Investigation of biologically active amines in some selected edible mushrooms

Journal Pre-proof Investigation of biologically active amines in some selected edible mushrooms ´ ´ Guilherme C.L. Reis, Flavia B. Custodio, Bruno G. Botelho, Let´ıcia R. Guidi, Maria Beatriz A. Gloria

PII:

S0889-1575(19)31001-4

DOI:

https://doi.org/10.1016/j.jfca.2019.103375

Reference:

YJFCA 103375

To appear in:

Journal of Food Composition and Analysis

Received Date:

6 July 2019

Revised Date:

3 November 2019

Accepted Date:

11 November 2019

´ Please cite this article as: Reis GCL, Custodio FB, Botelho BG, Guidi LR, Gloria MBA, Investigation of biologically active amines in some selected edible mushrooms, Journal of Food Composition and Analysis (2019), doi: https://doi.org/10.1016/j.jfca.2019.103375

This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2019 Published by Elsevier.

Investigation of biologically active amines in some selected edible mushrooms

Guilherme C.L. Reisa, Flávia B. Custódioa, Bruno G. Botelhob, Letícia R. Guidic, Maria Beatriz A. Gloriaa*

a

LCQ – Laboratório de Controle de Qualidade, Faculdade de Farmácia, Universidade

Federal de Minas Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brasil Departamento de Química, Instituto de Ciências Exatas, Universidade Federal de Minas

ro of

b

Gerais, Av. Antônio Carlos 6627, Belo Horizonte, MG, 31270-901, Brasil c

Faculdade de Engenharia Química, Universidade Federal de Uberlândia, Campus

-p

Avançado Patos de Minas, Rua Padre Pavoni 290, Patos de Minas, MG, CEP 38701-002,

re

Brasil

lP

* Corresponding author: E-mail address: [email protected] (M. Beatriz A. Gloria). Telephone: +55 31 3409 6926

Chemical compounds studied in this article

na

Spermidine (PubChem CID: 9539); Agmatine (PubChem CID: 2794990); Putrescine (PubChem CID: 9532); Cadaverine (PubChem CID: 80282); Histamine (PubChem CID:

ur

5818); Tyramine (PubChem CID: 66449); Phenylethylamine (PubChem CID: 9075); Tryptamine (PubChem CID: 67652); Serotonin (PubChem CID: 160436)

Jo

Graphical abstract

1

ro of -p re

Highlights

A HPLC method was validated for the analysis of nine bioactive amines in

lP



mushroom.

Spermidine was the predominant amine quantified in all mushrooms.



Agmatine, a precursor of polyamines, was inherent to Pleurotus spp.



Multivariate analysis of amines differentiated Pleurotus spp. and Agaricus spp.

Jo

ur

na



ABSTRACT (194/200) Mushrooms are highly valued due to nutritional and functional properties as well as small environmental footprint. However, scarce information is available regarding amines in commercial products. The objective of this study was to investigate the levels of bioactive 2

amines in eight fresh edible commercial mushrooms species.

An ion-pair HPLC method

with post-column derivatization with o-phthalaldehyde and fluorescence detection was fit for the purpose. Seven out of nine amines were present and levels varied among species. Spermidine was ubiquitous to mushrooms, with highest content in Black Shimeji (12.4 mg/100 g). The levels of spermidine in mushrooms classify them as high polyamines sources, which is valued due to its association with growth, health promotion and antioxidant properties. Agmatine was present in all Pleurotus. Tyramine, tryptamine and phenylethylamine were detected in some species; the levels of cadaverine and putrescine

ro of

were discrete. A four-principal component model explained 99.4% of the variance and it was able to separate Pleurotus spp. (White shimeji, Hiratake, Black shimeji and Salmon) from Agaricus bisporus (Champignon and Portobello) and Lentinula edodes (Shitake).

Keywords:

lP

re

amines to separate some mushroom species.

-p

Hierarchical cluster analysis confirmed the potential of using the occurrence and levels of

Principal component analysis; Hierarchical cluster analysis; Spermidine; Agmatine;

na

Polyamines; Biogenic amines.

Jo

ur

.

3

1. Introduction

Edible mushrooms are widely consumed in many countries due to nutritional value, low calories and specific aroma and texture, a delicacy (Kalač, 2013; Feeney et al., 2014). The consumption of mushrooms has increased throughout the world. According to the Food and Agriculture Organization (FAO) of the United Nations, cultivated mushroom worldwide production increased from 5.91 in 2007 to 10.24 million tons in 2017, which represents an increment higher than 73% in ten years (FAOSTAT, 2019).

Besides,

ro of

several types of mushrooms are available, both cultivated/commercial and wild noncommercial edible species. In addition to the economic value, mushrooms have a small environmental footprint, as they grow from agricultural and forest wastes and require

-p

relatively little water or land. Furthermore, they can be used as agents of environmental

re

management, by using natural resources in a less destructive way for the biosphere and by promoting sustainable development in all ecosystems (Donnini et al., 2013; Feeney et

lP

al., 2014). According to the Associação Nacional dos Produtores de Cogumelos – ANPC (Mushrooms Producers National Association), it is estimated that Brazil produces over 12

na

thousand tons of fresh mushrooms every year (ANPC, 2019). The most popular edible mushroom species available are Agaricus bisporus, Lentinula edodes and some Pleurotus

ur

species.

Due to their chemical composition, edible mushrooms constitute a food of nutritional They have low calories (low fat contents) and high contents of minerals (K),

Jo

value.

essential amino acids, vitamins (provitamin D2, vitamin B12) and fiber (Roupas et al., 2012; Kalač, 2013; Feeney et al., 2014; Manninen et al., 2018). They are important sources of polysaccharides with immunomodulating properties.

In addition, mushrooms contain

several natural phytochemicals with a wide range of positive health effects and medicinal benefits (Roupas et al., 2012).

Some preclinical and clinical studies suggest positive 4

impacts of mushrooms on brain health and cognition, weight management, oral health, constipation, diabetes, anti-arthritic and anti-viral agents (Roupas et al., 2012; Feeney et al., 2014; Pop et al., 2018). Preliminary evidence suggests that mushrooms may support healthy immune and inflammatory responses through interaction with gut microbiota, enhancing the development of adaptive immunity and improved immune cell functionality (Roupas et al., 2012; Feeney et al., 2014). Many of the immunomodulation effects attributed to mushrooms are due to polysaccharide, either β-glucans or polysaccharide-protein complexes (Roupas et al.,

ro of

2012). However, part of the therapeutic properties is related to their inhibition of oxidative stress (Yang et al., 2002; Elmastas et al., 2007; Silva & Jorge, 2014; Janjušević et al., 2017; Pop et al., 2018). The antioxidant activity of mushrooms may be attributed to the

-p

presence of several components, such as phenolics (Janjušević et al., 2017; Pop et al.,

re

2018) and bioactive amines (Rider et al., 2007; Toro-Funes et al., 2013). Reports on the occurrence and levels of bioactive amines in mushroom are scarce,

lP

especially commercial ones. Turkish wild-growing mushrooms were reported to be high sources of spermidine, followed by putrescine, tyramine, tryptamine and phenylethylamine

na

(Dadáková et al., 2009). Commercial mushrooms have been analyzed mainly for the presence of polyamines (Okamoto et al., 1997; Cipolla et al., 2007; Nishibori et al., 2007).

ur

Only a few studies investigated additional amines in commercial mushroom including putrescine (Okamoto et al., 1997), putrescine and cadaverine (Yamamoto et al., 1982) and

Jo

tryptamine, 2-phenylethylamine, histamine and tyramine (Yen, 1992). In this context the profile and levels of bioactive amines in eight fresh edible

mushrooms species commercialized in Brazil were analyzed for the first time. A HPLC method for the determination of nine amines in mushroom was validated. The potential functional properties of the mushrooms due to bioactive amines were described. Furthermore, the possibility of using amines as criterion for clustering different mushrooms 5

species was investigated by multivariate analysis (principal component analysis - PCA and hierarchical and clustering analysis - HCA).

2. Material and Methods

2.1. Reagents and solvents

The reagents used were of analytical grade, except HPLC solvents (acetonitrile and For HPLC analysis, the organic and

ro of

methanol) which were chromatographic grade.

aqueous solvents were filtered through 0.45 µm pore size HAWP and HVWP membranes,

Q Plus System (Millipore Corp., Milford, MA, USA).

-p

respectively (Millipore Corp., Milford, MA, USA). Deionized water was obtained from Milli-

re

Spermine (SPM) tetrahydrochloride, spermidine (SPD) trihydrochloride, putrescine (PUT) dihydrochloride, cadaverine (CAD) dihydrochloride, tyramine (TYM) hydrochloride, (HIM)

dihydrochloride,

agmatine

(AGM)

sulphate,

serotonin

(SRT)

lP

histamine

hydrochloride, 2-phenylethylamine (PHM) hydrochloride, tryptamine (TRM) free base, and

ur

2.2. Samples

na

o-phthalaldehyde (OPA) were purchased from Sigma Chemical Co. (St. Louis, MO, USA).

Jo

Fresh mushrooms were purchased at commercial maturity from producers within the

region of Belo Horizonte, state of Minas Gerais, Brazil, among them, Pleurotus spp. – Black shimeji, White shimeji, Hiratake, Salmon and Eryngii; Agaricus bisporus – Champignon and Portobello; and Lentinula edodes – Shiitake (Table 1). Different lots of each mushroom species (≥ 200 g) were obtained, except for Eryngii which, because of its limited availability, had only two lots collected. All the lots were obtained from the same 6

producer, which is the major provider for commercial use, immediately after harvest. The fresh mushrooms were homogenized, grated and analyzed immediately for moisture (AOAC, 2012) and bioactive amines contents.

2.3. Method for the determination of bioactive amines

Nine free bioactive amines (spermidine, agmatine, putrescine, cadaverine, tyramine,

ro of

tryptamine, phenylethylamine, histamine and serotonin) were determined according to the method described by Bandeira et al. (2012). Briefly, the amines were extracted from 3 g fresh mushrooms with 5% trichloroacetic acid. The samples were agitated for 5 min on a

-p

shaker (250 rpm) and centrifuged at 8422 g for 20 min at 4 °C.

These steps were

re

repeated two more times. The supernatants were combined, filtered through qualitative filter paper and through HAWP membrane (0.45 µm pore size, Millipore Corp., Milford, MA,

lP

USA) and used for analysis.

The amines were separated by ion-pair HPLC and quantified, after post column

na

derivatization with o-phthalaldehyde, fluorometrically at 340 nm excitation and 450 nm emission. A Shimadzu LC-10AD with SIL-20AHT automatic injector (Shimadzu, Kyoto,

ur

Japan) connected to a RF-510 AXL spectrofluorometric detector and to a CBM-20A controller was used. The column and pre-column used were Novapack® C18 (3.6 x 300

Jo

mm, 4 µm) (Waters, Milford, MA, USA).

Two mobile phases were used: (A) sodium

acetate buffer (pH 4.9) with 15 mmol/L sodium octanosulphonate and (B) acetonitrile, at a flow rate of 0.8 mL/min, in a gradient elution: initial to 21.0 min/3–20% B; 21.0 to 22.0 min/20–5% B; 22.0 to 25.0 min/5% B; 25.0 to 40.0 min/5-24% B; 40.0 to 45.00 min/24% B; 45.0 to 50.0 min/24–35% B, 50.0 to 51.0 min/ 35–3% B and further re-equilibration at initial

7

conditions for another 9.0 min, in a total cycle time of 60.0 min until the next injection. The injection volume was 10 µL.

2.4. Intralaboratory validation of the method

The fitness of the method described by Bandeira et al. (2012) was evaluated for the mushroom matrix according to the following parameters: specificity, linearity of calibration curves, accuracy, precision and limits of detection and quantification (Thompson et al., Agaricus bisporus mushrooms were chosen for the validation process.

ro of

2002).

The

specificity of the method was determined by injection of 20 different mushroom extracts. The existence of any interference (possible peaks) that could affect detection in the range

The samples were also spiked with each single standard to

confirm the identity of each peak.

re

nine amines standards.

-p

of retention time of the target analytes was investigated. The samples were spiked with

The calibration curves were constructed by using

lP

different concentrations of all amines (0.1, 0.2, 1.0, 2.0, 4.0, 6.0, 8.0, 10.0, 12.0 μg/mL) injected randomly. Repeatability and reproducibility were evaluated by analysis of spiked

na

samples (4 mg/100 g) in five replicates and by three different analysts. Accuracy was calculated as the mean concentration found divided by the fortification level and multiplied

ur

by 100. Repeatability was established through determination of the coefficient of variation. The analyses were carried out at three different days with three different analysts to

Jo

evaluate reproducibility.

Mean concentration, standard deviation and coefficient of

variation were calculated for the spiked samples of each analyst. The limit of detection was the lowest concentration of the analyte corresponding to three times the signal-tonoise ratio. The limit of quantification was the lowest concentration of the analyte that could be determined with acceptable accuracy and precision. It was considered the first analyte concentration at the calibration curve (Thompson et al., 2002). 8

2.5. Statistical analysis

The results were submitted to Shapiro Wilk test for normality and Levene’s test for homoscedasticity. Then, the data was submitted to analysis of variance and the means were compared by the Tukey test at 5% probability (Granato et al., 2014) using the Past 3.19 software (UIO, Oslo, Norway).

ro of

2.6. Multivariate analysis

Two multivariate exploratory techniques – Principal Component Analysis (PCA) and

-p

Hierarchical Cluster Analysis (HCA) – were applied for the characterization of mushrooms

re

in relation to the profile and level of bioactive amines (Granato et al., 2018a). PCA was used to project multivariate data onto a smaller dimensional space, without affecting the Consequently, relevant information was separated and

lP

relation between samples.

amplified, making them more perceivable by visual inspection, and allowing the discovery,

na

visualization and interpretation of differences between variables, the relation between samples, and the identification of outliers. HCA was used as a tool for dimensionality

ur

reduction, by clustering the objects by similarity, minimizing intragroup variance and maximizing intergroup variance.

HCA results were presented in bidimensional plots

Jo

(dendrograms), representing the hierarchical structure of the data, in which the length of each branch represents similarity between samples (Alves et al., 2010; Otto, 2017; Granato et al., 2018b). Both PCA and HCA models were built using PLS ToolBox 6.5 (The Eigenvector Technologies, Manson, WA, USA) and MATLAB software, version 2010a (The MathWorks, Natick, MA, USA).

9

3. Results and discussion

3.1. Method validation

The fitness of the method for the quantification of bioactive amine in mushrooms is presented in Table 2.

Regarding linearity, the calibration curves had coefficients of

determination (R2) equal or higher than 0.9966 indicating a high correlation between the concentration and the signal area. By visual inspection, good distribution of the residues

ro of

was observed, which was confirmed by the tests of independence (Durbin-Watson) (pvalue > 0.05) and homoscedasticity (Breuch-Pagan) (p-value > 0.05). Thus, the linear equations for the calibration curves are valid for the quantification of each of the nine

-p

amines in mushrooms in the range of 0.096 to 12.0 mg/100 g.

The accuracy of the

re

method was evaluated by the recovery of the analytes in spiked samples, and ranged from 70.7% (serotonin) to 104.4% (spermidine). The intraday precision (repeatability) of the

lP

method ranged from 4.7% (histamine) to 6.5% (putrescine) and the interday precision (reproducibility) ranged from 6.7% (phenylethylamine) to 15.3% (serotonin). Therefore,

na

the method was accurate and precise for the analysis of nine amines (spermidine, agmatine, putrescine, cadaverine, tyramine, phenylethylamine and tryptamine) in

ur

mushrooms. The limits of detection ranged from 0.029 to 0.032 mg/100 g and the limits of quantification ranged from 0.096 to 0.105 mg/100 g (Table 2), which confirms the good

Jo

sensitivity of the method. These results indicate that the method is fit for the analysis of amines in mushroom.

The use of on line derivatization and fluorescence detection

improves selectivity and sensitivity of the method, which certainly contributed to the success of the method (Önal et al., 2013).

10

3.2. Occurrence of amines in the mushrooms

Among the nine amines investigated, seven were detected in different mushroom species but in a distinct way (Table 3). Histamine and serotonin were not detected in any of the mushroom species studied.

Spermidine was present in all mushroom species,

suggesting that it is inherent to mushrooms. Agmatine was detected in six out of the eight mushroom species; it was present in every Pleurotus spp. mushrooms investigated, suggesting that agmatine, along with spermidine, can be inherent to Pleurotus spp.

ro of

Tryptamine was also present in 100% of the samples of White shimeji, suggesting that it is ubiquitous to this mushroom specie.

-p

It is interesting to observe that Portobello had only one amine (spermidine), followed

re

by Eryngii which had two (spermidine and agmatine). However, White shimeji, Hiratake and Salmon had seven of the nine amines investigated.

lP

The occurrence of spermidine in mushroom, as well as in all living cells, has been reported in the literature for wild mushrooms (Okamoto et al., 1997; Dadáková et al., 2009;

na

Kalač, 2013) and its presence is associated with several relevant roles on cellular metabolism and growth.

Agmatine was previously detected at very low levels in

ur

honshimeji mushroom (Okamoto et al., 1997). Its presence in mushrooms suggests an additional route for the synthesis of polyamines via arginine (Bandeira et al., 2012).

Jo

Although putrescine is an obligatory precursor of polyamines, it was only detected in Pleurotus spp. Therefore, despite being relevant in the formation of polyamines, it does not seem to accumulate in some types of mushroom, or it is present at low concentrations, below the limit of detection of the method. Unlike the commercial mushrooms currently studied, putrescine was the predominant amine in wild-growing mushrooms (Dadáková et al., 2009).

Tyramine, tryptamine and phenylethylamine may be inherent to some 11

mushroom species, as secondary metabolites, or they may result from the decarboxylation of the precursor amino acids (tyrosine, tryptophan and phenylalanine, respectively). They were also detected in wild-growing mushrooms (Dadáková et al., 2009). The presence of cadaverine in mushroom was reported by Okamoto et al. (1997); however, they failed to separate cadaverine from histamine and quantified both of them (histamine/cadaverine) together. Histamine was not detected in the mushrooms analyzed in this work, similar to the results found by Dadáková et al. (2009) for wild-growing

ro of

mushrooms.

3.3. Levels of amines in the mushrooms

-p

The mean total levels of free bioactive amines in the mushrooms varied from 7.77 up

re

to 36.6 mg/100 g (Table 3), with the highest levels found in Salmon. Lower contents were observed in Shiitake (L. edodes), Portobello and Champignon (≤10 mg/100 g) – A.

lP

bisporus, whereas intermediate levels (from 10 to 25 mg/100 g) were found in Eryngii, Black shimeji, White shimeji and Hiratake (Pleurotus spp.).

na

Spermidine contributed the most (≥ 98.6%) in Portobello and Champignon (Agaricus bisporus) and also in Shiitake (Lentinula edodes, 92.8%); however, in Pleurotus spp., the

ur

contribution varied from 31.4 to 77%. Agmatine contributed with 23 to 57% to the total levels in Pleurotus spp. and with 6.3% to the total levels in Shitake. Indeed, in some

Jo

Pleurotus spp. mushrooms, the contribution of agmatine was higher compared to spermidine, e.g. White shimeji, Hiratake and Salmon. The other amines contributed with less than 5%, except for putrescine in Salmon (6.8%) and tryptamine in White shimeji (6.7%). The concentration of spermidine in the mushrooms ranged from 3.23 to 17.2 mg/100 g (Table 4).

There was significant difference on the levels of spermidine among 12

mushrooms, with the highest levels in Black Shimeji (12.4 mg/100 g) and the lowest in White shimeji (7.00 mg/100 g). Based on the classification proposed by Kalač (2014), the mushrooms analyzed in this study can be considered as high (> 1 mg/100 g) or very high (> 10 mg/100 g) sources of polyamines. Such polyamines levels can provide various health benefits, including promotion of intestinal health (Kalač, 2014; Ramani et al., 2014; Rogers et al., 2015), development of the immune system, wound healing, antiinflammatory and antioxidant agents and cardioprotective effects (Kalač, 2014; Ramani et al., 2014; De Cabo & Navas, 2016; Handa et al., 2018; Sharma et al., 2018). In spite of

ro of

this, polyamines should be avoided by individuals with cancer as they may increase tumor growth (Kalač, 2014; Ramani et al., 2014).

-p

Higher mean levels of agmatine (20.9 mg/100 g) were found in Salmon; followed by

decarboxylation of arginine.

re

the other Pleurotus spp. mushrooms; and, them, by Shitake. Agmatine is formed from the Recently, agmatine has been associated with beneficial

lP

health effects, among them, neuroprotection in the central nervous system, in mental illness, in depression and schizophrenia (Laube & Bernstain, 2017).

na

With respect to putrescine, even though it is an obligatory precursor of the polyamines, it was not detected in Agaricus bisporus and in Lentinula edodes. It was

ur

found only in three of the five Pleurotus spp. mushrooms. Higher mean levels were found in Salmon and White shimeji. These levels are lower compared to the values reported for

Jo

wild-growing mushrooms (Dadáková et al., 2009). In a similar way, cadaverine levels were low (≤ 2.72 mg/100 g) and it was only detected in the same mushrooms which contained putrescine. It is well known that both putrescine and cadaverine can contribute with putrefactive flavor to foods; however, the levels found are 78% lower than the threshold values reported for putrescine and cadaverine – 109 mg/kg (Wang et al., 1975).

13

Tyramine was detected in all mushroom species, except Portobello and Black shimeji. It was not present in all the lots of mushrooms analyzed, which suggests that its formation is affected by extrinsic factors during production, including stress and substrate composition.

No significant difference was found among mean levels for different

mushroom species.

Tryptamine was detected in most mushroom species, except for

Portobelo and Eryngii. Higher mean levels (p < 0.05) were found in White shimeji (1.37 mg/100 g), followed by Salmon (0.82 mg/100 g) and, then, by the other types (0.05 – 0.18 mg/100 g). All analyzed lots of White shimeji and Salmon contained tryptamine, which

ro of

could suggest that this amine is inherent to these mushrooms. Phenylethylamine was detected only in Pleurotus spp., except for Black shimeji and Eryngii. Levels were lower than 1.91 mg/100 g, with highest levels in White shimeji.

Tyramine, tryptamine and

-p

phenylethylamine were also detected at similarly low levels in wild-growing mushrooms

re

(Dadáková et al., 2009). They were detected in commercial mushrooms for the first time. Tyramine and phenylethylamine are neuroactive compounds. At low concentrations,

lP

they can exert important roles such as neuromodulation and mood-lifting effects. However, at high concentrations, tyramine and tryptamine can cause adverse effects to

na

human health and are, therefore, of food safety concern. Hypertension and symptoms such as headache, vomiting, perspiration, pupil dilatation and migraine have been

ur

described (EFSA, 2011). There is still limited information on the safe levels of tryptamine. As for tyramine, the levels in mushrooms were ≤ 5.98 mg/100 g, which are below the no

Jo

adverse health effect level established for healthy individuals (600 mg per meal); but the consumption of Hiratake could cause adverse effects to individuals taking classical monoamine oxidase inhibitor (MAOI) drugs (NOAEL = 6 mg per meal) (EFSA, 2011).

3.4. Multivariate analysis

14

Multivariate analysis of autoscaled data indicated that a four-principal component (PC) model explained 99.4% of the variance.

PC1 explained 59.4% of the variance

(Figure 1a) and it separated White Shimeji, Hiratake and Salmon mushrooms (positive PC1 values) from the other mushrooms. According to PC1 loadings (Figure 1c), all of the amines contributed to this differentiation (positive PC1 values), but with various intensities. Putrescine, cadaverine, agmatine and total amines content had a higher impact than tyramine, spermidine and phenylethylamine.

These three mushrooms (White Shimeji,

Hiratake and Salmon) belong to the same genus – Pleurotus spp. Eryngii and Black

ro of

shimeji also belong to this genus, but they differed from the others as most of the biogenic amines were not detected.

-p

PC2 explained 24.4% of the variance and it basically separated White Shimeji

re

(positive values) from all the other types of mushrooms (Figure 1a), mainly because of the concentrations of phenylethylamine and tryptamine – positive values, and spermidine –

lP

negative values (Figure 1c). In fact, according to Table 2, White Shimeji had the highest concentrations of phenylethylamine and tryptamine (1.37 and 0.84 mg/100 g, respectively)

na

and the lowest spermidine concentration (7.00 mg/100 g). PC3 explained 11.0% of the variance and it separated Hiratake from all the other

ur

mushrooms (Figure 1b), as it had the highest concentration of tyramine (0.88 mg/100 g) (Figure 1d). PC4 explained 4.55% of the variance, but no relevant information could be

Jo

obtained from it (Figures 1b and 1d). The HCA dendrogram (Figure 2), built using the Ward’s method, separated into

clusters the different mushroom species. Portobello and Champignon, both from Agaricus bisporus showed no distance between its cluster’s centers, which indicates that their amines’ profiles and contents are very similar., Therefore, it is an efficient tool for the characterization of this specie.

White Shimeji, Salmon, Black Shimeji and Hiratake 15

mushrooms from Pleurotus spp. were also clustered together, but with a higher weighted distance, which indicates that this group presented a great internal variability in amines’ profile and content, as detailed in tables 3 and 4. Even though, the HCA was capable of clustering this Pleurotus species together. Eryngii, another member of the Pleurotus genus, was similar to Shitake, that belongs to Lentinula edodes. This clustering is not surprising, once both mushrooms presented only SPD and AGM at detectable levels for most samples.

But, since the number of Eryngii lots analyzed was too small, further

studies are needed. Therefore, multivariate analysis using bioactive amines’ contents can

ro of

be a potential tool in the discrimination and authenticity determination of mushroom species.

-p

re

4. Conclusions

lP

The ion pair reverse phase liquid chromatographic method followed by post column derivatization with o-phtalaldehyde and fluorescence detection was fit for the analysis of

na

bioactive amines in mushroom. Biologically active amines were detected in fresh edible mushrooms species analyzed, among them spermidine, agmatine, putrescine, cadaverine,

ur

tyramine, tryptamine and phenylethylamine. Spermidine was the prevalent amine followed by agmatine, tryptamine and tyramine.

Putrescine, cadaverine and phenylethylamine

Jo

were found only in a few Pleurotus spp mushrooms. Histamine and serotonin were not detected in any analyzed mushroom.

Spermidine was ubiquitous to the mushrooms,

which is relevant due to its association with growth and health promotion and antioxidant properties.

Agmatine was inherent to Pleurotus spp. and its presence is relevant as

another route for the formation of polyamines.

Furthermore, agmatine has been

associated with beneficial health effects. The levels of spermidine found in mushrooms 16

classify them as high sources of polyamines.

The aromatic vasoconstrictor amines

tyramine, tryptamine and phenylethylamine were also detected in some mushrooms. The levels of cadaverine and putrescine were discrete.

Multivariate analysis based on

bioactive amines contents was effective in showing differences among different types of mushrooms. Thus, the chemometric approach in the analysis of bioactive amines is a promising tool for mushrooms identity.

ro of

Acknowledgements

The authors acknowledge Coordenação de Pessoal de Nível Superior – CAPES, Conselho Nacional de Desenvolvimento Científico e Tecnológico – CNPq (Brasília, DF,

-p

Brazil) and Fundação de Amparo a Pesquisa do Estado de Minas Gerais – FAPEMIG

re

(Belo Horizonte, MG, Brazil) for the financial support.

lP

Conflicts of interest

ur

References

na

The authors declare no conflict of interest.

Alves, J. O., Neto, W. B., Mitsutake, H., Alves, P. S. P., Augusti, R. (2010). Extra virgin

Jo

(EV) and ordinary (ON) olive oils: distinction and detection of adulteration (EV with ON) as determined by direct infusion electrospray ionization mass spectrometry and chemometric approaches. Rapid Communications in Mass Spectrometry, 24, 1875– 1880.

ANPC (Associação Nacional dos Produtores de Cogumelos) (2019). Retrieved May 14, 2019 from: https://www.anpccogumelos.org/cogumelos. 17

AOAC (Association of Official Analytical Chemists) (2012). Official Methods of Analysis. (19th ed.) Maryland: AOAC v. 1, p. 3.1, meth. 931.04. Bandeira, C. M., Evangelista, W. P., Gloria, M. B. A. (2012). Bioactive amines in fresh, canned and dried sweet corn, embryo and endosperm and germinated corn. Food Chemistry, 131, 1355–1359. Cipolla, B. G., Havouis, R., Moulinoux, J. P. (2007). Polyamine contents in current foods: a basis for polyamine reduced diet and a study of its long-term observance and tolerance in prostate carcinoma patients. Amino Acids, 33, 203–212.

ro of

Dadáková, E., Nova, T. P., Kalač, P. (2009). Content of biogenic amines and polyamines in some species of European wild-growing edible mushrooms. European Food Research and Technology, 230, 163–171.

-p

De Cabo, R., & Navas, P. (2016). Spermidine to the rescue for an aging heart. Nature

re

Medicine, 22(12), 1389–1390.

Donnini, D., Gargano, M. L., Perini, C., Savino, E., Murat, C., Di Piazza, S., … Zambonelli,

lP

A. (2013). Wild and cultivated mushrooms as a model of sustainable development. Plant Biosystems, 14(1), 226–236.

na

EFSA (European Food Safety Authority) (2011). Scientific Opinion on risk based control of biogenic amine formation in fermented foods. EFSA Journal, 9, 1–93.

ur

Elmastas, M., Isildak, O., Turkekul, I., Temur, N. (2007). Determination of antioxidant activity and antioxidant compounds in wild edible mushrooms. Journal of Food

Jo

Composition and Analysis, 20 (3-4), 337–345.

FAOSTAT. Food and Agriculture Organization of the United Nations. (2019). Retrieved May 14, 2019 from: http://www.fao.org/faostat/en/#data/QC.

Feeney, M. J., Dwyer, J., Hasler-Lewis, C. M., Milner, J. A., Noakes, M., Rowe, S., Wu, D. (2014). Mushrooms and health summit proceedings. The Journal of Nutrition, 144(7), 1128–1136. 18

Granato, D., Calado, V.M.A., Jarvis, B. (2014). Observations on the use of statistical methods in Food Science and Technology. Food Research International, 55, 137–149. Granato, D., Putnik, P., Kovačevič, D. B., Santos, J. S., Calado V., Rocha, R. S., Pomerantsev, A. (2018a). Trends in chemometrics: food authentication, microbiology, and effects of processing. Comprehensive Reviews in Food Science and Food Safety, 17, 663–673. Granato, D., Santos, J. S., Escher, G. B., Ferreira, B. L., Maggio, R. M. (2018b). Use of principal component analysis (PCA) and hierarchical cluster analysis (HCA) for

ro of

multivariate association between bioactive compounds and functional properties in foods: A critical perspective. Trends in Food Science & Technology, 72, 83–90.

Handa, A. K., Fatima, T., Mattoo, A. K. (2018). Polyamines: bio-molecules with diverse

-p

functions in plant and human health and disease. Frontiers in Chemistry, 6, 1–18.

re

Janjušević, L., Karaman, M., Šibul, F., Tommonaro, G., Iodice, C., Jakovljević, D., Pejin, B. (2017). The lignicolous fungus Trametes versicolor (L.) Lloyd (1920): a promising

lP

natural source of antiradical and AChE inhibitory agents. Journal of Enzyme Inhibition and Medicinal Chemistry, 32, 355–362.

na

Kalač, P. (2013). Chemical composition and nutritional value of European species of wild growing mushrooms: A review. Food Chemistry, 113, 9–16.

ur

Kalač, P. (2014). Health effects and occurrence of dietary polyamines: a review for the period 2005-mid 2013. Food Chemistry, 161, 27–39. Laube, G., & Bernstein, H. (2017). Agmatine: multifunctional arginine metabolite and

Jo

magic bullet in clinical neuroscience? Biochemical Journal, 474, 2619–2640.

Manninen, H., Rotola-Pukkila, M., Aisala, H., Hopia, A., Laaksonen, T. (2018). Free amino acids and 5’-nucleotides in Finnish forest mushrooms. Food Chemistry, 247, 23–28.

Nishibori, N., Fujihara, S., Akatuki, T. (2007). Amounts of polyamines in foods in Japan and intake by Japanese. Food Chemistry, 100, 491–497.

19

Okamoto, A., Sugi, E., Koizumi, Y., Yanagida, F., Udaka, S. (1997). Polyamine content of ordinary

foodstuffs

and

various

fermented

foods.

Bioscience

Biotechnology

Biochemistry, 61, 1582–1584. Önal, C. (2013). A review of the liquid chromatographic methods for the determination of biogenic amines in foods. Food Chemistry, 138(1), 509–515. Otto, M. (2017). Chemometrics – Statistics and Computer Application in Analytical Chemistry. (3rd ed.) Weinheim, Germany: Wiley-VCH. Pop, R. M., Puia, I. C., Puia, A., Chedea, V. S., Leopold, N., Bocsan, I. C., Buzoianu, A. D.

ro of

(2018). Characterization of Trametes versicolor: Medicinal mushroom with important health benefits. Notulae Botanicae Horti Agrobotanici Cluj-Napoca, 46(2), 343-349. Ramani, D., De Bandt, J. P., Cynober, L. (2014). Aliphatic polyamines in physiology and

-p

diseases. Clinical Nutrition, 33(1), 14–22.

Rider, J. E., Hacker, A., Mackintosh, C. A., Pegg, A. E., Woster, P. M., Casero, R. A.

re

(2007). Spermine and spermidine mediate protection against oxidative damage caused by hydrogen peroxide. Amino Acids, 33(2), 231–240.

lP

Rogers, A. C., McDermott, F. D., Mohan, H. M., O’Connell, P. R., Winter, D. C., Baird, A. W. (2015). The effects of polyamines on human colonic mucosal function. European

na

Journal of Pharmacology, 764, 157–163.

Roupas, P., Keogh, J., Noakes, M., Margetts, C., Taylor, P. (2012). The role of edible

709.

ur

mushrooms in health: evaluation of the evidence. Journal of Functional Foods, 4, 687–

Jo

Sharma, S., Kumar, P., Deshmukh, R. (2018). Neuroprotective potential of spermidine against rotenone induced Parkinson’s disease in rats. Neurochemistry International, 116, 104–111.

Silva, A. C., & Jorge, N. (2014). Influence of Lentinus edodes and Agaricus blazei extracts on the prevention of oxidation and retention of tocopherols in soybean oil in an

20

accelerated storage test. Journal of Food Science and Technology, 51 (6), 1208– 1212. Thompson, M., Ellison, S.L.R., Wood, R. (2002). Harmonized guidelines for single laboratory validation of methods of analysis (IUPAC Technical Report). Pure and Applied Chemistry, 74, 835–855. Toro-Funes, N., Bosch-Fusté, J., Veciana-Nogués, M. T., Izquierdo-Pulido, M., VidalCarou, M. C. (2013). In vitro antioxidant activity of dietary polyamines. Food Research International, 51, 141–147.

ro of

Wang, L. C., Thomas, B. W., Warner, K., Wolf, W. J., Kwolek, W. E. (1975). Apparent odor thresholds of polyamines in water and 2% soybean flour dispersions. Journal of Food Science, 40, 274–276.

-p

Yamamoto, S., Itano, H., Kataoka, H., Makita, M. (1982). Gas-liquid chromatographic

re

method for analysis of di- and polyamines in foods. Journal of Agriculture and Food Chemistry, 30, 435–439.

lP

Yang, J. H., Linb, H. C., Mau, J. L. (2002). Antioxidant properties of several commercial mushrooms. Food Chemistry, 77, 229–235.

na

Yen, G. C. (1992). Effects of heat treatment and storage temperature on the biogenic amine contents of straw mushroom (Volvariella volvacea). Journal of Science Food Agriculture,

58,

59–61.

Jo

ur

and

21

oo

f

List of Figures

Fig. 1. Scatter plot [PC1 and PC2 (a), PC3 and PC4 (b)] and PC loadings [PC1-PC2 (c), and PC3-PC4 (d)] obtained for the mean levels

Jo ur

na l

Pr

e-

pr

of bioactive amines of each edible mushroom by Principal Component Analysis.

22

Jo ur

na l

Pr

e-

pr

oo

f

Fig. 2. Dendrogram obtained by Classical Cluster Analysis for the mean of bioactive amines for each edible mushroom.

23

Table 1.

Specie/Mushroom

Other

oo

f

Types of edible mushroom included in this study and their respective species, common names and illustrative photograph. common

Picture

Agaricus bisporus Champignon (A)

Button, Paris, Table or

Portobello (B)

e-

cultivated mushroom Field mushroom

Oat mushroom

Pleurotus spp. Oyster mushroom

White Shimeji (E)

Florida mushroom

Hiratake (F) Salmon (G)

Pink oyster mushroom

Jo ur

Eryngii (H)

na l

Black Shimeji (D)

Pr

Lentinula edodes Shiitake (C)

pr

names

24

Table 2.

oo

f

Fitness parameters (linearity, accuracy, precision and limits of detection and quantification) during method validation for the analysis of bioactive amines in mushrooms. Calibration curves

Accuracy

Slope

Intercept

R2

Spermidine

3.41x105

-22669

0.9993

Agmatine

2.85x105

6009

0.9995

Putrescine

4.50x105

55661

Cadaverine

5.41x105

45657

Tyramine

2.02x105

-29496

Phenylethylamine

3.26x105

Tryptamine Histamine

Limits (mg/100 g)

CVr

CVR

LOD

LOQ

104.4

5.7

7.8

0.03

0.10

99.5

4.4

7.2

0.03

0.10

0.9992

100.7

6.5

8.9

0.03

0.10

0.9995

94.9

6.3

8.7

0.03

0.10

0.9966

101.3

5.9

7.9

0.03

0.10

20243

0.9990

101.3

4.7

6.7

0.03

0.11

2.42x105

115250

0.9990

87.2

5.9

9.5

0.03

0.11

2.99x105

-13880

0.9997

101.4

4.7

7.3

0.03

0.10

2.51x105

-12062

0.9975

70.7

5.8

15.3

0.03

0.10

na l

Pr

e-

(%)

Jo ur

Serotonin

Precision (%)

pr

Analytes

CVr = intraday coeficient of variation - repeatability; CVR = interday coeficient of reproducibility.

Table 3. 25

Occurrence and total levels of free bioactive amines in different types of edible mushroom.

f

Total

Mushroom (n) SPD

AGM

PUT

100

Portobello (13)

100

7.7

e-

Champignon (13)

TRM

PHM

(mg/100 g) 9.97±3.75c

23.1

8.43±2.00c

7.1

7.77±3.70c

21.4

20.3±4.57b

100

7.1

Black shimeji (14)

100

100

White shimeji (13)

100

100

53.8

23.1

30.8

100

84.6

20.3±5.72b

Hiratake (12)

100

100

33.3

16.7

50

16.7

8.3

23.9±4.34b

Salmon (14)

100

100

57.1

57.1

14.3

92.9

21.4

36.6±8.49a

Eryngii (2)

100

100

na l

Pleurotus spp.

7.1

levels

Pr

Lentinus edodes Shiitake (14)

TYM

pr

Agaricus bisporus

CAD

oo

Occurrence of amines* (%)

10.2±1.73c

Jo ur

(n) = number of mushroom lots. * SPD= spermidine; AGM= agmatine; PUT= putrescine; CAD= cadaverine; TYM= tyramine; TRM= tryptamine; PHM= phenylethylamine. Histamine and serotonin were not detected in any mushroom. Total levels of amines with different letters in the same column are significantly different (Tukey test, p≤0.05).

26

8.98

Phenylethylamine

nd (0.00)b nd (0.00)b

nd-0.10 (0.01)a nd (0.00) a

nd-0.77 (0.13)c nd (0.00)c

nd (0.00)b nd (0.00)b

nd (0.00)b

nd (0.00)b

nd-0.28 (0.02)a

nd-0.63 (0.05)c

nd (0.00)b

nd (0.00)b nd-4.53 (0.93)ab nd-3.74 (0.62)b nd-10.4 (2.48)a nd (0.00)

nd (0.00)b nd-0.83 (0.13)ab nd-2.16 (0.30)ab nd-2.72 (0.59)a nd (0.00)b

nd (0.00)a nd-0.70 (0.14)a nd-5.98 (0.88)a nd-2.09 (0.29)a nd (0.00)a

nd-0.63 (0.08)c 0.36-2.73 (1.37)a nd-1.23 (0.18)c 0.32-1.85 (0.82)b nd (0.00)

nd (0.00)b nd-1.91 (0.84)a nd-0.64 (0.05)b nd-0.39 (0.06)b nd (0.00)b

4.41-16.5 (9.83)ab 6.24-12.6 (9.72)ab

nd (0.00)c nd (0.00)c

nd (0.00)b nd (0.00)b

nd-6.86 (0.49)c

5.18-12.6 (6.26)b

Pleurotus spp. Black Shimeji

9.40

White Shimeji

9.66

Hiratake

10.14

Salmon

9.66

6.90-15.7 (12.4)a 3.23-11.5 (7.00)b 4.13-14.2 (9.21)ab 5.68-17.2 (11.5)a 7.13-8.62 (7.87)ab

1.41-13.4 (7.77)b 2.78-13.7 (9.91)b 6.15-15.9 (12.7)b 12.6-34.1 (20.9)a 1.87-2.84 (2.36)c

Jo ur

na l

9.54

11.57

Tryptamine

Putrescine

L. edodes Shitake

Eryngii

Tyramine

Agmatine

Cadaverine

pr

Portobello

9.68

Spermidine

e-

A. bisporus Champignon

Range (mean levels) of amines (mg/100 g)

Pr

Mushrooms

Dry matter (g/100 g)

oo

Levels of free bioactive amines in different types of fresh edible mushrooms.

f

Table 4.

Mean values were calculated using zero as nd (not detected, < 0.03 mg/100 g). Mean values with different letters in the same column are significantly different (Tukey test, p≤0.05).

27

×

Report "Investigation of biologically active amines in some selected edible mushrooms"

Your name

Email

Reason -Select Reason- Pornographic Defamatory Illegal/Unlawful Spam Other Terms Of Service Violation File a copyright complaint

Description

Close Send

Our partners will collect data and use cookies for ad personalization and measurement. Learn how we and our ad partner Google, collect and use data. Agree & close