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Capsicum--production, technology, chemistry, and quality. Part III. Chemistry of the color, aroma, and pungency stimuli.

The spice capsicum, the fruits of the genus Capsicum (Family Solanaceae), is a very popular food additive in many parts of the world, valued for the important sensory attributes of color, pungency, and aroma. A large number of varieties are widely cultivated and traded. The characteristic carotenoids of the bright red paprika and cayenne-type chillies, the high character impact aroma stimuli, the methoxy pyrazine of green bell capsicum, the esters of ripe tabasco and the highly potent pungency stimuli, and the capsaicinoids of African and other Asian varieties of chillies, have been of great interest to chemists and biochemists. Research workers in other disciplines such as genetics and breeding, agriculture, and technology have been interested in this spice to develop new varieties with combinations of different optimal levels of the stimuli for the sensory attributes and to maximize production of storable products for specific end uses. Physiologists have been intensely studying the action of the highly potent pungency stimuli and social psychologists the curious aspect of growing acceptance and preference for the initially unacceptable pungency sensation. In the sequential review of all these aspects of the fruit spice Capsicum, the earlier two parts covered history, botany, cultivation and primary processing, and processed products, standards, world production, and trade. In Part III, the chemistry, the compositional variations, synthesis and biosynthesis of the functional components, the carotenoids, the volatiles, and the capsaicinoids are comprehensively reviewed.

Capsaicin↗

Capsicum--production, technology, chemistry, and quality. Part V. Impact on physiology, pharmacology, nutrition, and metabolism; structure, pungency, pain, and desensitization sequences.

The spice Capsicum is the fruit of the cultivated species of the genus Capsicum (family, Solanaceae), C. annuum principally, and C. frutescens L. to a lesser extent. A third variety of C. annuum var. annuum fruits, the large-sized, fleshy bell capsicum is used as a fresh vegetable and valued for its aroma, color, and crisp texture, but with no pungency. This variety is not considered in this series of reviews covering primary processing, production, international trade, chemistry, and biochemistry of functional components--the red keto carotenoids, the aromatic volatiles and the pungent capsaicinoids in Parts I to III. The valid qualitative aspects correlating the specific components of capsicum and their sensory responses are critically covered in Part IV. In this the concluding part of the series of reviews, the significant preference of the spice for initially evoking an aversive response, its potent physiological and pharmacological effects, and the aspects of structure-activity relationships of the pungent stimuli of the capsaicinoids are reviewed. The beneficial effects particularly associated with long usage by some ethnic groups and its safe consumption levels, with a critical review of the studies on the gastrointestinal tract, the cardiovascular system, the sensory system, thermoregulation, nutritional impacts, and an overview of the five series is also detailed.

Animals↗

Unilateral incompatibility in Capsicum (Solanaceae): occurrence and taxonomic distribution.

BACKGROUND AND AIMS: Unilateral incompatibility (UI) occurs when pollinations between species are successful in one direction but not in the other. Self-incompatible (SI) species frequently show UI with genetically related, self-compatible (SC) species, as pollen of SI species is compatible on the SC pistil, but not vice versa. Many examples of unilateral incompatibility, and all those which have been studied most intensively, are found in the Solanaceae, particularly Lycopersicon, Solanum, Nicotiana and Petunia. The genus Capsicum is evolutionarily somewhat distant from Lycopersicon and Solanum and even further removed from Nicotiana and Petunia. Unilateral incompatibility has also been reported in Capsicum; however, this is the first comprehensive study of crosses between all readily available species in the genus. METHODS: All readily available (wild and domesticated) species in the genus are used as plant material, including the three genera from the Capsicum pubescens complex plus eight other species. Pollinations were made on pot-grown plants in a glasshouse. The number of pistils pollinated per cross varied (from five to 40 pistils per plant), depending on the numbers of flowers available. Pistils were collected 24 h after pollination and fixed for 3-24 h. After staining, pistils were mounted in a drop of stain, squashed gently under a cover slip and examined microscopically under ultra-violet light for pollen tube growth. KEY RESULTS: Unilateral incompatibility is confirmed in the C. pubescens complex. Its direction conforms to that predominant in the Solanaceae and other families, i.e. pistils of self-incompatible species, or self-compatible taxa closely related to self-incompatible species, inhibit pollen tubes of self-compatible species. CONCLUSIONS: Unilateral incompatibility in Capsicum does not seem to have arisen to prevent introgression of self-compatibility into self-incompatible taxa, but as a by-product of divergence of the C. pubescens complex from the remainder of the genus.

Capsicum↗

Molecular mapping of the C locus for presence of pungency in Capsicum.

Pungency owing to the presence of capsaicinoids is a unique character of pepper (Capsicum spp.). Capsaicinoids are produced in the placenta and it has long been known that a single dominant gene, C, is required for pungent genotypes to produce capsaicinoids. We mapped C to pepper chromosome 2 in a cross between a pungent Capsicum frutescens wild accession and a non-pungent Capsicum annuum bell pepper. This position confirmed results from earlier studies. The RFLP marker TG 205 cosegregated with C and two additional RFLP markers were also located within 1 cM. The recessive allele at the C locus is used in breeding programs around the world focused on very diverse germplasm, hence any of these tightly linked markers may be of value as potential sources of useful markers for marker-assisted selection. To demonstrate this point, we developed a PCR-based CAPS (cleaved amplified polymorphic sequence) marker linked to C using the sequence of the Capsicum fibrillin gene located 0.4 cM from C. The use of molecular markers for high-throughput screening for the c allele in pepper breeding programs is discussed.

Capsaicin↗

fs3.1: a major fruit shape QTL conserved in Capsicum.

fs3.1 is a major fruit shape (defined as the ratio of fruit length to fruit width) quantitative trait locus (QTL) originally detected in an intraspecific cross of Capsicum annuum between the blocky and elongated-fruited inbreds 'Maor' and 'Perennial', respectively. In addition to increasing fruit shape index, the 'Perennial' allele at fs3.1 increased fruit elongation and decreased fruit width and pericarp thickness. We verified the effect of fs3.1 in backcross inbred lines (BILs) derived from crossing 'Perennial' with 'Maor' and with a second blocky-type inbred line of C. annuum. To determine the effect of the fs3.1 region in additional Capsicum species, we constructed an advanced backcross population from the cross of 'Maor' and the oval-fruited Capsicum frutescens BG 2816 and an F2 of the introgression line IL 152 that contains an introgression of the fs3.1 region from Capsicum chinense PI 152225. QTLs for fruit shape, fruit width, and pericarp thickness, but not for fruit length, were detected in both crosses, indicating the conservation of the fs3.1 region as a QTL affecting fruit shape in pepper. We also tested tomato (Lycopersicon spp.) introgression lines containing the corresponding fs3.1 region from L. pennellii and L. hirsutum, but we did not detect a significant fruit shape QTL in these lines. The effect of fs3.1 on the growth of fruit dimensions varied with the genetic background. By measuring the length and width of ovaries and fruits of near-isogenic C. annuum lines that differ in fs3.1 during fruit development, we determined that fs3.1 controls shape predominantly by increasing the growth rate of the longitudinal axis in the first 2 weeks after pollination. However, in the crosses of C. annuum with C. frutescens and C. chinense, fs3.1 predominantly exerted its effect on the width dimension.

Capsicum↗

Synergistic Capsicum-tea mixtures with anticancer activity.

We have demonstrated a synergy between a decaffeinated green tea concentrate and a vanilloid-containing Capsicum preparation obtained commercially. At a ratio of 25 parts green tea concentrate to 1 part Capsicum preparation, the resultant product exhibited efficacy in the killing of cancer cells in culture 100-times that of green tea on a weight basis. These studies were guided by assays of the putative catechin-vanilloid target protein tNOX, a cell surface growth-related enzymatic activity specific to cancer. The activity of the protein target was inhibited by the tea catechins and the Capsicum vanilloids. As with growth, the tea and Capsicum preparations evaluated were synergistic in their inhibition of the target enzymatic activity.

Antineoplastic Agents, Phytogenic↗

Quantitative analysis of capsaicinoids in fresh peppers, oleoresin capsicum and pepper spray products.

Liquid chromatography-mass spectrometry was used to identify and quantify the predominant capsaicinoid analogues in extracts of fresh peppers, in oleoresin capsicum, and pepper sprays. The concentration of capsaicinoids in fresh peppers was variable. Variability was dependent upon the relative pungency of the pepper type and geographical origin of the pepper. Nonivamide was conclusively identified in the extracts of fresh peppers, despite numerous reports that nonivamide was not a natural product. In the oleoresin capsicum samples, the pungency was proportional to the total concentration of capsaicinoids and was related by a factor of approximately 15,000 Scoville Heat Units (SHU)/microg of total capsaicinoids. The principle analogues detected in oleoresin capsicum were capsaicin and dihydrocapsaicin and appeared to be the analogues primarily responsible for the pungency of the sample. The analysis of selected samples of commercially available pepper spray products also demonstrated variability in the capsaicinoid concentrations. Variability was observed among products obtained from different manufacturers as well as from different product lots from the same manufacturer. These data indicate that commercial pepper products are not standardized for capsaicinoid content even though they are classified by SHU. Variability in the capsaicinoid concentrations in oleoresin capsicum-based self-defense weapons could alter potency and ultimately jeopardize the safety and health of users and assailants.

Capsaicin↗

The role of some weeds as hosts of Capsicum viruses in the rift valley parts of Ethiopia.

Hot pepper (Capsicum annuum. L.) is a traditionally and economically important spice crop in Ethiopia. Chronic infection with viruses is reported as major constraint, that often force farmers to ban its production. However, epidemiological studies on these viruses are lacking in the country. Knowledge of the weed flora as virus reservoir is essential to reduce the spread of plant viruses. The potential role of some frequently and abundantly occurring weed species as host plants for Capsicum viruses was therefore investigated. Surveys were conducted during the 2000/2001 cropping- and off-season. Samples of various weed species revealing virus symptoms were collected from different districts. Collection was done from inside and at the border of pepper cropped and fallow fields. It was also performed along road sides and irrigation channels. The identity of the collected isolates was established based on serology, electron microscopy and host range studies. Moreover, seeds were collected from natural populations of the most prevalent weeds, namely Datura stramonium and Nicandra physalodes. They were grown under glass house conditions and tested for their ability to host viruses. To verify their role further, healthy seedlings from these weeds were artificially inoculated with all viruses isolated from weeds and Capsicum leaf samples. Natural infection of different Potyviruses with an average incidence of 33% in the weed samples were the most dominant. In addition, the occurrence of Cucumber mosaic virus, Tomato mosaic virus and Potato virus X in these weeds was demonstrated. However, none of the isolated viruses was found to be seed borne in the tested weed species and pepper crop (var.Mrkofanna) under glass house conditions. Recently, there was an explosion in population size of some solanaceous weeds, particularly Datura and Nicandra species in the area. The presence of infected weeds throughout the year means, that they are an important reservoir and source for secondary spread. It is suggested therefore, that a rigorous weed control is an important step to reduce the incidence of viruses infecting Capsicum in the area.

Capsicum↗

[Analysis of microsatellite loci of the chloroplast genome in the genus Capsicum (Pepper)].

Six plastome microsatellites were examined in 43 accessions of the genus Capsicum. In total, 33 allelic variants were detected. A specific haplotype of chloroplast DNA was identified for each Capsicum species. Species-specific allelic variants were found for most wild Capsicum species. The highest intraspecific variation was observed for the C. baccatum plastome. Low cpDNA polymorphism was characteristic of C. annuum: the cpSSRs were either monomorphic or dimorphic. The vast majority of C. annuum accessions each had alleles of one type. Another allele type was rare and occurred only in wild accessions. The results testified again to genetic conservation of C. annuum and especially its cultivated forms. The phylogenetic relationships established for the Capsicum species on the basis of plastome analysis were similar to those inferred from the morphological traits, isozyme patterns, and molecular analysis of the nuclear genome.

Base Sequence↗

Effects of capsicum fruit on theophylline absorption and bioavailability in rabbits.

Absorption and bioavailability of theophylline from a sustained-release gelatin capsule were investigated in 10 male rabbits after oral administration (20 mg/kg), with and without a ground capsicum fruit suspension. Comparison of pharmacokinetic parameters showed that the concomitant absorption of capsicum increases areas under plasma curves (from 86.06 +/- 9.78 mg H/liter to 138.32 +/- 17.27 mg H/liter, P less than 0.001), peak plasma levels (from 6.65 +/- 0.76 to 8.78 +/- 0.98 mg/liter, P less than 0.01), and mean residence times (from 14.94 +/- 2.97 to 20.98 +/- 5.75 H, P less than 0.001). A second administration of the capsicum suspension, 11 hours after dosing, produced a new rise of theophylline plasma levels in every rabbit. The variations in pharmacokinetic and bioavailability parameters are discussed in accordance with the mechanisms of action of capsaicin, an active compound present in capsicum fruit.

Absorption↗

Prevention of PONV by acustimulation with capsicum plaster is comparable to ondansetron after middle ear surgery.

PURPOSE: To compare the efficacy of stimulation of P6 acupoint with capsicum plaster in comparison with iv ondansetron for the prevention of postoperative nausea and vomiting (PONV). METHODS: 120 patients of either sex, ASA I-II, undergoing elective middle ear surgeries under general anesthesia were included in this randomized, prospective, double-blinded and placebo-controlled study. The anesthetic technique was standardized. Patients were divided into three groups. Group I was the control group. Capsicum plaster (1 x 1 cm) was affixed at the P6 acupoint on both forearms 30 min before induction of anesthesia in patients of Group II. Patients of Groups I and III received an inactive adhesive plaster at the same site. Ondansetron 4 mg iv was given to patients of Group III at the end of surgery and the rest of the patients received a placebo. The plasters were removed six hours after transferring the patients to the postoperative unit. Criteria were fixed for the administration of rescue antiemetics (ondansetron 4 mg iv). PONV and the requirement for rescue antiemetics were recorded by a blinded observer. RESULTS: The incidence of PONV and the requirement for rescue antiemetics were significantly lower in both the acustimulation and ondansetron groups at six hours. At 24 hr there was a reduction in the requirement for rescue medication in the ondansetron group. CONCLUSION: Stimulation of the P6 acupoint with capsicum plaster is an effective method for prevention of PONV after middle ear surgery and its efficacy is comparable to ondansetron for the first six hours after surgery.

Acupuncture Therapy↗

Allele-specific CAPS markers based on point mutations in resistance alleles at the pvr1 locus encoding eIF4E in Capsicum.

Marker-assisted selection has been widely implemented in crop breeding and can be especially useful in cases where the traits of interest show recessive or polygenic inheritance and/or are difficult or impossible to select directly. Most indirect selection is based on DNA polymorphism linked to the target trait, resulting in error when the polymorphism recombines away from the mutation responsible for the trait and/or when the linkage between the mutation and the polymorphism is not conserved in all relevant genetic backgrounds. In this paper, we report the generation and use of molecular markers that define loci for selection using cleaved amplified polymorphic sequences (CAPS). These CAPS markers are based on nucleotide polymorphisms in the resistance gene that are perfectly correlated with disease resistance, the trait of interest. As a consequence, the possibility that the marker will not be linked to the trait in all backgrounds or that the marker will recombine away from the trait is eliminated. We have generated CAPS markers for three recessive viral resistance alleles used widely in pepper breeding, pvr1, pvr1 (1), and pvr1 (2). These markers are based on single nucleotide polymorphisms (SNPs) within the coding region of the pvr1 locus encoding an eIF4E homolog on chromosome 3. These three markers define a system of indirect selection for potyvirus resistance in Capsicum based on genomic sequence. We demonstrate the utility of this marker system using commercially significant germplasm representing two Capsicum species. Application of these markers to Capsicum improvement is discussed.

Alleles↗

Specific synthesis of 5,5'-dicapsaicin by cell suspension cultures of capsicum annuum var. annuum (chili Jalapeño chigol) and their soluble and NaCl-extracted cell wall protein fractions.

HPLC-UV, (1)H NMR, (13)C NMR, and (1)H-(1)H COSY analyses revealed that exogenous capsaicin was specifically converted into 5,5'-dicapsaicin by both cell suspension cultures of Capsicum annuum var. annuum (chili Jalapeño chigol) and their soluble and NaCl-extracted cell wall protein fractions under oxidative conditions. In cell suspension cultures 5,5'-dicapsaicin was found only in biomass of capsaicin-fed cultures. This compound has not been detected before either in fresh fruits or in in vitro cultures of Capsicum. The transformation of capsaicin by different protein fractions revealed that most of the enzymatic activity was located in the NaCl-extracted, or ionic cell wall bound, protein, and that it was strictly dependent on H(2)O(2). These results might in part explain some previously described features of capsaicin production by in vitro cultures of Capsicum. The implications of the results regarding the catabolism of capsaicinoids are discussed.

Capsaicin↗

Influence of 8-methyl-nonenoic acid on capsaicin biosynthesis in in-vivo and in-vitro cell cultures of Capsicum spp.

Capsaicin is a bioactive molecule synthesized by enzymatic (putative capsaicin synthase) condensation of vanillylamine, a phenyl propanoid intermediate with 8-methyl-nonenoic acid, a fatty acid derivative from leucine/valine pathway. Analysis of levels of 8-methyl-nonenoic acid and phenyl propanoid intermediates in high, medium, and low pungent Capsicum genotypes revealed that the 8-methyl-nonenoic acid pool plays a crucial role in determining the efficacy of capsaicin levels. Cerulenin-mediated inhibition of 8-methyl-nonenoic acid synthesis decreased the capsaicin biosynthesis in Capsicum cell suspension cultures. Similarly amino oxy acetate inhibited vanillylamine synthesis but failed to reduce capsaicin production. The mRNA transcript analysis of keto acyl synthase (KAS), a crucial enzyme involved in 8-methyl-nonenoic acid and an amino transferase (AMT), involved in vanillylamine biosynthesis was studied. The mRNA transcript analysis revealed the progressive developmental expression of KAS gene in the placenta during the ontogeny of the fruit, whereas AMT transcripts levels did not show significant differences. Hence, the study demonstrates the influence of 8-methyl-nonenoic acid and its possible regulatory role in capsaicin biosynthesis in Capsicum spp.

Capsaicin↗

Purification and structure determination of glucosides of capsaicin and dihydrocapsaicin from various Capsicum fruits.

Two new glucosides, capsaicin-beta-D-glucopyranoside (1) and dihydrocapsaicin-beta-D-glucopyranoside (2), were discovered in the fruit of the Capsicum annuum cultivar 'High Heat'. They were sequentially purified by acetone extraction, n-hexane extraction, and acetonitrile extraction, followed by medium-pressure liquid chromatography and high-performance liquid chromatography performed on an octadecylsilane column. Their chemical structures were elucidated by proton nuclear magnetic resonance, carbon nuclear magnetic resonance, and hydrolysis with alpha- and beta-glucosidases. The glucosides were also detected in various pungent cultivars of C. annuum, Capsicum frutescens, and Capsicum chinense by liquid chromatography-mass spectrometry. However, they were not detected in nonpungent cultivars of C. annuum. Furthermore, a positive correlation was observed between the quantity of the capsaicinoids, capsaicin, and dihydrocapsaicin and their glucosides.

Capsaicin↗

Determination of capsaicin and dihydrocapsaicin in capsicum fruits by liquid chromatography-electrospray/time-of-flight mass spectrometry.

A simple, highly selective, sensitive, and reproducible liquid chromatography-electrospray ionization/time-of-flight mass spectrometry method has been developed for the direct and simultaneous determination of capsaicin and dihydrocapsaicin in Capsicum fruit extracts. Capsaicin and dihydrocapsaicin are the two major members of the so-called capsaicinoid family, which includes other minor analogues, and usually account for at least 90% of the pungency trait in Capsicum fruits. Chromatographic separation of capsaicin and dihydrocapsaicin was achieved with a reversed-phase chromatography column, using a gradient of methanol and water. Quantification was done using as an internal standard (4,5-dimethoxybenzyl)-4-methyloctamide, a synthetic capsaicin analogue not found in nature. Analytes were base-peak resolved in less than 16 min, and limits of detection were 20 pmol for capsaicin and 4 pmol for dihydrocapsaicin. The intraday repeatability values were lower than 0.5 and 12% for retention time and peak area, respectively, whereas the interday repeatability values were lower than 0.6 and 14% for retention time and peak area, respectively. Analyte recoveries found were 86 and 93% for capsaicin and dihydrocapsaicin, respectively. The method developed has been applied to the identification and quantification of capsaicin and dihydrocapsaicin in fruit extracts from different Capsicum genotypes, and concentrations found ranged from 2 to 6639 mg kg(-1).

Capsaicin↗

Direct connection of supercritical fluid extraction and supercritical fluid chromatography as a rapid quantitative method for capsaicinoids in placentas of Capsicum.

The fruits of Capsicum annuum L. are used worldwide as chili peppers and in folk medicines. The pungent components of C. annuum, which are irritants, are called capsaicinoids (CAPS), and the most abundant components are capsaicin, dihydrocapsaicin, and nordihydrocapsaicin. To analyze CAPS in the placentas of Capsicum fruits rapidly and safely, we used a directly connected system of supercritical fluid extraction and supercritical fluid chromatography (SFE/SFC). As a column for SFE/SFC, only a silica-type column was found to be suitable. The CAPS contents in placentas of C. annuum cv. Jalapeno (hot type) and C. annuum cv. Shishitoh (less-hot type) determined by the SFE/SFC method agreed well with those in the range of 0-13.81 mg g(-1) fr. wt determined by the usual extraction-HPLC method. The SFE/SFC method has the advantages of no need for pretreatment and no (or minimal) need for organic solvents. We conclude that this method is useful as a rapid (20 min) and safe screening test for the pungency of various Capsicum fruits.

Capsaicin↗

Changes in phytochemical and antioxidant activity of selected pepper cultivars (Capsicum species) as influenced by maturity.

The effect of fruit maturation on changes in carotenoids, flavonoids, total soluble reducing equivalents, phenolic acids, ascorbic acid, and antioxidant activity (AOX) in different pepper types (Capsicum annuum, Capsicum frutescens, and Capsicum chinese) was determined. Generally, the concentration of these chemical constituents increased as the peppers reached maturity. Peppers contained high levels of L-ascorbic acid and carotenoids at maturity, contributing 124-338% of the RDA for vitamin C and 0.33-336 RE/100 g of provitamin A activity, respectively. Levels of phenolic acids, capxanthin, and zeaxanthin generally increased during maturation, whereas the level of lutein declined. Flavonoid concentrations varied greatly among the pepper types analyzed and were negatively correlated to AOX under the conditions of the beta-carotene-linoleic assay. Model systems were used to aid in understanding the relationship between flavonoids and AOX. Significant increases in AOX were observed in pepper juice models in response to increasing dilution factors and the presence of EDTA, indicating a pro-oxidant effect due to metal ions in the system. In vitro models demonstrated that increasing levels of flavonoids in combination with constant levels of caffeic and ascorbic acid gave a resultant AOX that was either additive of the two compounds or competitive in their ability to scavenge peroxyl radicals. The model systems were in good agreement with the chemical composition of the pepper cultivars and reflected the interactions affecting AOX. More research is needed to understand the complex interactions that occur among various antioxidants present in pepper extracts.

Antioxidants↗