlorine pesticide residues in spice powder. Chromatographia, 52, 787—790.

Markovits, E. and Gilhar, A. (1997) Capsaicin: an effective topical treatment in pain. Int. J. Dermatol, 36, 401-404.

Markus, F., Daood, H. G., Kapitany, J. and Biacs, P. A. (1999) Change in the carotenoid and antioxidant content of spice red pepper (paprika) as a function of ripening and some technological factors. J. Agric. Food Chem., Al, 100-107.

Meilgaard, M., Civille, G. V. and Carr, B. T. (1987) Sensmy Evaluation Techniques, CRC Press, Boca Rotan, EL, Vol.2.

Midgren, B., Hansson, L. and Karlsson, J. (1992) Capsaicin-induced cough in humans. Am. Rev. Resp. Dis., 146(2), 347-351.

Minani, T., Bakoshi, S., Nakano, H., Mine, O., Muratami, T., Mori, H. and Ato, S. (2001) The effects of capsaicin cream on prostaglandin-induced allodynia. Anesth. Analg., 93, 419—423.

Morre, D. J., Chueh, P. J. and Morre, D. M. (1995) Capsaicin inhibits preferentially the NADH oxidase and growth of transformed cells in culture. Proc. Natl Acad. Sci. U.S.A., 92, 1831-1835.

Muller, H. (1997) Determination of the carotenoid content in selected vegetables and fruit by HPLC and photodiode array detection. Z. Lebensm. Unters. Forsch. A, 204, 88-94.

Murakami, K., Ito, M., Htay, H. H., Tsubouchi, R. and Yoshimo, M. (2001) Antioxidant effect of capsaicinoids on the metal-catalyzed lipid peroxidation. Biomed. Res. (Tokyo), 22(1), 15—17.

Nagabhushan, M. and Bhide, S. V. (1985) Mutagenicity of chilli extract and capsaicin in short-term tests. Environ. Mutagen., 7, 881-888.

Nakatani, N., Inatani, R., Ohta, H. and Nishioka, A. (1986) Chemical constituents of peppers (Piper spp.) and application to food preservation: naturally occurring antioxidative compounds. Environ. Health Perspect., 67, 135-142.

Neumann, D. (1966) On the biosynthesis of capsaicin. Naturwissenchaften., 53, 131-136.

Ochoa-Alejo, N. and Gomez-Peralta, J. E. (1998) Activity of enzymes involved in capsaicin biosynthesis in callus tissue and fruits of chilli peppers (Capsicum annuum I*.). J. Plant Physiol, 141, 147—152.

Palecek, F., Sant'ambrogio, G. and Sant'ambrogio, F. (1989) Reflex responses to capsaicin intravenous, aerosol, and intratracheal administration. J. Appl. Physiol., 67(4), 1428-1437.

Perucka, I. and Oieszek, W. (2000) Extraction and determination of capsaicinoids in fruit of hot pepper Capsicum annum. L. by spectrophotometry and high performance liquid chromatography. Food Chem., 71(2), 287-291.

Peusch, M., Muller-Seitz, E., Muller, M. P. A. and Anklam, E. (1997) Extraction of capsaicinoids from chillies (Capsicum frutescens L.) and paprika (Capsicum annuum L.) using supercritical fluids and organic solvents. Z. Lebensm. Unters. Forsch. A, 204, 351—355.

Ramos, P. J. J. (1979) Further study on the spectrophotometric determination of capsaicin. J. Assoc. Off. Anal. Chem., 62, 1168-1170.

Reilly, C. A., Crouch, D. J. and Yost, G. S. (2001a) Quantitative analysis of capsaicinoids in fresh peppers, oleoresin, capsicum, and pepper spray products. J. Forensic Sci., 46(3), 502-509.

Reilly, C. A., Crouch, D. J., Yost, G. S. and Fatah, A. A. (2001b) Determination of capsaicin, dihydrocapsaicin, and nonivamide in seif-defense weapons by liquid chromatography-mass spectrometry.

Richeux, F., Cascante, M., Ennamany, R., Saboureau, D. and Creppy, E. E. (1999) Cytotoxicity and geno-toxicity of capsaicin in human neuroblastoma cells SHSY-5Y. Arch. Toxicol, 73, 403-409-

Rico Avila, J. (1983) Cultivo del pimiento de Carne gruessa en invernadero. Ministerio de Agricultura, Pesca y Alimentación, Publicaciones de Extension Agraria, Madrid.

Rymal, K. S., Cosper, R. D. and Smith, D. A. (1979) Injection-extraction procedure for rapid determination of relative pungency in fresh Jalapeno peppers. J. Assoc. Off. Anal. Chem., 67, 658-659-

Saria, A., Lambeck, F. and Skofitsch, G. (1981) Determination of capsaicin in tissue and separation of capsaicin analogues by HPLC.J. Chromatogr., 208, 41—46.

Singh, S., Asad, S. F., Ahmad, A., Khan, N. U. and Hadi, S. M. (2001) Oxidative DNA damage by capsaicin and dihydrocapsaicin in the presence of Cu(II). Cancer Lett., 169, 139-146.

Sprague, J., Harrison, C., Rowbotham, D. J., Smart, D. and Lambert, D. G. (2001) Temperature-dependent activation of recombinant rat vanilloid VR1 receptors expressed in HEK293 cells by capsaicin and anandamide. Eur. J. Pharmacol., 423, 121—125.

Sukrasno, N. and Yeoman, M. M. (1993) Phenylpropanoid metabolism during growth and development of Capsicum frutescens. Phytochem., 32, 839-844.

Suzuki, T., Fujiwake, H. and Iwai, K. (1980) Intracellular localization of capsaicin and its analogues, capsaicinoids, in capsicum fruit. Microscopic investigation of the structure of the placenta of Capsicum annuum vox.annuum cv. Karayatsubusa. Plant Cell Physiol., 21, 33—37.

Szallasi, A. and Blumberg, P. M. (1999) Vanilloid (capsicum) receptor and mechanisms. Pharmacol. Rev. ,31, 139-212.

Thomas, B. V., Schreiber, A. A. and Weisskopf, C. P. (1998) Simple method for quantification of capsaicinoids in peppers using capillary gas chromatography. J. Agrie. Food Chem., 46, 2655-2663.

Todd, P. H., Besinger, M. G. and Biftu, T. (1977) Determination of pungency due to capsicum by gasliquid chromatography. J. Food Set., 42, 660—665.

Zewdie, Y. and Bosland, P. W. (2000) Capsaicinoid inheritance in an interspecific hybridization of Capsicum annuum X Capsicum chínense. J. Am. Soc. Hortic. Sci., 125(4), 448—453.

Zewdie, Y. and Bosland, P. W. (2001) Capsaicinoids profiles are not good chemotaxonomic indicators for capsaicin species. Biochem. Syst. EcoL, 29(2), 161—169-

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