Key message
CAP biosynthesis in the pericarp of chili pepper fruits occurs with an ambiguous boundary in the placental septum and pericarp.
Capsaicinoid (CAP) is a pungent ingredient of chili pepper ...fruits. Generally, CAP biosynthesis is limited to the placental septum of fruits, but it has been reported that its biosynthesis occurs even in the pericarp of some extremely pungent varieties, resulting in a substantial increase in total content. To examine the mechanism of CAP biosynthesis in the pericarp, comparative transcriptome analysis of a variety that produces CAP in the pericarp (MY) and a variety that does not (HB) was carried out. RNA-seq revealed that 2264 genes were differentially expressed in the MY pericarp compared with the HB pericarp. PCA analysis and GO enrichment analysis indicated that the MY pericarp has a gene expression profile more like placental septum than the HB pericarp. The gene expression of CAP biosynthesis-related genes in the MY pericarp changed coordinately with the placental septum during fruit development. In most
Capsicum
accessions including HB, the distribution of slender epidermal cells producing CAP was limited to the placental septum, and the morphological boundary between the placental septum and pericarp was clear. In some extremely pungent varieties such as MY, slender epidermal cells ranged from the placental septum to the pericarp region, and the pericarp was morphologically similar to the placental septum, such as the absence of large sub-epidermal cells and abundant spaces in the parenchymal tissue. Our data suggest that CAP biosynthesis in the pericarp occurred with an ambiguous boundary in the placental septum and pericarp. These findings contribute to further enhancement of CAP production in chili pepper fruits.
Introduction
Proton (1H) nuclear magnetic resonance (NMR) spectroscopy based analytical method for the quantification of capsaicin (major pungent principle of chili) has certain advantages including ...short data acquisition time and better structural authentication. Earlier NMR methods are associated with either of the bottlenecks such as low or lack of information on the sensitivity and scope for the quantification of total capsaicinoid.
Objective
To develop a sensitive 1H quantitative NMR (qNMR) technique for capsaicin and total capsaicinoid in dry chili and chili oleoresin and to demonstrate its applicability in a real sample set.
Method
A 1H qNMR method was developed using benzene as the internal standard for the quantification of capsaicin (terminal methyl signal) as well as total capsaicinoid (benzyl methylene signal) in dry chili and oleoresin and validated in terms of specificity, linearity, sensitivity, accuracy and precision.
Results
The developed 1H qNMR method was specific, sensitive (limit of detection 4.4 μg/mL and limit of quantitation 14.8 μg/mL), linear in the range 0.083–8.33 mg/mL of capsaicin, accurate and precise. The credibility of the developed method was showcased in the morpho‐chemical characterisation of commercially available 15 chili land races from northeast India. The analysis identified the land races with a wide range of capsaicin (trace to 1.49% in the dry fruit and trace to 6.21% in the oleoresin w/w) and oleoresin content (3.35–26.78% w/w).
Conclusion
The standardized 1H qNMR method facilitated the findings of chemical basis for the selection of chili land races from this region, capable of producing high‐yielding oleoresin with intended degree of pungency.
A standardized 1H quantitative NMR (qNMR)method was developed for the quantification of capsaicin and total capsaicinoid in dry chili and chili oleoresin. The developed method was sensitive with a limit of detection 4.4 μg/mL and limit of quantitation 14.8 μg/mL for capsaicin. The applicability of the developed method was shown through the morpho‐chemical characterisation of commercially available 15 chili landraces from northeast India. The study identified a wide variability in oleoresin and capsaicin content among the studied landraces.
The study aims to identify the morphological and physiological variation of the 4th generation of C. frutescens L. G1/01 and G7/01. The morphological data measured based on IPGRI, AVRDC and CATIE ...descriptors, and capsaicinoid content measured using spectrophotometry were used to build Jaccard similarity-based cladogram. The results showed that 40% of the G1/01 and 100% of the G7/01 plant height were categorized into very high category (> 85 cm). One hundred percent of the G1/01 and G7/01 were categorized into very early and late flowering, respectively. Eighty-five percent of the G1/01 and 100% of the G7/01 fruit length were categorized into normal (1-5cm). Ninety percent of the G1/01 and 56% of the G7/01 fruit diameters were categorized into normal (>0.5-1cm). Eighty-nine percent of G1/01 and 100% of G7/01 fruit weight were categorized into moderate (0.1-2g). Eighty percent of the G1/01 and 100% of the G7/01 number of seeds were categorized into moderate (>50-200 seeds) and small (<50 seeds), respectively. The capsaicinoid content of G1/01 and G7/01 was significantly lower than the original type. Mutant T39 (G7/01) had the highest capsaicinoid content (19.35mg/g). In the dendrogram, genotypes G1/01 and G7/01 were separated based on the character of the node color, petal color, peel surface, leaf color, and flowering time. G1/01 formed five clusters while G7/01 formed two clusters. G1/01 is more varied than G7/01.
In Capsicum, loss of function mutation of acyltransferase (Pun1), putative aminotransferase (pAMT), putative ketoacyl-ACP reductase (CaKR1), and R2R3-MYB transcription factor (CaMYB31) have been ...reported to be the genetic causes of non-pungency. In the present study, 245 C. chinense accessions were initially screened for non-pungency attributes. Six candidates with identification numbers, No. 3327, No. 3356, No. 3529, No. 4026, No. 4028, and No. 4034 were selected by tasting test, and the non-pungency attribute was confirmed by high-performance liquid chromatographic analysis. Expression and sequence analysis inferred that the non-pungency of No. 3529 was due to the non-expression of Pun1. Analysis of pAMT confirmed that No. 3356 (pamt5) and No. 4034 (pamt9) had loss of function mutations. Because the non-pungency of No. 3327, No. 4026, and No. 4028 did not seem to be caused by mutation of either Pun1 or pAMT, the CaKR1 mutation was further examined using a polymerase chain reaction-based, co-dominant marker. Genotyping clarified that No. 3327, No. 4026, and No. 4028 had the same mutated CaKR1 allele as non-pungent No. 3341. Moreover, a crossing test with a pungent Habanero and No. 3341 clearly revealed that the non-pungency in No. 3327, No. 4026, and No. 4028 was a result of a loss of function mutation of CaKR1. Our previous and present studies have shown that non-pungent cultivars of C. chinense possessing pamt are widely distributed in Central America, South America and the West Indies (Caribbean), while non-pungent cultivars possessing Cakr1 originate from Bolivia and Peru. Some artificial selection may have occurred that was based on a preference for non-pungent peppers in the local region of origin.
Capsicum species produce fruits that synthesize and accumulate unique hot compounds known as capsaicinoids in placental tissues. The capsaicinoid biosynthetic pathway has been established, but the ...enzymes and genes participating in this process have not been extensively studied or characterized. Capsaicinoids are synthesized through the convergence of two biosynthetic pathways: the phenylpropanoid and the branched-chain fatty acid pathways, which provide the precursors phenylalanine, and valine or leucine, respectively. Capsaicinoid biosynthesis and accumulation is a genetically determined trait in chili pepper fruits as different cultivars or genotypes exhibit differences in pungency; furthermore, this characteristic is also developmentally and environmentally regulated. The establishment of cDNA libraries and comparative gene expression studies in pungent and non-pungent chili pepper fruits has identified candidate genes possibly involved in capsaicinoid biosynthesis. Genetic and molecular approaches have also contributed to the knowledge of this biosynthetic pathway; however, more studies are necessary for a better understanding of the regulatory process that accounts for different accumulation levels of capsaicinoids in chili pepper fruits.
ABSTRAK Pertimbangan dalam pemilihan metode ekstraksi untuk mendapatkan senyawa bioaktif dari bahan tanaman diantaranya adalah efisiensi waktu dan jumlah pelarut yang digunakan. Studi terdahulu ...menunjukan ekstraksi bahan aktif dengan gelombang mikro mempercepat waktu ekstraksindan menghemat penggunaan pelarut dibandingkan metode konvensional, tetapi waktu ekstraksibyang terlalu singkat dan penggunaan pelarut yang terlalu minim membuat hasil ekstraksi tidaknoptimal. Penelitian ini untuk mengetahui pengaruh rasio bahan:pelarut dan lama ekstraksi dengan gelombang mikro terhadap kandungan fitokimia dan aktivitas antioksidan dari ekstrak cabai rawit. Penelitian dilakukan menggunakan Rancangan Acak Kelompok 2 faktor yaitu rasio bahan:pelarut dengan variasi 1:5; 1:10; dan 1:15, serta lama ekstraksi (5, 10, dan 15 menit). Berdasarkan hasil pengujian, rasio bahan:pelarut 1:10 memberikan hasil terbaik untuk kadar total capsaicinoid, total fenol, total flavonoid, dan aktivitas antioksidan dengan nilai rata-rata sebesar 138.06 ± 6.35 mg CE/g BK ekstrak, 33.75 ± 1.75 mg GAE/g BK ekstrak, 205.41 ± 26.13 mg QE/g BK ekstrak, dan 447.96 ±25.63 μg/ml. Lama ekstraksi untuk menghasilkan nilai total capsaicinoid dan total fenol paling tinggi adalah 5 menit, sedangkan untuk total flavonoid pada waktu 10 dan 15 menit. Nilai IC50 memiliki korelasi yang linear dengan total capsaicinoid, total fenol, dan total flavonoid. Pemilihan perlakuan terbaik dengan metode multiple atribut menunjukan rasio bahan:pelarut 1:10 dan lama ekstraksi 10 menit merupakan kombinasi perlakuan paling optimal untuk ekstraksi senyawa fenol menggunakan Microwave Assisted Extraction ABSTRACT The consideration of extraction methods to obtain bioactive compounds of plant origin were the time efficiency and the amount of extraction solvent. Previous studies showed that microwave assisted extraction of active ingredients decrease the extraction time and minimize the solvent compared to conventional methods, but short extraction time and less solvents make extraction less optimal. The aim of this study to determine the effect of materials:solvent and extraction time of microwave assisted extraction to phytochemical content and antioxidant activity of the cayenne pepper extract. The study was conducted using a Randomized Block Design 2 factors of the ratio of material:solvent (1:5; 1:10; and 1:15) and extraction time (5, 10, and 15 min). The results showed that material:solvent ratio of 1:10 gave the best value for total capsaicinoid content, total phenol, total flavonoid and IC50 with average value of 138.06 ± 6.35 mg CE/g DW, 33.75 ± 1.75 mg GAE/g DW extract, 205.41 ± 26.13 mg QE/g DW extract, and 447.96 ± 25.63 μg/ml respectively. The optimal extraction time to generate total capsaicinoid and total phenol content was 5 minutes, while the optimal extraction time for total flavonoid was 10 and 15 minutes. IC50 in this study had a linear correlation with total capsaicinoid, total phenol, and total flavonoid. The best treatment with multiple attribute method showed the materials:solvent ratio of 1:10 and extraction time of 10 minutes was the most optimal treatment combination for extraction of phenol compound by microwave assisted extraction
Capsaicinoid content, among other factors, affects the perception of spiciness of commercial kimchi. Here, we investigated whether the physicochemical properties of kimchi affect the spicy taste of ...capsaicinoids perceived by the tasting. High-performance liquid chromatography (HPLC) was used to evaluate the capsaicinoid content (mg/kg) of thirteen types of commercial kimchi. The physicochemical properties such as pH, titratable acidity, salinity, free sugar content, and free amino acid content were evaluated, and the spicy strength grade was determined by selected panel to analyze the correlation between these properties. Panels were trained for 48 h prior to actual evaluation by panel leaders trained for over 1000 h according to the Spectrum
TM
method. Partial correlation analysis was performed to examine other candidate parameters that interfere with the sensory evaluation of spiciness and capsaicinoid content. To express the specific variance after eliminating the effects of other variables, partial correlations were used to estimate the relationships between two variables. We observed a strong correlation between spiciness intensity ratings and capsaicinoid content, with a Pearson’s correlation coefficient of 0.78 at
p
≤ 0.001. However, other specific variables may have influenced the relationship between spiciness intensity and total capsaicinoid content. Partial correlation analysis indicated that the free sugar content most strongly affected the relationship between spiciness intensity and capsaicinoid content, showing the largest first-order partial correlation coefficient (r
xy/z
: 0.091,
p
≤ 0.01).
Capsaicinoid nonivamide (PAVA) and rosuvastatin (RSV) have been shown to exert antioxidant and anti‐obesity effects in various animal models, but it is unknown whether their combination would be an ...effective treatment for obesity‐related endothelial dysfunction. This study aimed to investigate the mechanism of PAVA in synergy with RSV. Male Sprague–Dawley rats were given a high‐fat diet (HFD) or normal diet during a 20‐week period. At 16 weeks, rats in each diet group were divided into subgroups. Normal diet rats were divided into Normal diet control, Normal diet with PAVA, and Normal diet with RSV groups. HFD rats were subdivided into HFD control, HFD with PAVA, HFD with RSV, and HFD with PAVA + RSV groups and evaluated for metabolic parameters, blood pressure, aortic function, and histological change of the aorta in rats. Our results showed the combined therapy had a significantly greater effect than the monotherapy in all measured parameters; this was indicated by improvement in insulin sensitivity and aortic function, decreased blood pressure, lower oxidative stress, and prevention of vascular damage. The synergistic effect of the PAVA and RSV can protect HFD‐induced obesity‐related endothelial dysfunction, suggesting that the combination of PAVA and RSV could be an effective alternative treatment for obesity‐related complications in patients with cardiovascular disease.
The capsaicinoid nonivamide (PAVA) and rosuvastatin (RSV) have been shown to exert antioxidant and anti‐obesity effects in various animal models. The present study reveals that the synergistic effect of the capsaicinoid nonivamide and rosuvastatin can protect high‐fat diet‐induced obesity‐related endothelial dysfunction by improving in insulin sensitivity and aortal function as well as decreasing blood pressure, lowering oxidative stress, and preventing of vascular damage. Its effect could be beneficial for the health of patients with cardiovascular disease.
Transient receptor potential vanilloid 1 (TRPV1) activation by capsaicin binding increased intracellular calcium influx and stimulated adipocyte-to-adipocyte communication, leading to lipolysis. ...Generally, enhancement of π-stacking capabilities improves certain binding interactions. Notably, nitroarenes exhibit strong binding interactions with aromatic amino acid side chains in proteins. New capsaicinoid analogs were designed by substitution of the OCH3 group with a nitrogen dioxide (NO2) group on the vanillyl ring to investigate how π-stacking interactions in capsaicinoid analogs contribute to lipolysis. Capsaicinoid analogs, nitro capsaicin (5), and nitro dihydrocapsaicin (6) were prepared in moderate yields via coupling of a nitroaromatic amine salt and fatty acids. Oil Red O staining and triglyceride assays with 10 µM loading of capsaicin (CAP), dihydrocapsaicin (DHC), 5, and 6 were performed to investigate their effect on lipolysis in 3T3-L1 adipocytes. Both assay results indicated that 5 and 6 decreased lipid accumulation by 13.6% and 14.7%, respectively, and significantly reduced triglyceride content by 26.9% and 28.4%, respectively, in comparison with the control experiment. Furthermore, the decrease in triglyceride content observed in response to nitroarene capsaicinoid analogs was approximately 2-folds higher than that of CAP and DHC. These results arose from the NO2 group augmented π-π stacking with Tyr511 and the attractive charge interaction with Glu570 affecting binding interactions with TRPV1 receptors.
•The time-intensity parameter Imax depends on the complexity of the matrix.•Pungency perception is strongly influenced by the starch content in food.•Results obtained from model food matrices can be ...applied to convenience products.•The HPLC analysis and the sensory evaluation (Imax) correlate with an R2 of 0.8029.
The influence of food ingredients (water, starch, fat, and sugar), individually and in combination, on the sensory perception of the pungency of capsaicin was studied in model food systems using the time-intensity method. Furthermore, the transferability of the results obtained using model food matrices to complex matrices, such as convenience salsas, was investigated. Significant differences between the samples (p<0.05) were examined by analysis of variance (ANOVA) followed by the Tukey-HSD post hoc test. A highly complex matrix and a high starch content reduced the perception of pungency in both the model food matrices and the convenience salsa products. Therefore, the time-related maximum intensity of pungency (Imax) was affected by other factors in addition to the capsaicin concentration.
The total capsaicinoid content in the salsas was quantified by high-performance liquid chromatography–mass spectrometry (HPLC–MS) and correlated to the results of the sensory evaluation. The best correlation (R2=0.8029) was observed for the total capsaicinoid content and the maximum intensity of pungency (Imax) in comparison to the duration of the total pungency perception TDur (R2=0.5176) and the area under the curve (R2=0.6898). The calculated regression models serve as an example of how empirical calculations can be generated and used in a specific context.
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