Peppermint Essential Oil and Its Major Volatiles as Protective Agents against Soft Rot Caused by Fusarium sambucinum in Cera Pepper (Capsicum pubescens).

Cera pepper (Capsicum pubescens) is an exotic fruit considered as a rich source of nutraceuticals with known benefits for human health and also an economic resource for local producers in Mexico. The present investigation reports on the in vitro and in situ antifungal activity of the essential oil from Mentha piperita and its two major volatiles (menthol and menthone) against Fusarium sambucinum, which is a causal agent of soft rot in cera pepper. The application of these components in pepper fruits previously infected with F. sambucinum caused a significant delay (p<0.05) in the emergence of soft rot symptoms. This effect was reflected in the maintenance of pH and fruit firmness during a period of 10 days. The nutrimental content of the fruits (protein, fiber, fat and other proximate parameters) was conserved in the same period of time. The nutraceutical content of these fruits was estimated by the quantification of seven carotenoids (violaxanthin, cis-violaxanthin, luteoxanthin, antheraxanthin, lutein, zeaxanthin and ß-carotene), ascorbic acid and capsaicinoids (capsaicin and dihydrocapsaicin). According to our results, the essential oil from M. Piperita and its major volatiles exerted a preservative effect on these metabolites. Our findings demonstrated that the essential oil of M. Piperita and its major volatiles represent an ecological alternative for the control of fusariosis caused by F. sambucinum in cera peppers under postharvest conditions.

Correlation of Chillies Capsaicinoids Contents with their Cytotoxic Effects against Hepatocarcinoma Cells.

BACKGROUND AND OBJECTIVE: Chillies are delicious spices that are used extensively. Capsaicinoids, the major constituents of chillies with reported anti-cancer effects, have been determined with non-specific colorimetric methods. A rapid and reproducible method for extraction and quantification of the major chillies capsaicinoids; capsaicin, dihydrocapsaicin (DHC) and nordihydrocapsaicin (n-DHC), was reported, moreover study of their cytotoxic activity. MATERIALS AND METHODS: This study has covered the extraction of capsaicinoids from red and green-colored chillies followed by their quantification using HPLC-UV method after validation. Furthermore, the correlation of capsaicinoids contents with their in vitro hepatocarcinoma cytotoxicity was represented by Pearson's correlation coefficient. RESULTS: Capsaicinoids contents are ranged from 1219.88-15098.67 ng mg-1 of Dried Extract (DE). Capsaicin exhibits the lowest IC50 when compared to doxorubicin (9.201±0.91 and 16.1±0.82 µg mL-1, respectively). The exhibited activities of methanol extracts of red and green-colored chillies (IC50 = 20.21±1.72 and 16.02±0.69 µg mL-1, respectively) may attribute to their excessive contents of capsaicinoids (6975.42 and 15098.67 ng mg-1 DE, respectively). Capsaicin and n-DHC contents have a negative correlation with cytotoxic activity. CONCLUSION: Green-colored chillies were found to be more cytotoxic in comparison with red-colored chillies that may be relative to their high content of capsaicinoids. The present investigation suggests that capsaicinoids contents correlate with cytotoxic activity.

Determination of capsaicinoids by magnetic solid phase extraction coupled with UPLC-MS/MS for screening of gutter oil.

Capsaicinoids, including capsaicin, dihydrocapsaicin and N-vanillylnonanamide have been identified as characteristic markers for gutter oil. In this study, the nanocomposite of graphene oxide-Fe3O4 (GO-Fe3O4) was synthesized by a co-precipitation method, and applied to magnetic solid phase extraction (MSPE) process. The capsaicin, dihydrocapsaicin and N-vanillylnonanamide extracted in gutter oil were analyzed by ultrahigh performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Under the optimized pretreatment conditions, the limit of detection (LOD) and limit of quantification (LOQ) are 0.15 and 0.4 µg L-1, respectively. The linear range is 0.4-200 µg L-1, and the correlation coefficients (R2) are over 0.996 for all three compounds. The recoveries range from 81.2% to 103.3% for soybean oil and 71.2% to 84.3% for rapeseed oil. The values of matrix effect range in 87.6%-105.5% and 83.4%-110.6% and the intra-day RSDs range from 2.8%-4.6% to 4.8%-6.1% for soybean oil and rapeseed oil, respectively. The adsorbent can be reused for multiple times with no appreciable loss of adsorption capability. The present methodology has high potential for screening of gutter oil in food samples, thus having importance for food safety monitoring.

Capsaicinoid nonivamide improves nonalcoholic fatty liver disease in rats fed a high-fat diet.

Nonalcoholic fatty liver disease (NAFLD) is a chronic disease that causes morbidity associated with metabolic syndrome. NAFLD is a worldwide problem and represents a major cause of liver injury, which can lead to liver cell death. We investigated the effects of nonivamide (pelargonic acid vanillylamide, PAVA; 1 mg/kg) and rosuvastatin (RSV; 10 mg/kg) on hepatic steatosis induced by a high-fat diet (HFD). Male Sprague-Dawley rats were fed a HFD for 16 weeks then received PAVA or RSV for 4 additional weeks. We examined the metabolic parameters, function, fat content, histological alterations, reactive oxygen species production, and apoptotic cell death of the liver, in addition to the expression of the following important molecules: transient receptor potential cation channel subfamily V member 1 (TRPV1) phosphorylation of sterol regulatory element binding protein (pSREBP-1c/SREBP-1c), total and membrane glucose transporter 2 (GLUT2), 4-hydroxynonenal (4-HNE), and cleaved caspase-3. HFD-induced hepatic steatosis was associated with significantly increased morphological disorganization, injury markers, oxidative stress, lipid peroxidation, and apoptosis. However, metabolic dysfunction and hepatic injury were reduced by RSV and PAVA treatment. PAVA regulated lipid deposition, improved insulin resistance, and decreased oxidative stress and apoptotic cell death. Therefore, PAVA represents a promising therapeutic approach for treating metabolic disorders in patients with NAFLD.

Extraction and Purification of Capsaicin from Capsicum Oleoresin Using a Combination of Tunable Aqueous Polymer-Phase Impregnated Resin (TAPPIR) Extraction and Chromatography Technology.

Capsaicin, which mainly comes from pepper, exhibits anticancer, antioxidant, and anti-obesity properties. This work aims to construct a comprehensive technology for the extraction and purification of capsaicin from capsicum oleoresin. The tunable aqueous polymer phase impregnated HZ816 resins were selected in extraction step. In the extraction process, 3 g of impregnated HZ816 macroporous resin was employed per system. The results showed that a higher molecular weight of Polyethylene glycol (PEG) and 1-ethyl-3-methyl imidazolium acetate ([Emim] [OAc]) are more beneficial to the improvement of the yield of capsaicin. Screening experiment using fractional factorial designs indicated that the amount of sample loading, pH, and concentration of [Emim] [OAc] and PEG 6000 significantly affect the yield of capsaicin. Mathematical models of capsaicin yield in tunable aqueous polymer-phase impregnated resins were established and optimum condition was obtained using response surface methodology. The optimum impregnated phase was the polymer phase of an aqueous two-phase system which contained 18.5% (w/w) PEG6000, 15% (w/w) sodium citrate, and 10% (w/w) [Emim] [OAc] at pH 6.5. Under the optimal conditions, the yield of capsaicin reached 95.82% when the extraction system contains 0.25 g capsicum oleoresin. Ultimately, capsaicinoids extract was purified by reverse-phase resin (SKP-10-4300) chromatographic column. The capsaicin recovery and purity achieved 85% and 92%, respectively.

Isolation of soluble epoxide hydrolase inhibitor of capsaicin analogs from Capsicum chinense Jacq. cv. Habanero.

Capsaicin (1) and dihydrocapsaicin (2) were isolated from the aerial parts of Capsicum chinense Jacq. cv. Habanero. In in vitro studies, the exhibited potent inhibitory activity against soluble epoxide hydrolase (sEH), with IC50 values of 5.6 ±â€¯1.2 and 7.3 ±â€¯0.7 µM. Enzyme kinetics suggested that the two compounds (1 and 2) were competitive inhibitors with Ki values of 2.5 ±â€¯1.0 and 4.7 ±â€¯2.3 µM, respectively. Molecular docking and molecular dynamic studies showed that capsaicin analogs (1 and 2) bound strongly with Asp335, Tyr383, and Tyr466 residues in the active site. These results suggest that the two inhibitors (1 and 2) represent potentially therapeutic inhibitors of sEH.

Green approach to the extraction of major capsaicinoids from habanero pepper using near-infrared, microwave, ultrasound and Soxhlet methods, a comparative study.

A simple method for the extraction of two major capsaicinoids from habanero peppers, using near-infrared irradiation, microwave irradiation or ultrasound as the energy source and ethanol as the solvent, was compared with Soxhlet extraction. The extraction processes were monitored by gas chromatography/mass spectrometry. The new processes offer better overall yields and a higher ratio of capsaicin to dihydrocapsaicin than Soxhlet extraction. The physical effect of the different energy sources on the peppers was determined using scanning electron microscopy. Extraction of capsaicin and dihydrocapsaicin using near-infrared irradiation, which has not been previously reported, was shown to be a simple and efficient alternative extraction procedure.

Hybrid electrochemical sensor platform for capsaicin determination using coarsely stepped cyclic squarewave voltammetry.

A small, standalone electrochemical hybrid sensor platform, combining flexible electronics and screen-printed electrodes, is demonstrated in the determination of capsaicin through adsorptive stripping voltammetry. The sensing scheme was simplified to be compatible with a low-cost device. The simplification involved eliminating the need for additional modification of the electrode and employing a coarsely stepped squarewave voltammetry, a technique which is applicable with less sophisticated instrumentation. This architecture was found to be suitable for concentrations up to at least 5000 µM with a detection limit of 1.98 µM. The screen-printed carbon graphite electrodes were made reusable through an ethanol rinsing protocol. The effect of ethanol/buffer volumetric ratio in the test sample was shown to greatly influence the analytical data, and a fixed 10% (v/v) was chosen as a compromise between signal-to-noise ratio and not exceeding the solubility limit of the desired upper range.

Quantification of capsaicinoids in chillies by solid-phase extraction coupled with voltammetry.

Capsaicinoids were extracted from a range of spices and chillies using methanol, prior to concentrating the compounds using solid-phase extraction cartridges and water/methanol (50:50% v/v) as the solvent, followed by elution with acetonitrile. The primary extraction procedure, involving only sonication of the spices in methanol, gave results comparable to a procedure that used a combination of sonication, stirring and centrifuging. The voltammetric quantification of the capsaicinoids, at approximately +0.5 V vs. ferrocene0/+ that were transferred from methanol/water into acetonitrile/water via solid phase extraction, was carried out in microcentrifuge tubes. Linear calibration curves for voltammetry measurements were obtained from low ppm up to at least 1400 ppm of capsaicinoids, with concentrations being detected in the different source extracts (paprika, tabasco sauce, cayenne pepper, and fresh chillies) from approximately 17 to 430 ppm, which corresponded to values of between approximately 130 and 4000 ppm, respectively, present in the original samples.

Capsaicin and Piperine Can Overcome Multidrug Resistance in Cancer Cells to Doxorubicin.

BACKGROUND: Multidrug resistance (MDR) can develop in cancer cells after treatment with anticancer drugs, mainly due to the overexpression of the ATP-binding cassette (ABC) transporters. We analyzed the ability of two pungent-tasting alkaloids-capsaicin and piperine from Capsicum frutescens and Piper nigrum, respectively-to reverse multidrug resistance in the cancer cell lines Caco-2 and CEM/ADR 5000, which overexpress P-glycoprotein (P-gp) and other ABC transporters. METHODS: The MTT assay was first used to determine the cytotoxicity of doxorubicin, the alkaloids, and digitonin alone, and then their combinations. Furthermore, rhodamine (Rho) 123 and calcein-AM were used to detect the effects of alkaloids on the activity of P-gp. RESULTS: Capsaicin and piperine synergistically enhanced the cytotoxicity of doxorubicin in Caco-2 and CEM/ADR 5000 cells. Furthermore, capsaicin and piperine increased the intracellular accumulation of the fluorescent P-glycoprotein (P-gp) substrates rhodamine and calcein and inhibited their efflux from the MDR cell lines. CONCLUSION: Our study has demonstrated that capsaicin and piperine are P-gp substrates and have potential chemosensitizing activity, which might be interesting for the development of novel modulators of multidrug resistance.

Preparative isolation and purification of capsaicin and dihydrocapsaicin from Capsici Fructus using supercritical fluid extraction combined with high speed countercurrent chromatography.

BACKGROUND: Supercritical fluid extraction with CO2 (SFE-CO2 ) was utilized for extraction of capsaicin (CA) and dihydrocapsaicin (DHCA) from Capsici Fructus, and then a two-step enrichment method for separating capsaicinoids from SFE-CO2 extracts was developed. The process involved extraction with aqueous methanol and crystallization by alkali extraction and acid precipitation. Finally, a consecutive high-speed countercurrent chromatography (HSCCC) separation method was successfully applied in the purification of CA and DHCA from capsaicinoid crystal. RESULTS: The extraction pressure, extraction temperature and volume of co-solvent were optimized at 33 MPa, 41 °C and 75 mL, respectively, using response surface methodology; the extraction rates of CA and DHCA were about 93.18% and 93.49%, respectively. 407.43 mg capsaicinoid crystal was isolated from the SFE-CO2 extracts obtained from 100 g capsicum powder by the two-step enrichment method. About 506 mg and 184 mg CA and DHCA with purities up to 98.31% and 96.68%, respectively, were obtained from 1 g capsaicinoid crystal in one HSCCC of three consecutive sample loadings without exchanging any solvent system. CONCLUSIONS: This method comprising SFE-CO2 , a two-step enrichment and HSCCC was efficient, powerful and practical for the large-scale preparation of CA and DHCA from Capsici Fructus with high purity and high yield. © 2017 Society of Chemical Industry.

Rapid determination of capsaicinoids by colorimetric method.

Capsaicinoids, the pungent component of chili peppers, are generally analyzed by precise analytical techniques, such as gas chromatography and high-performance liquid chromatography (HPLC), but these are not practical for the mass analyses of samples. To analyze mass samples rapidly, a colorimetric method was suggested. In this work, pigments and capsaicinoids were efficiently separated from chili pepper extract by sequential solid-liquid extraction and liquid-liquid extraction in test tubes followed by a colorimetric analysis on the capsaicinoids by a selective chromogenic reaction with Gibbs reagent (2,6-dichloroquinone-4-chloroimide). In the comparison of the capsaicinoid content by the colorimetric method and HPLC using acetone extracts of fresh pepper and dry red pepper as samples, R2 was 0.9973 and 0.9816, respectively, which shows a high linear correlation. In addition, a minimum of 1 µg/mL capsaicinoids can be detected and it was therefore determined that the method can efficiently analyze a great quantity of samples in a short time.

Extraction and purification of capsaicin from capsicum oleoresin using an aqueous two-phase system combined with chromatography.

Capsaicin was extracted from capsicum oleoresin using an aqueous two-phase system (ATPS) composed of an ethylene oxide-propylene oxide (EOPO) copolymer, salt and ethanol. Capsaicin was concentrated in the top polymer-rich phase. To determine the optimal conditions, the partitioning of capsaicin in the ATPS was investigated, considering a single-factor experiment including the salt concentration, polymer concentration, buffer pH, ethanol concentration, sample loading and extraction duration. Response surface methodology was applied to investigate the effects of the polymer concentration, buffer pH and sample loading on capsaicin partitioning. A capsaicin yield of 95.5% was obtained using the optimal extraction system, which consisted of 16.3% UCON 50-HB-5100/10% K2HPO4/1% ethanol, a buffer pH of 4.35 and 0.24g of capsicum oleoresin. Capsaicin was purified from the capsaicinoid extract using a two-step macroporous adsorption resin (MAR) method. After purification using non-polar MAR ADS-17, the recovery and purity of capsaicin were 83.7% and 50.3%, respectively. After purification using weakly polar MAR AB-8, the recovery and purity of capsaicin were 88.0% and 85.1%, respectively.

Ontogenetic Variation of Individual and Total Capsaicinoids in Malagueta Peppers (Capsicum frutescens) during Fruit Maturation.

The ontogenetic variation of total and individual capsaicinoids (nordihydrocapsaicin (n-DHC), capsaicin (C), dihydrocapsaicin (DHC), homocapsaicin (h-C) and homodihydrocapsaicin (h-DHC)) present in Malagueta pepper (Capsicum frutescens) during fruit ripening has been studied. Malagueta peppers were grown in a greenhouse under controlled temperature and humidity conditions. Capsaicinoids were extracted using ultrasound-assisted extraction (UAE) and the extracts were analyzed by ultra-performance liquid chromatography (UHPLC) with fluorescence detection. A significant increase in the total content of capsaicinoids was observed in the early days (between 12 and 33). Between day 33 and 40 there was a slight reduction in the total capsaicinoid content (3.3% decrease). C was the major capsaicinoid, followed by DHC, n-DHC, h-C and h-DHC. By considering the evolution of standardized values of the capsaicinoids it was verified that n-DHC, DHC and h-DHC (dihydrocapsaicin-like capsaicinoids) present a similar behavior pattern, while h-C and C (capsaicin-like capsaicinoids) show different evolution patterns.

Therapeutic Strategies of Plant-derived Compounds for Diabetes Via Regulation of Monocyte Chemoattractant Protein-1.

BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) is a member of the CC chemokine family that plays a key role in the inflammatory process. It has been broadly studied in the aspect of its role in obesity and diabetes related diseases. MCP-1 causes the infiltration of macrophages into obese adipose tissue via binding to the CCR2 receptor and is involved in the development of insulin resistance. METHODS: We reviewed the available literature regarding the importance of plant metabolites that regulate MCP-1 activity and are used in the treatment of diabetic disorders. The characteristics of screened papers were described and the important findings were included in this review. RESULTS: This mini-review provides a summary of functions and therapeutic strategies of this chemokine, with a special focus on plant-derived compounds that possess a putative antidiabetic function via a mechanism of MCP-1 interaction. The highlights of this review include the roles of MCP-1 in development of diabetes, the evaluation of plant metabolites that specifically or non-specifically inhibit MCP-1 overproduction, and the molecular mechanisms of this activity. Among these metabolites, we particularly focused on phenolic acids and their derivatives, flavonoids, stilbenes, anthocyanins, capsaicin, alkaloids, plant sterols, terpenes, saponins, unsaturated fatty acids and plant-derived extracts. CONCLUSION: Regarding the increasing number of diabetic patients yearly, the recent progress in the putative therapies needs to be summarized. This article underlines the significance and involvement of the chemokine MCP-1 in the development of obesity, type 2 diabetes, and diabetic complications, with an emphasis on the role of plant metabolites in the regulation of this chemokine and thus the role in the prevention or therapy of diabetes. We suggest that MCP-1 might be a molecular marker of type 2 diabetes.

A simple analytical platform based on thin-layer chromatography coupled with paper-based analytical device for determination of total capsaicinoids in chilli samples.

A new analytical platform based on the use of thin-layer chromatography (TLC) coupled with paper-based analytical device (PAD) was developed for the determination of total capsaicinoids in chilli samples. This newly developed TLC-PAD is simple and low-cost without any requirement of special instrument or skillful person. The analysis consisted of two steps, i.e., extraction of capsaicinoids from chilli samples by using ethanol as solvent and separation of capsaicinoids by thin-layer chromatography (TLC) and elution of capsaicinoids from the TLC plate with in situ colorimetric detection of capsaicinoids on the PAD. For colorimetric detection, Folin-Ciocalteu reagent was used to detect phenolic functional group of capsaicinoids yielding the blue color. The blue color on the PAD was imaged by a scanner followed by evaluation of its grayscale intensity value by ImageJ program. This newly developed TLC-PAD method provided a linear range from 50 to 1000mgL-1 capsaicinoids with the limit of detection as low as 50mgL-1 capsaicinoids. The proposed method was applied to determine capsaicinoids in dried chilli and seasoning powder samples and the results were in good agreement with those obtained by HPLC method.

Capsaicin: Current Understanding of Its Mechanisms and Therapy of Pain and Other Pre-Clinical and Clinical Uses.

In this review, we discuss the importance of capsaicin to the current understanding of neuronal modulation of pain and explore the mechanisms of capsaicin-induced pain. We will focus on the analgesic effects of capsaicin and its clinical applicability in treating pain. Furthermore, we will draw attention to the rationale for other clinical therapeutic uses and implications of capsaicin in diseases such as obesity, diabetes, cardiovascular conditions, cancer, airway diseases, itch, gastric, and urological disorders.

The pepper's natural ingredient capsaicin induces autophagy blockage in prostate cancer cells.

Capsaicin, the pungent ingredient of red hot chili peepers, has been shown to have anti-cancer activities in several cancer cells, including prostate cancer. Several molecular mechanisms have been proposed on its chemopreventive action, including ceramide accumulation, endoplasmic reticulum stress induction and NFκB inhibition. However, the precise mechanisms by which capsaicin exerts its anti-proliferative effect in prostate cancer cells remain questionable. Herein, we have tested the involvement of autophagy on the capsaicin mechanism of action on prostate cancer LNCaP and PC-3 cells.The results showed that capsaicin induced prostate cancer cell death in a time- and concentration-dependent manner, increased the levels of microtubule-associated protein light chain 3-II (LC3-II, a marker of autophagy) and the accumulation of the cargo protein p62 suggesting an autophagy blockage. Moreover, confocal microscopy revealed that capsaicin treatment increased lysosomes which co-localized with LC3 positive vesicles in a similar extent to that produced by the lysosomal protease inhibitors E64 and pepstatin pointing to an autophagolysosomes breakdown inhibition. Furthermore, we found that capsaicin triggered ROS generation in cells, while the levels of ROS decreased with N-acetyl-cysteine (NAC), a ROS scavenger. Co-treatment of cells with NAC and capsaicin abrogated the effects of capsaicin on autophagy and cell death. Normal prostate PNT2 and RWPE-1 cells were more resistant to capsaicin-induced cytotoxicity and did not accumulate p62 protein.Taken together, these results suggest that ROS-mediated capsaicin-induced autophagy blockage contributes to antiproliferation in prostate cancer cells, which provides new insights into the anticancer molecular mechanism of capsaicin.