Exudate and Propolis from Escallonia pulverulenta: Phytochemical Characterization and Antibacterial Activity.

Propolis is a complex mixture formed from exudates that bees collect from plants and then mix with beeswax and their own salivary enzymes. Chilean propolis is characterized by the presence of phenolic compounds, which are considered responsible for the biological activities. The endemic species Escallonia pulverulenta (Ruiz and Pav.) Pers. [Escalloniaceae] is a recognized source of exudate to produce propolis. This study reports for the first time the chemical profile and antibacterial activity of E. pulverulenta exudate and leaves, as well as two samples of Chilean propolis. Palynological and morphological analysis showed the presence of E. pulverulenta as one of the main species in the propolis samples. UPLC-MS/MS analyses enabled the identification of phenolic acids in the leaves and in the propolis. Conversely, flavonoids are mainly present in exudates and propolis. Quercetin is the most abundant flavonol in the exudate, with similar concentrations in the propolis samples. Nevertheless, the main compound present in both samples of propolis was the flavanone pinocembrin. The antibacterial results obtained for exudate and propolis have shown a similar behavior, especially in the inhibition of Streptococcus pyogenes. These results show the importance of the exudates collected by the bees in the chemical composition and antibacterial capacity of propolis.

Biological capacities of seven Chilean ferns.

Sticherus squamulosus, Hymenophyllum dentatum, Blechnum microphyllum, Polypodium Feuillei, Blechnum chilense, Lophosoria quadripinnata, and Lissopimpla excelsa are native fern species found in southern Chile and are often used in traditional medicine. However, their bioactive properties have not been confirmed. In this study, ethanolic extracts of ferns exhibited significant antibacterial capacity against human pathogens. H. dentatum and S. squamulosus were selected for further analysis owing to their antimicrobial and antioxidant capacities. The entire H. dentatum extract exhibited fungistatic and fungicidal capacity on Botrytis cinerea and Penicillium digitatum, with minimum inhibitory concentration values of 1.56 mg/mL and 6.25 mg/mL and minimum fungicidal concentration values of 6.25 mg/mL and 12.5 mg/mL, respectively. The S. squamulosus extract presented the highest rutin content (222.5 µg/g fern). In addition, the ethanolic extract of H. dentatum inhibited 91% of DPPH radicals. Therefore, both types of ferns can be used in the food industry.

Phenolics from Chilean Bee Bread Exhibit Antioxidant and Antibacterial Properties: The First Prospective Study.

Bee bread (BB) is a beehive product generated upon fermentation of pollen combined with flower nectar and glandular secretions. The potential application of BB is related to its nutritional and functional components, including phenolic compounds. This is the first prospective study on palynological parameters, phenolics, antioxidant, and antibacterial activity of Chilean bee bread in vitro. The tested material exhibited high levels of phenolics (1340±186 mg GAE/100 g BB) and showed antioxidant capacity as determined by the FRAP (51±2 µmol Trolox equivalent/g BB) and ORAC-FL (643±64 µmol Trolox equivalent/g BB) and antibacterial activity against Streptococcus pyogenes. Furthermore, the phenolic acids and flavonoids was determined using liquid chromatography-mass spectrometry, and the concentration was determined using liquid chromatography with diode array detection. Kaempferol, quercetin, ferulic acid, and rutin were the main phenolics found. This study demonstrates the bioactive potential of Chilean BB and supports the evidence that this bee product is a promising source of antioxidants and antimicrobial compounds.

Floral Pollen Bioactive Properties and Their Synergy in Honeybee Pollen.

Honeybee pollen (HBP) is a mixture of floral pollen collected by honeybees near the hive. It is characterized by a composition rich in phenolic compounds, carotenoids and vitamins that act as free radicals scavengers, conferring antioxidant and antibacterial capacity to the matrix. These bioactive properties are related to the botanical origin of the honeybee pollen. Honeybee pollen samples were collected from different geographical locations in central Chile, and their total carotenoid content, polyphenols profile by HPLC/MS/MS, DPPH radical scavenging capacity, and antimicrobial capacity against S. pyogenes, E. coli, S. aureus, and P. auriginosa strains were evaluated. Our results showed a good carotenoids content and polyphenols composition, while antioxidant capacity presented values between 0-95 % for the scavenging effect related to the botanical origin of the samples. Inhibition diameter for the different strains presented less variability among the samples, Furthermore, binary mixtures representing the two most abundant species in each HBP were prepared to assess the synergy effect of the floral pollen (FP) present in the samples. Data shows an antagonist effect was observed when assessing the carotenoid content, and a synergy effect often presents for antimicrobial and antioxidant capacity for bee pollen samples. The bioactive capacities of the honeybee pollen and their synergy effect could apply to develop new functional ingredients for the food industry.

Honeybee Pollen From Southern Chile: Phenolic Profile, Antioxidant Capacity, Bioaccessibility, and Inhibition of DNA Damage.

Honeybee pollen (HBP) chemical composition is highly variable conforming to the floral and geographical origin of the pollen grains. The beneficial effects and functional properties of the HBP are well-known and have been mainly attributed to their high content of antioxidant polyphenols. In this work, twelve HBPs samples from the Southern region of Chile (X Región de Los Lagos) were characterized for the first time according to their botanical origin, phenolic composition, and antioxidant activity. The in vitro gastrointestinal digestion assay was done to simulate the human upper digestive tract. Selected honeybee pollen extracts (HBPEs) were assessed as bioaccessible fractions during an in vitro gastrointestinal digestion. Contents of phenolic compounds, antioxidant capacity, and recovery index of quercetin, myricetin, and cinnamic acid were monitored in different steps of gastrointestinal digestion. Furthermore, the protective effect of in vitro digested HBP towards DNA damage induced by peroxyl radicals was evaluated. The introduced species Brassica rapa L. (Brassicaceae), Lotus pedunculatus Cav. (Fabaceae), and Ulex europaeus L. (Fabaceae) predominated in all the HBPs analyzed, while the native species Buddleja globosa Hope (Scrophulariaceae), Luma apiculata (DC.) Burret (Myrtaceae), Embothrium coccineum J.R. Forst. & G. Forst. (Proteaceae) and Eucryphia cordifolia Cav. (Cunoniaceae) appeared less frequently. The content of polyphenols and antioxidant capacity in HBPEs achieved full bioaccessibility at the end of the intestinal digestion step. However, results obtained by a state-of-the-art technique (i.e. HPLC-DAD) demonstrated relatively low values of bioaccessible quercetin and cinnamic acid after the digestion process. In contrast, myricetin showed a high bioaccessibility in the intestinal digestion steps. The protective effect of in vitro digested HBP towards DNA damage induced by peroxyl radicals showed promising results (up to 91.2% protection). In conclusion, HBPs from the X Region de Los Lagos are rich sources of phenolic antioxidants that protect DNA from strand breakage. Therefore, the potential of HBPEs in preventing gastric and/or intestinal cancer should be further considered.

Biological activity of native Myrtaceae fruits from Chile as a potential functional food.

Myrtaceae fruits (Myrceugenia obtusa, Luma apiculata, and Luma chequen) were used as food and medicine by Chilean indigenous people. This study aimed to evaluate the bioactive properties of these berry-type fruits. The antioxidant capacity determined by the FRAP assay varied between 10.4 and 646.9 mmol Fe+2/g, while the antibacterial activity against Staphylococcus aureus and Salmonella typhi was 0 - 33 mm and 0 - 7.33 mm, respectively. All the extracts were rich in polyphenols and showed low cytotoxicity. Overall, M. obtusa presented dissimilar results compared to those of L. apiculata and L. chequen, encouraging the use of these native fruits as food, nutraceutical, or pharmacological ingredients.

Eucryphia cordifolia extracts: Phytochemical screening, antibacterial and antioxidant activities.

Eucryphia cordifolia is a native and dominant species from the Temperate Forest in the southern region of Chile. It is a crucial melliferous species producing unifloral honey with outstanding antibacterial properties with great commercial value at international markets. A phytochemical screening was developed by colorimetric and gravimetric methods. Antioxidant activities were assessed by using 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), and ferric ion reducing antioxidant power (FRAP) method. The minimum bactericidal concentration (MBC) was the method used to determine the antibacterial activity. The results revealed the highest antioxidant activity for the methanolic extracts of branches (IC50 35.7 µg/mL for DPPH, IC50 11.0 µg/mL for ABTS, and 4.6 mM FeSO4·7H2O/g in FRAP), attributed to its high content of total phenols (332.8 mg GAE/g). The quantification of the MBC led to values in the range of 0.78-12.5 mg/mL for S. aureus and S. pyogenes, and 6.25-50 mg/mL for E. coli and P. aeruginosa, respectively. The presented work provides valuable first in vitro evidence regarding the potential application of E. cordifolia extracts as antioxidants and antibacterial agents in the nutraceutical and phytopharmaceutical industry.

Bioactive compounds in Apis mellifera monofloral honeys.

Honey is a natural product with a sweet flavor. Honey is made by the honeybee (Apis mellifera L.) from the nectar of flowers or other plant secretions that are collected near the hive. These products are mixed with bee saliva and stored. Several studies have demonstrated that honey exhibits antioxidant, antimicrobial, nematicidal, antifungal, anticancer, and anti-inflammatory activities. These properties are influenced by the plants from which the secretions are harvested, from the naturally occurring compounds present in the nectar. Studies of the properties and applications of honey have distinguished honey from other natural products due to the presence of certain compounds and due its bioactive properties. The focus of this review is to discuss the identified and isolated compounds from monofloral honey produced by A. mellifera, with specific emphasis on antioxidant and antimicrobial properties of honey and its therapeutic health benefits.

Mayten Tree Seed Oil: Nutritional Value Evaluation According to Antioxidant Capacity and Bioactive Properties.

The Mayten tree (Maytenus boaria Mol.), a native plant of Chile that grows under environmentally limiting conditions, was historically harvested to extract an edible oil, and may represent an opportunity to expand current vegetable oil production. Seeds were collected from Mayten trees in north-central Chile, and seed oil was extracted by solvent extraction. The seed oil showed a reddish coloration, with quality parameters similar to those of other vegetable oils. The fatty acid composition revealed high levels of monounsaturated and polyunsaturated fatty acids. Oleic and linoleic acids, which are relevant to the human diet, were well represented in the extracted Mayten tree seed oil. The oil displayed an antioxidant capacity due to the high contents of antioxidant compounds (polyphenols and carotenoids) and may have potential health benefits for diseases associated with oxidative stress.

Ultrasound-Assisted Extraction of Anthocyanins Using Natural Deep Eutectic Solvents and Their Incorporation in Edible Films.

Extracts rich in bioactive compounds added to edible films have allowed the development of active packaging that increases the shelf life of food. However, it is necessary to search for solvents that are nontoxic and not harmful to the environment, with natural deep eutectic solvents (NADES) being an attractive and easily synthesized alternative. This research aimed to design NADES by lyophilization to be used in the extraction of anthocyanins from the Chilean Luma chequen (Molina) A. Gray berry, and subsequently adding them to the matrix of edible ƙ-carrageenan films. For this purpose, ultrasound-assisted extraction (UAE) was used and the anthocyanin content was evaluated with the pH differential method. The antioxidant capacity of extracts was determined by DPPH assay and the antibacterial capacity by diffusion agar tests. The results obtained indicate that the designed NADES are efficient at extracting anthocyanins, reaching concentrations between 81.1 and 327.6 mg eq cyanidin 3-glucoside/100 g dw of L. chequen (Molina) A. Gray. The extracts reached inhibition diameters between 5 and 34 mm against Escherichia coli, Staphylococcus aureus, and Salmonella typhi strains. Once the extracts were incorporated into ƙ-carrageenan films, active edible films with antioxidant and antibacterial capacities were obtained.

Antibacterial potential assessment of Nalca (Gunnera tinctoria Mol.) ethanolic extracts.

Nalca is an edible rhubarb-type plant from the southern of Chile; with studies focussing on petiole chemical and biological properties. This work evaluated for the first time the antibacterial capacities of three organs of nalca against Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. Petiole (P), root (R) and flower (F) ethanolic extracts at three different concentrations (25, 50, 100 mg/mL) were evaluated by means of agar diffusion assay and MIC. The result showed that antibacterial activities are bacteria and concentration dependent, with root having the highest antimicrobial activity of the organs plant studied. Compared to commercial antibiotics, nalca extracts show promising control over Ps. aeruginosa. The main polyphenolics of the extracts were identified by UPLC-MS/MS, observing for the first time caffeic, coumaric and gallic acids presence in all samples, with quercetin and rutin determined in petiole and flower extracts only. Our results contribute to better understanding of nalca attributes as potential source of antibacterial compounds to be used in nutraceutical, cosmetic and food industry.

Honeybee Pollen Extracts Reduce Oxidative Stress and Steatosis in Hepatic Cells.

Nonalcoholic fatty liver disease (NAFLD) is a major cause of morbidity and mortality worldwide. Additional therapies using functional foods and dietary supplements have been investigated and used in clinical practice, showing them to be beneficial. Honeybee pollen from Chile has shown a large concentration of phenolic compounds and high antioxidant activity. In this work, we characterized twenty-eight bee pollen extracts from the central zone of Chile according to botanical origin, phenolic profile, quercetin concentration, and antioxidant activity (FRAP and ORAC-FL). Our results show a statistically significant positive correlation between total phenolic content and antioxidant capacity. Selected samples were evaluated on the ability to reverse the steatosis in an in vitro cell model using Hepa1-6 cells. The pollen extracts protected Hepa1-6 cells against oxidative damage triggered by 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH)derived free radicals. This effect can be credited to the ability of the phenolic compounds present in the extract to protect the liver cells from chemical-induced injury, which might be correlated to their free radical scavenging potential. Additionally, bee pollen extracts reduce lipid accumulation in a cellular model of steatosis. In summary, our results support the antioxidant, hepatoprotective, and anti-steatosis effect of bee pollen in an in vitro model.

Antioxidant Effect of Extracts from Native Chilean Plants on the Lipoperoxidation and Protein Oxidation of Bovine Muscle.

The present study investigated the antioxidant potential and the ability to inhibit lipid and protein oxidation in bovine meat of four native Chilean species: canelo (Drimys winteri), nalca (Gunnera tinctoria), tiaca (Caldcluvia paniculata), and ulmo (Eucryphia cordifolia). Phenolic acids (gallic, chlorogenic, caffeic, and coumaric) and flavonoids (catechin, epicatechin, and rutin) were identified and quantified by HPLC-MS/MS. Drimys winteri extract exhibited the highest antioxidant capacity evaluated by oxygen radical absorption capacity-red pyrogallol method (ORAC-PGR) and ferric ion reducing antioxidant power (FRAP) assays. All extracts decreased lipid oxidation induced by 2,2'-azo-bis(2-amidinopropane) dihydrochloride (AAPH) derived peroxyl radicals by anywhere between 30% and 50% the. In addition, canelo and nalca extracts decreased spontaneous oxidation by around 57% and 37% in relation to the control group, being even more efficient than butylated hydroxyanisole (BHT) a synthetic antioxidant. Protein oxidation in the myofibrillar proteins was evaluated by the formation of protein carbonyls and loss of protein thiols. The canelo, ulmo, and nalca extracts decreased the formation of carbonyls by around 30%. Plant extracts and BHT did not show an antioxidant effect on protein thiol loss. However, tiaca and ulmo extracts exerted a pro-oxidant effect, favoring the oxidation of sulfhydryl groups. The oxidizing system induced structural changes in myofibrillar protein (SDS-PAGE). A protective effect on protein structure from the canelo extract can be observed during the incubation when compared to samples incubated with AAPH.

Honeybee Pollen Load: Phenolic Composition and Antimicrobial Activity and Antioxidant Capacity.

Honeybee pollen loads result from the agglutination of pollen grains and salivary secretions of bees. The potential use of honeybee pollen as a food supplement greatly depends on its chemical composition, which varies depending on the botanical and geographical origin of the pollen grains. This study aimed to characterize the botanical origin, chemical composition, and antioxidant and antibacterial activities of honeybee pollen from the V Region of Chile. The introduced species Brassica rapa and Eschscholzia californica predominated in the bee pollen analyzed. The honeybee pollen extracts showed antioxidant and antibacterial properties, specifically against the pathogenic microorganism Streptococcus pyogenes. Quercetin and myricetin were found in all samples in large concentrations. The separation of pollen loads from a multifloral sample demonstrated that E. californica pollen loads are responsible for antibacterial activity. This sample also showed a high concentration of quercetin (304.8 mg/100 g of bee pollen). Based on the present results, honeybee pollen from the V Region of Chile has been found to exhibit antioxidant and antimicrobial activities. Furthermore, it is proposed to use quercetin as a quality indicator for honeybee pollen from this region of Chile. These results should help establish better quality control criteria for Chilean honeybee pollen and its potential use as a functional ingredient.

LC-MS/MS analysis of neonicotinoid insecticides: Residue findings in chilean honeys / LC-MS/MS análises de inseticidas neonicotinoides: Resíduos encontrados em méis chilenos

ABSTRACT Neonicotinoids are a relatively new generation of insecticides that have been used for control of pests such as aphids, leafhoppers and whiteflies. This paper presents for the first time a determination of residues of four neonicotinoid insecticides (acetamiprid, thiamethoxam, thiacloprid and imidacloprid) in Chilean honey using QuEChERS extraction and UHPLC-MS/MS analysis. The limits of detection and quantification found for all analytes ranging from 0.34 to 1.43 μg kg-1 and from 0.30 to 4.76 μg kg-1, respectively. The extraction using QuEChERS method provided recoveries over 79% and the precision showed coefficient of variation lower than 20%. These data are in agreement with the international criteria that recommend general recovery limits of 70 - 120%. Of the 16 samples analyzed, in three honey samples neonicotinoids pesticides were detected. These three samples were collected from the same geographical area (Rengo). Fruit and grain production characterize the province of Rengo. The analysis of the botanical origin of these honeys showed the absence of pollen grains of crops and the majority presence of pollen grains of weeds such as Medicago sativa, Galega officinalis and Brassica rapa, which could be associated with crops. Although the residue levels found were low, the results also confirm the actual occurrence of a transfer of neonicotinoid insecticides from exposed honeybees into honey.

RESUMO Os neonicotinóides são uma geração relativamente nova de inseticidas que tem sido utilizado para o controle de pragas, como pulgões, cigarrinhas e moscas brancas. Este artigo apresenta pela primeira vez a determinação de resíduos de quatro inseticidas neonicotinóides (acetamiprida, tiametoxam, tiaclopride e imidaclopride) em méis chilenos empregando para extração o método QuEChERS seguido de UHPLC-MS/MS. Foram encontrados limites de detecção e quantificação aceitáveis para todos os analitos variando de 0,34 a 1,43 μg kg-1 e de 0,30 a 4,76 μg kg-1, respectivamente. A extração usando o método QuEChERS proporcionou uma recuperação maior que 79% e a precisão do método não excedeu um coeficiente de variação de 20%. Esses dados estão de acordo com os critérios internacionais que recomendam limites gerais de recuperação de 70 - 120%. Das 16 amostras de mel analisadas foram detectados pesticidas neonicotinoides em três delas. Essas três amostras foram coletadas na mesma área geográfica (Rengo), a qual se caracteriza pela produção de grãos e frutas. A análise da origem botânica desses méis mostrou a ausência de grãos de pólen de espécies cultivadas nesta zona, porém mostrou uma presença majoritária de grãos de pólen de ervas daninhas, como Medicago sativa, Galega officinalis e Brassica rapa, as quais podem estar associadas a estes cultivos. Embora os níveis de resíduos neonicotinoides encontrados neste estudo são baixos, os resultados confirmam a ocorrência atual de uma transferência destes inseticidas ao mel das de abelhas expostas.

Comparison of volatile compounds in Retanilla trinervia [Gillies & Hook] Hook & Arn honeys from central Chile / Comparación de compuestos volátiles en mieles de Retanilla trinervia[Gillies & Hook] Hook & Arn de Chile Central

The search for chemical markers for determining honey authenticity as a complementary tool for melissopalynological method is an important issue in the study of honeys from different botanical origins. The objective of this study was to determine the volatile compounds in tevo (Retanilla trinervia [Gillies & Hook] Hook & Arn [Fam. Rhamnaceae]) honey as one of the most relevant honeys from central Chile. For the identification and quantification of volatile compounds, Solid-Phase Microextraction and Gas Chromatography with Mass Spectrometry (SPME-GC-MS) was performed. A total of 28 out of 103 volatile compounds were found common to the five tevo honeys analyzed. Nevertheless, these compounds are common in unifloral honey of different floral sources from other countries. These results represent the first record in the identification of volatile compounds in tevo honey and would indicate that tevo honey does not present specific volatile compounds that allow its clear differentiation from other unifloral honey.

La búsqueda de marcadores químicos para determinar la autenticidad de la miel como una herramienta complementaria al análisis melisopalinológico es un tema importante en el estudio de las mieles de diferentes orígenes botánicos. El objetivo de este estudio fue determinar los compuestos volátiles en miel de tevo (Retanilla trinervia [Gillies & Hook] Hook & Arn [Fam. Rhamnaceae]), una de las mieles más relevantes de Chile Central. La identificación y cuantificación de compuestos volátiles se llevó a cabo mediante Microextracción en Fase Sólida y Cromatografía de Gases con Espectrometría de Masas (SPME-CG-MS). Un total de 28 de los 103 compuestos volátiles identificados en las cinco mieles de tevo analizadas se encontraron en común para ellas. Sin embargo, estos compuestos son comunes en la miel monofloral de diferentes fuentes florales de otros países. Estos resultados representan los primeros avances en la identificación de compuestos volátiles en la miel de tevo e indicarían que la miel de tevo no presenta compuestos volátiles específicos que permitan su clara diferenciación respecto a otras mieles monoflorales.

International Regulations of Propolis Quality: Required Assays do not Necessarily Reflect their Polyphenolic-Related In Vitro Activities.

Propolis has been proposed as a polyphenolic-rich natural product potentially able to be used for human consumption or even for medicinal proposes. To guarantee a minimum phenolic and flavonoid content and as consequence of their related-biological activities, international requirements of propolis quality are commonly applied. In this work we assessed phenolic and flavonoid contents of propolis; the antioxidant capacity (toward peroxyl radicals and hypochlorous acid); the ability to generate nitric oxide (NO); and, finally the antimicrobial activity of 6 propolis samples from the VI region of Chile. Our results show that the total phenolic and flavonoid content of propolis samples are not always in agreement with their polyphenolic-associated in vitro activities. For example, P03 and P06 samples showed the lowest (25 ± 4 GAE/g propolis) and the highest (105 ± 3 GAE/g propolis) total phenolic content, respectively. This was in agreement with flavonoid content and their Oxygen Radical Absorbance Capacity (ORAC) activity. However, this dependence was not observed toward HOCl, NO release and antimicrobial activity. Based on our results, we consider that, in order to guarantee the antioxidant or antimicrobial in vitro effects, the international regulations of propolis quality should contemplate the convenience of incorporating other simple analytical test such as ORAC or antimicrobial tests.

Chemical evidence for the liverwort complex, Chiloscyphus concavus and C. horizontalis.

During the phytochemical study of Chilean liverworts from Tierra del Fuego, two species were collected, Chiloscyphus concavus (Steph.) Hässel and C. horizontalis (Hook.) Nees. Their crude extracts, when analyzed by TLC and GC-MS, showed identical phytochemical profiles. In view of the macro- and micro-morphological differences used for the separation of both species, and the chemical evidence here presented, we conclude that variety is the more appropriate status for C. concavus.

Anthocyanin profiling of wild maqui berries (Aristotelia chilensis [Mol.] Stuntz) from different geographical regions in Chile.

BACKGROUND: Maqui (Aristotelia chilensis) is a Chilean species which produces small berries that are collected from the wild. Anthocyanins, because of their health benefits, are the major focus of interest in maqui fruit. For this study, we examined anthocyanin and phenolic content of maqui fruits from individuals that belonged to four geographical areas in Chile, and used DNA marker analysis to examine the genetic variability of maqui populations that had distinctly different fruit anthocyanin content. RESULTS: Twelve primers generated a total of 145 polymorphic inter simple sequence repeat-polymerase chain reaction (ISSR-PCR) bands. ISSR-PCR showed different banding patterns for the individuals evaluated, confirming that maqui populations belonged to different genotypes. Maqui fruit from four different geographical regions during two consecutive growing seasons showed high total anthocyanin (6.6-15.0 g cy-3-glu kg⁻¹ fresh weight (FW)) and phenolic (10.7-20.5 g GAE kg⁻¹ FW) contents and different anthocyanin profiles. CONCLUSION: Three maqui genotypes exhibited significantly higher anthocyanin content than the others, as measured by pH differential method and high-performance liquid chromatography-mass spectrometry. Significant genetic diversity was noted within each ecological population. ISSR-PCR analysis provided a fingerprinting approach applicable for differentiation of maqui genotypes.

Withanolides from Jaborosa caulescens var. bipinnatifida.

Withanolides 2,3-dihydrotrechonolide A (1) and 2,3-dihydro-21-hydroxytrechonolide A (2) were isolated along with two known withanolides trechonolide A (3) and jaborosalactone 39 (4) from Jaborosa caulescens var. bipinnatifida (Solanaceae). The structures of 1-2 were elucidated through 2D NMR and other spectroscopic techniques. In addition, the structure of withanolide 1 was confirmed by X-ray crystallographic analysis.