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The concentration of polyphenolic compounds and trace elements in the Coffea arabica leaves: Potential chemometric pattern recognition of coffee leaf rust resistance



by Eliana B. Souto 1,2,*,Alessandra Durazzo 3,Amirhossein Nazhand 4,Massimo Lucarini 3,Massimo Zaccardelli 5,Selma B. Souto 6,Amelia M. Silva 7,8,Patricia Severino 9,10,11,Ettore Novellino 12 andAntonello Santini 12,*

Francisco L.F.SilvaaGerlan O.NascimentobGisele S.LopesaWladiana O.MatosaRodrigo L.CunhacMarcelo R.MaltacGilberto RodriguesLiskadRobert W.OweneM. Teresa S.Trevisanba

Laboratório de Estudos em Química Aplicada (LEQA), Departamento de Química Analítica e Físico-Química, Centro de Ciências, Universidade Federal do Ceará, Campus do Pici, 60451-970 Fortaleza, CE, BrazilbDepartamento de Química Orgânica e Inorgânica, Centro de Ciências, Universidade Federal do Ceará, Campus do Pici, CP 12200, 60451-970 Fortaleza, CE, BrazilcEmpresa de Pesquisa Agropecuária de Minas Gerais (EPAMIG), Av. José Cândido da Silveira, 1657, 31170-495 Belo Horizonte, MG, BrazildUniversidade Federal do Pampa – UNIPAMPA, Rua Luiz Joaquim de Sá Britto, s/n, Bairro Promorar, 97650-000 Itaqui, RS, BrazileDivision of Preventive Oncology, National Center for Tumor Diseases, Im Neuenheimer Feld 460, and German Cancer Research Center (DKFZ), Im Neuenheimer Feld 581, 69210 Heidelberg, Germany

Received 16 October 2019, Revised 2 April 2020, Accepted 6 April 2020, Available online 21 April 2020, Version of Record 12 May 2020.


Coffee (Coffea arabica L.) is an important commodity, involving about 500 million people from the cultivation of the coffee trees to final consumption of infusions of the ground roasted coffee beans. In contrast to a considerable amount of research performed on green coffee beans, there are relatively few studies regarding the chemical constituents of coffee leaves. Hemileia vastatrix is a parasite, specific to coffee plants and causes coffee leaf rust, which is a very destructive disease. Some coffee plants have natural resistance which is mainly linked to a gene and specific host resistance response. An increase in flavonoid production may be related to fungal disease resistance, with the levels and flavonoid types being an early physiological response to rust infection. Trace inorganic elements can be related to many roles in the defense response of higher plants and can be used as a biomarker for some diseases. To address this, coffee leaves from 16 different cultivars of Coffea arabica were harvested from Minas Gerais, Brazil (susceptible and resistant to rust) and their polyphenolic compounds were extracted using the QuEChERS technique and quantitated by HPLC-ESI-MS. The same leaves were decomposed using an acid mixture in a block digester and the content of Al, Cu, Mg, Mn, Ni, Sn and Zn was quantitated by ICP-OES. Principal component analysis (PCA) was applied and we could establish a relation between polyphenolic and trace element concentration in the leaves with resistance to rust infection. On this basis in this preliminary study we were able to separate the resistant from the susceptible cultivars. The main compounds responsible for this differentiation were the content of chlorogenic acid and magnesium in the leaves. The content of polyphenolic compounds was lower in susceptible cultivars and a diametric effect was observed between Mn and Mg concentrations. This study shows potential for the discrimination of resistant and susceptible coffee trees based on the analyses of both trace element and polyphenolic concentration.

1. Introduction

Coffee is a product with mass consumption; an estimated 2.4 billion cups are consumed per day worldwide with an average of 2.4% annual growth over the last 10 years and it is one of the favorite beverages of the world (ICO, 2016).

Coffee (Coffea arabica L.) is an important commodity, involving about 500 million people from the cultivation of the coffee trees to final consumption of infusions of the ground roasted coffee beans (Batista et al., 2012). In contrast to a considerable amount of research performed on green coffee beans, there are relatively few studies concerning the chemical constituents of coffee leaves (de Almeida et al., 2014de Almeida et al., 2018Trevisan et al., 2016).

Hemileia vastatrix (H. vastatrix) is a parasite, specific to coffee plants and causes coffee leaf rust which is a very destructive disease. Some coffee plants have a natural resistance which is mainly linked to a gene and specific host resistance response (e.g·H2O2 production) (Fernandez et al., 2012). Coffea arabica is characterized by a low genetic diversity, which is therefore reflected in its susceptibility to numerous diseases.

Mechanical damage such as methyl jasmonate (MeJA) treatment, Hemileia vastatrix Berk. and Br. (H. vastatrix) fungus inoculation and L. coffeella infestation cause different responses in polyphenol oxidase (PPO) activity, and demonstrate that resistance may also be related to the oxidative potential of the tissue regarding the polyphenolic composition, rather than simply to a higher PPO activity (Fernandez et al., 2012Guerreiro-Filho, 2014Melo et al., 2006).

Mhlongo, Piater, Steenkamp, Madala, and Dubery (2014), discovered that phenolic compounds which prime plant defense, also induce the accumulation of chlorogenic acid, and other mono- and di-acylated caffeoylquinic acids.

Chlorogenic acids, present in leaves, act as antioxidants in plants and protect against degenerative, age-related diseases when present in the diet of humans. In plants, they can also modulate the feeding behavior of insects (Chen and Wu, 2014Mubarak et al., 2012).

Polyphenol oxidase (EC is a copper-containing enzyme responsible for hydroxylation of monophenols to o-diphenols and oxidation of o-diphenols to o-diquinones and is widely distributed in higher plants (Ramiro, Guerreiro-Filho, & Mazzafera, 2006). In this respect, 5-caffeoylquinic acid (chlorogenic acid) is regarded to be the best polyphenolic compound as a substrate of PPO present in coffee leaves (Mazzafera & Robinson, 2000). The levels of this compound are reported as higher in coffee leaf extracts of Coffea arabica than in coffee beans (de Almeida et al., 2018). At present, the most likely functions of PPO are its involvement in plant resistance to diseases (Constabel and Barbehenn, 2008Melo et al., 2006). PPO was investigated in compatible and incompatible interactions of coffee plants with the causal agent of coffee leaf rust (Melo et al., 2006).

A pathogenic defense role of PPO is also supported by studies showing good correlations between high PPO levels in coffee cultivars with high pathogen resistance (Niranjan Raj, Sarosh, & Shetty, 2006).

Mangiferin has also been shown to play a role in increasing the resistance of coffee trees to rust infection, a disease caused by H. vastatrix, which causes defoliation and drying of branches, notwithstanding a relation with the biochemical mechanisms of plant defense against pathogens and UV irradiation (Franklin, Conceição, Kombrink, & Dias, 2009).

An increase in flavonoid production may be related to resistance to fungal disease, with the levels and flavonoid types being an early physiological response to rust infection (de Oliveira Fassio et al., 2016).

There also may be a strong correlation between various factors which render Coffea arabica plants susceptible to rust infection. For example, (1) The concentration of trace elements (essential cofactors in the function of enzymes important in the deactivation of reactive oxygen species) such as PPO, superoxide dismutase and catalase (2) the concentration of major polyphenolic compounds.

To address this, coffee leaves harvested from Minas Gerais State, Brazil were extracted using the QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) technique, and polyphenol and trace element content (Al, Cu, Mg, Mn, Ni, Sn and Zn) were evaluated. Principal component analysis (PCA) was applied in an attempt to establish the role of polyphenolic and trace element concentration in the leaves to rust infection resistance. To our knowledge, this is the first study to attempt this approach.



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Alternative Health

Ivermectin – Niacin Research



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Alternative Health

A review on phytochemistry and medicinal properties of the genus Achillea



  • Received 30 Apr 2011; Revised 2 July 2011; Accepted 2 July 2011

1Saeidnia S., *1Gohari AR., 1Mokhber-Dezfuli N, 2 Kiuchi F.
1 Medicinal Plants Research Center, Faculty of Pharmacy, Tehran University of Medical
Sciences, Tehran, Iran. 2 Faculty of Pharmacy, Keio University, 1-5-30 Shibakoen, Minato-ku,
Tokyo 105-8512, Japan.


Achillea L. (Compositae or Asteraceae) is a widely distributed medicinal plant throughout the world and has been used since ancient time. Popular indications of the several species of this genus include treatment of wounds, bleedings, headache, inflammation, pains, spasmodic diseases, flatulence and dyspepsia. Phytochemical investigations of Achillea species have revealed that many components from this genus are highly bioactive. There are many reports on the mentioned folk and traditional effects. Although, the medicinal properties of Achillea plants are recognized worldwide, there are only one review article mainly about the structures of the phytochemical constituents of Achillea. The present paper reviews the medicinal properties of various species of Achillea, which have been examined on the basis of the scientific in vitro, in vivo or clinical evaluations. Various effects of these plants may be due to the presence of a broad range of secondary active metabolites such as flavonoids, phenolic acids, coumarins, terpenoids (monoterpenes, sesquiterpenes, diterpenes, triterpenes) and sterols which have been frequently reported from Achillea species.

Keywords: Achillea, Asteraceae, Bioactive compounds.

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The genus Achillea L. belongs to Asteraceae (Compositae), the largest family of vascular plants. Asteraceaeous plants are distributed throughout the world and most common in the arid and semi-arid regions of subtropical and lower temperate latitudes. Achillea contains around 130 flowering and perennial species and occurs in Europe and temperate areas of Asia and a few grow in North America. These plants typically have hairy and aromatic leaves and flat clusters of small flowers on the top of the stem. Since these flowers have various colors, a number of species are popular garden plants (14). The basic chromosome number of this genus is X=9 and most of the species are diploid with great ecological ranges from desert to water-logged habitats (5).

The name of Achillea is referred to the Achilles in the literary Trojan War of the Iliad who used yarrow to treat the soldiers’ wounds (6). The majority of the Achillea species are as the medicinal plants which have therapeutic applications (4). There are few review papers on the different aspects of Achillea as a noteworthy and medicinal genus. Recently, Si and co-authors (7) published a review article mainly about the structures of phytochemical constituents and a brief section of biological properties of Achillea (7). Literature reviews show that there are many reports on pharmacological, immunological, biological and other therapeutic activities of these valuable herbs which are reviewed in this article.

Traditional usages 

Since Achillea genus is widespread all over the world, its species have been used by local people as folk or traditional herbal medicines. Bumadaran is a popular name for several species of Achillea in Persian language. They are reported as tonic, anti-inflammatory, anti-spasmodic, diaphoretic, diuretic and emmenagogic agents and have been used for treatment of hemorrhage, pneumonia, rheumatic pain and wounds healing in Persian traditional literature (89).

In Spanish-speaking New Mexico and southern Colorado, A. millefolium L. is called plumajillo, or “little feather”, because of the shape of the leaves. Native Americans and early settlers used yarrow for its astringent qualities that made it effective in wound healing and anti-bleeding (10).

Achillea species are the most important indigenous economic plants of Anatolia. Herbal teas prepared from some Achillea species are traditionally used for abdominal pain and flatulence in Turkey (11). Dioscorides also used Achillea for dysentery, whether associated with cholera or other causes, which killed as many soldiers as did steel and lead. In terms of Chinese medicine, Achillea can be said to have three main actions: clear Exterior Wind (diaphoretic), Tonify Deficiency (tonic) and clear Heart Phlegm (anti-hypertention) (12).

Many of these therapeutic usages have been confirmed by new experimental and clinical studies. The consumption of herbal teas from different species of Achillea, especially for treatment of the gastrointestinal tract, is common in folk medicine (13). However, there are still several unknown aspects of Achillea plants that need more attention.

Phytochemical constituents 

Phytochemical investigations of Achillea species have revealed that many components from this genus are highly bioactive. The first anti-spasmodic flavonoids, cynaroside I and cosmosiin II (Scheme 1) were isolated from A. millefolium L. (14), and the first natural proazulene, achillicin III (Scheme 2) was identified from the genus Achillea (15). Literature search shows that the, flavonoids, terpenoids, lignans, amino acid derivatives, fatty acids and alkamides such as p-hydroxyphenethylamide IV (Scheme 2) have been identified in Achillea species. The main constituents of the most species have been previously reviewed (7). Therefore, in this article some other minor or rare compounds and especially their medicinal or industrial usages which have been less described are reviewed. Among them,alkamides, the lipophilic and nitrogen containing compounds, are responsible for insecticide, anti-inflammation and some immunological activities of Achillea and Echinacea plants (16). The genus Achillea comprises flavored species which produce intense essential oils. The volatile oils of Achillea contain monoterpenes as the most representative metabolites. However, there are reports on high levels of sesquiterpenes compared with monoterpenes (1718). There are several pharmacological actions which have been mostly attributed to the presence of azulenogenous sesquiterpene lactones in the essential oil of Achillea. Results of studies have indicated that tetraploid species are accumulating proazulenes such as achillicin III (Scheme 2) (19).Except for the essential oil constituents, yarrow (A. tenuifolia Lam.) seeds consist of the high oil content which is rich in linoleic acid, an essential polyunsaturated fatty acid. This makes yarrow seed as a potential source of edible oil for human consumption (20). Recently, A. millefolium has been introduced as a new source of natural dye for wool dyeing due to the presence of the flavonoids, luteolin V and apigenin VI (Scheme 1). A. millefolium was found to have good agronomic potential as a natural dye in Iran (21). In the plant kingdom, hydroxycinnamoyl conjugates of quinic acid represent common end metabolites of the shikimate-phenylpropanoid pathway, and feruloylcaffeoylquinic acid derivates VII have been isolated only from two species of genus Achillea so far (22). From the aerial parts of Achillea species, proline VIII, stachydrine IX, betonicine X, betaine XI and choline XII have been isolated as the major nitrogen containing compounds (Scheme 2) (2324). Betaines, containing the permanent positive charge on the quaternary ammonium moiety, belong to an important class of naturally occurring compounds that function as compatible solutes or osmoprotectants (25). These compounds have shown immunosuppressive activity in the experimental animals (2627).



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Yerba Mate (Ilex paraguariensis) Beverage: Nutraceutical Ingredient or Conveyor for the Intake of Medicinal Plants? Evidence from Paraguayan Folk Medicine



Monika Kujawska


The use of medicinal plants mixed with yerba mate (Ilex paraguariensis) has been poorly studied in the ethnopharmacological literature so far. The Paraguayan Mestizo people have the longest tradition of using the yerba mate beverage, apart from the indigenous Guarani people. This study analyses the role of yerba mate and medicinal plants in the treatment of illnesses within Paraguayan folk medicine. The research was conducted among 100 Paraguayan migrants living in Misiones, Argentina, in 2014 and 2015. Yerba mate is not considered to be a medicinal plant by its own virtues but is culturally a very important type of medicinal plant intake. Ninety-seven species are employed in hot and cold versions of the yerba mate beverage. The most important species are as follows: Allophylus edulis (highest number of citations), Aristolochia triangularis (highest relative importance value), and Achyrocline flaccida and Achyrocline tomentosa (highest score by Index of Agreement on Species). The plants are used in the treatment of 18 medicinal categories, which include illnesses traditionally treated with plants: digestive system, humoral medicine, and relatively new health conditions such as diabetes, hypertension, and high levels of cholesterol. Newly incorporated medicinal plants, such as Moringa oleifera, are ingested predominantly or exclusively with the mate beverage.

1. Introduction

Yerba mate (Ilex paraguariensis A.St.-Hil., Aquifoliaceae) is a native tree growing in the subtropics of South America, present in Southern Brazil, Northeastern Argentina, Eastern Paraguay, and Uruguay [1]. The yerba mate beverage has been consumed traditionally by Guarani indigenous people since before the conquest of South America by the Spaniards [2]. The commercial potential of this plant was discovered by the Jesuits, who brought wild growing yerba mate into cultivation. Pedro de Montenegro, a Jesuit monk, in his Materia Medica Misionera described the use of the most important species for the Guarani people, in which yerba mate appeared on the top of the list [3]. The Guarani name for yerba mate is ka’a which means “a plant” or “a herb”; hence yerba mate has been considered by this group as the plant par excellence [3]. Yerba mate was also known as Jesuit tea or Paraguayan tea and shipped as such to Europe [2]. With the expulsion of the Jesuits in 1768, the plantations went wild. By this time, the yerba mate beverage was already popular among Mestizo people (of Spanish and Guarani origin). Since the end of the 19th century, it also became a daily beverage for the European migrants who partly colonized Southern Brazil, Northeastern Argentina, and, to a lesser extent, Eastern Paraguay [4]. Nowadays yerba mate is consumed at the rate of more than one litre per day by millions of people in the above-mentioned countries [45]. It plays a very special social role and constitutes a very important form of caffeine intake [245]. Its popularity is also increasing outside South America due to its pharmacological properties, proven to be beneficial to health [467]. It is also a very important drink in Syria and Lebanon due to Syro-Lebanese migration to Argentina in the second half of the 19th century. Many migrants who returned to the Levant in the 1920s took the habit of drinking mate with them [89].

Over the last 20 years there has been an increase in studies of the pharmacologic properties of Ilex paraguariensis, which have been reviewed [46710]. Numerous active compounds have been identified in yerba mate. Phenolic compounds predominate caffeoyl derivatives (caffeic acid, chlorogenic acid) [1112], xanthines (caffeine and theobromine), which are a class of purine alkaloids found in many other plants such as tea and coffee, flavonoids (quercetin, kaempferol, and rutin), and tannins [7]. Numerous triterpenoid saponins have also been identified, including those derived from ursolic acids known as metasaponins [47]. Saponins are responsible for the distinct flavour of yerba mate extracts [7]. Yerba mate also contains minerals (P, Fe, and Ca) and vitamins (C, B1, and B2) [13].

Research on extracts and isolated compounds from yerba mate has provided a number of pharmacological applications. Studies have demonstrated that yerba mate leaves have antioxidant [11], antiobesity [1415], antidiabetic, digestive improvement and cardiovascular properties [1617], and chemopreventative ones (preventing cellular damage that may cause chronic diseases) [18]. The consumption of yerba mate infusion reduces LDL-cholesterol in parallel with an increase in HDL-cholesterol, as observed in studies on humans [19]. Yerba mate extract also reduces acute lung inflammation, as observed in the animal model [4]. Antimicrobial activity of Ilex paraguariensis has been recently studied as well [20].

Some ethnobotanical studies from the south cone of South America report medicinal uses of yerba mate beverage [2122]. Few ethnobotanical and ethnopharmacological studies mention that various medicinal plants are consumed together with the yerba mate beverage by Mestizo and European migrants living in Argentina and Paraguay [2326]. However, very little is known about how medicinal plants are combined with yerba mate beverage by local people. Additionally, medicinal plant use by Paraguayan Mestizo people is poorly documented in the English-language scientific literature, with very few exceptions [232630]. The documentation of medicinal plants and analysis of traditional knowledge related to the yerba mate beverage by Paraguayan Mestizo people is of paramount importance for two reasons: (1) apart from indigenous Guarani peoples, they have the longest tradition of using yerba mate and mixing it with medicinal plants; (2) The Paraguayan people are described in the literature as knowledgeable about medicinal plants [3031]. Nearly 80% of the population of Paraguay consume medicinal plants on a daily basis [30]. However, the relationship between traditional uses and pharmaceutical properties is poorly studied.

The objectives of this contribution were to (1) document and analyse the role of yerba mate in prophylaxis and treatment by Paraguayan Mestizo people; (2) evaluate the role of medicinal plants in yerba mate beverages, and (3) describe the scope of illnesses treated with yerba mate beverage and medicinal plants. Additionally, two questions guided my research and analysis: (1) Does any pattern exist showing that particular illnesses are treated with a hot version of yerba mate beverage and others with a cold one? (2) How receptive is this traditional mode of plant administration to new health challenges and new medicinal plants, previously unknown to the Paraguayan people?



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