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Journal homepage: www.fia.usv.ro/fiajournal 

Journal of Faculty of Food Engineering,  
Ştefan cel Mare University of Suceava, Romania  

Volume XIV, Issue 3 - 2015, pag. 245  -  249 

 

 

SEPARATION AND QUANTIFICATION OF BIOGENIC AMINES IN BANANAS BY 

HIGH PERFORMANCE LIQUID CHROMATOGRAPHY 

 

*Veronica TANASA, Daniela MOISE, Maria STANCA 

 

Chemistry and Biochemistry Analysis Laboratory, Institute of Research and Development for Industrialization 

and Marketing of Horticultural Products - HORTING, 1A Intrarea Binelui, 4 District, Bucharest, Romania 

E-mail: vero.tanasa@yahoo.co.uk 

*Corresponding author 

Received July 13th 2015, accepted September 29th 2015 

 
Abstract: This paper reports separation and quantification of biogenic amines in bananas by high 

performance liquid chromatography. Fresh and preserved samples (seven days of storage at 

refrigeration temperature, at room temperature respectively) were analyzed. We obtained the 

separation of all nine biogenic amines (tryptamine, 2-phenylethylamine, putrescine, cadaverine, 

histamine, serotonin, tyramine, spermidine, spermine) with good resolution, including good resolution 

of serotonin and internal standard (1,7diaminoheptane). Mean biogenic amines levels of 10.344 mg/kg 

(tryptamine) to 61.502 mg/kg (putrescine) were found in fresh bananas. After seven days of storage at 

room temperature, 2-phenylethylamine decreases significantly. After seven days of storage at 

refrigeration temperature, tryptamine increases significantly, whereas phenylethylamine decreases 

significantly. 

 

Keywords:2-phenylethylamine, bananas, biogenic amines, cadaverine, histamine, HPLC, putrescine, 

spermidine, spermine, serotonin, tryptamine, tyramine 

 

 

1. Introduction  

 

Biogenic amines are natural low molecular 

weight organic bases of aliphatic, aromatic 

or heterocyclic structure derived from 

decarboxylation of the corresponding 

amino acids. Biogenic amines are present 

in small quantities in some products such 

as fruits, vegetables, milk etc. 

Polyamines such as putrescine, spermidine 

and spermine have been found in all higher 

plants and are involved in important 

physiological processes, such as fruit 

growth and development [1cited by 2]. 

Free biogenic amines in fruits and 

vegetables shape the typical and 

characteristic taste of mature foods and are 

precursors of certain aroma compounds [3 

cited by 4]. 

Biogenic amines can be found in relatively 

large amounts in some fermented/ matured 

foodstuffs. Also biogenic amines can be 

used as indicators of quality and freshness, 

especially for animal raw material and 

food, as food spoilage is accompanied by 

increased content of biogenic amines. 

Biogenic amines exert physiological and 

toxicological effects. Some aromatic 

amines (tyramine, tryptamine, and 2-

phenylethylamine) show a vasoconstrictor 

action while others (histamine and 

serotonin) present a vasodilator effect. 

Despite of low levels of biogenic amines in 

fruits, vegetables and fruit juices [5, 2, 6, 7 

8], quantification of biogenic amines is 

important due to the existence of risk 

consumer categories. These include both 

patients treated with painkillers and drugs 

used for the treatment of stress and 

http://www.fia.usv.ro/fiajournal


Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefancel Mare University - Suceava 

Volume XIV, Issue 3  – 2015 

Veronica TANASA, Daniela MOISE, Maria STANCA,Separation and cuantification of biogenic amines in bananas by 

High Performance Liquid Chromatography, Food and Environment Safety, Volume XIV, Issue 3 – 2015, pag. 245 – 249  

 246 

depression, acting as a blocker of 

monoamine oxidase - the main route of 

biogenic amines detoxification [9], and 

young children and infants, as they are 

early feed with bananas, at 5 - 6 months of 

age. 

Through our research we aimed at the 

separation and quantification of biogenic 

amines in bananas by high performance 

liquid chromatography, since few data are 

reported about this topic [10, 11, 7, 12]. 

 

2. Materials and methods 

 

2.1. Samples 

Bananas were obtained from Romanian 

retail stores and were analyzed fresh and 

after preservation:  seven days of storage at 

refrigeration temperature, respectively at 

room temperature. 

 

2.2. Reagents and standards 

Tryptamine (Trp), 2-phenylethylamine 

(Phe), putrescine (Put) , cadaverine (Cad), 

histamine (His), serotonin (Ser), tyramine 

(Tyr), spermidine (Spd), spermine (Spm) 

and internal standard 1,7 diaminoheptane 

were purchased from Sigma-Aldrich. Stock 

solutions of biogenic amines and internal 

standard (1 mg/ml) were prepared, which 

were kept in the refrigerator for a month. 

These solutions were used for the 

preparation of standard solutions 

(100μg/ml) used for calibration curves. All 

other reagents used were p.a. grade and 

solvents were HPLC grade. 

 

2.3. The analytical procedure has been 

adapted in the Chemistry and Biochemistry 

Laboratory of the HORTING Institute [13] 

following a previously published protocol 

[14]. In our case chromatographic 

separation was performed using a BDS 

Hypersil C18 column (250 * 4.6 mm), 5μ 

particle size (Thermo Electron 

Corporation). The mobile phase consisted 

of ammonium acetate 0.1 mol / l (solvent 

A) and acetonitrile (solvent B). We 

modified the gradient elution program 

proposed by Eerola et al. (1993) [14] 

(Table 1). The mobile phase flow rate was 

of 1ml/min and the column temperature 

was set at 400C. 

Dansylated derivatives of biogenic amines 

were UV detected at 254 nm. The control, 

data acquisition and processing software 

used ChromQuest 4.2. (ThermoFinnigan).  

The results were statistically processed 

with Student test using GraphPad Prism 

(version 5.00, GraphPad Software Inc., San 

Diego, 2007). The differences p <0.05 

were considered significant. 

 

3. Results and Discussion 

 

By modifying the gradient elution 

programme proposed by Eerola et al. 

(1993) [14] we found that resolution of 

serotonin and the internal standard was 

good (Fig. 1). Eerola et al. (1993) [14] 

asserted that the overlapping of serotonin 

and internal standard peaks did not 

interfere with the quantitation because the 

samples investigated contained no 

serotonin or only very small amounts.  

All biogenic amines investigated were 

present in fresh samples, but putrescine 

was predominant (Table 2). Kalac & 

Krausova (2005) [6] and Okamoto (1997) 

[5] reported that the putrescine content was 

predominant in grapefruit and apple, and 

orange respectively. Cirilio et al. (2003) 

[2] reported that the predominant amines in 

green coffee were serotonin and 

putrescine. Putrescine concentration in 

fresh samples (61.502 mg/kg) was 

comparable with the values reported by 

Kalac & Krausova (2005) [6] in grapefruit 

and fresh green pepper (62.1 respectively 

70 mg/kg). Moret et al. (2005) [4] carried 

out a survey on free biogenic amine 

contents in fresh vegetable products and 

reported that putrescine was one of the 

most represented amines. Adao & et al. 

(2005) [11] found a putrescine content 

around 0.75mg/100g in bananas. 



Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefancel Mare University - Suceava 

Volume XIV, Issue 3  – 2015 

Veronica TANASA, Daniela MOISE, Maria STANCA,Separation and cuantification of biogenic amines in bananas by 

High Performance Liquid Chromatography, Food and Environment Safety, Volume XIV, Issue 3 – 2015, pag. 245 – 249  

 247 

 

 
Fig. 1. Chromatographic separation of dansyl derivatives of biogenic amines (2 μg/ml) by gradient elution 

 

 

 

Fig. 2. Chromatogram of biogenic amines extracted from bananas after 7 days of storage at room 

temperature 

 

  

 

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Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefancel Mare University - Suceava 

Volume XIV, Issue 3  – 2015 

Veronica TANASA, Daniela MOISE, Maria STANCA,Separation and cuantification of biogenic amines in bananas by 

High Performance Liquid Chromatography, Food and Environment Safety, Volume XIV, Issue 3 – 2015, pag. 245 – 249  

 248 

 Table 1. 

Gradient elution programme for the separation of biogenic amines in bananas 

 

Table 2.  

Mean biogenic amines levels (mg/kg) in bananas 

 Trp Phe Put Cad His Ser Tyr Spd Spm 

Fresh samples 10.344 

±0.346 

39.42 

±6.091 

61.502 

±16.91 

32.313 

±4.455 

17.991 

±8.71 

25.379 

±5.664 

14.831 

±0.839 

23.303 

±1.199 

19.164 

±3.218 

Samples preserved 7 

days at room 

temperature 

13.759 

±5.652 

12.595 

±5.464 

p<0.05 

51.381 

±27.605 

24.784 

±10.268 

25.271 

±7.355 

25.265 

±19.756 

11.537 

±7.567 

18.154 

±8.125 

6.084 

±3.486 

Samples preserved 7 

days at refrigeration 

temperature 

17.971 

±1.126 

p<0.05 

14.300 

±2.621 

p<0.05 

57.082 

±16.214 

29.859 

±1.199 

28.053 

±1.352 

21.663 

±4.966 

13.978 

±0.207 

22.436 

±5.418 

9.104 

±5.430 

 

Histamine (17.991 mg/kg) was detected in 

fresh samples. The presence of histamine 

was not detected in bananas by Lavizzari 

et al. (2006) [7], but G.G. Mohamed et al. 

(2009) [12] reported histamine levels of 0–

33.10 mg/100g in seven banana samples. 

The tiramine concentrations in fresh 

samples (14.831 mg/kg) were in agreement 

with those values obtained by Udenfriend 

et al. (1959) [10] in bananas (0.7 

mg/100g). 

We detected tryptamine, 2-

phenylethylamine, cadaverine in fresh 

samples, but these biogenic amines have 

not been detected in bananas by Lavizzari 

et al. (2006) [7].  

Mean serotonin levels of 25.379 mg/kg 

were found; lower amounts of serotonin 

were found by Lavizzari et al. (2006) [7], 

Adao& et al. (2005) [11] in bananas and 

Cirilio et al. (2003) [2] in green coffee 

(11.5 mg/kg, around 1.25 mg/100g 

respectively 1.13 mg/100g ). 

Mean spermidine and spermine levels of 

23.303 respectively 19.164 mg/kg were 

found. Adao & et al. (2005) [11] found a 

spermidine content around 1.10 mg/100g 

in bananas and spermine was present at 

very low level. Cirilio et al. (2003) [2] also 

reported prevalence of spermidine to 

spermine in green coffee with mean levels of 

0.60 mg/100g, 0.44 mg/100g respectively.  

Moret et al. (2005) [4] reported prevalence of 

spermidine to spermine in fresh vegetables 

(0.4–4.5 mg/100 g, a maximum of 1.1 

mg/100 g respectively).  

The samples analyzed after seven days at 

room temperature showed significant lower 

2-phenylethylamine content (Table 2) and no 

differences were found for the other biogenic 

amine levels, while putrescine was the most 

represented amine.  

The samples analyzed after seven days of 

refrigeration showed a statistical significant 

higher tryptamine content and a lower 

phenylethylamine content, and no differences 

were found for the others biogenic amine 

levels, while putrescine was  the most 

represented amine (Table 2). 

It is widely known that some aromatic 

biogenic amines (tyramine, tryptamine, 2-

phenylethylamine) exert a vasoconstrictor 

effect, and the quantification of these 

biogenic amines is important, especially due 

to the existence of risk consumer categories. 

With regard to biogenic amines' content 

Time 

(min) 

Gradient 
Flow 

rate 

(ml/min) 

Wavelength 

(nm) 

Column 

pressure 

(bar) 

Column 

temperature 

(ºC) 

Tray 

temperature 

(ºC) 

Sample 

volume 

(µl) 

Ammonium 

acetate (A) 

(%) 

Acetonitrile 

(B) 

(%) 

0.01 40 60 

1.00 254 
minim 

70 
40 7 20 

15 40 60 

20 30 70 

25 5 95 

30 40 60 



Food and Environment Safety - Journal of Faculty of Food Engineering, Ştefancel Mare University - Suceava 

Volume XIV, Issue 3  – 2015 

Veronica TANASA, Daniela MOISE, Maria STANCA,Separation and cuantification of biogenic amines in bananas by 

High Performance Liquid Chromatography, Food and Environment Safety, Volume XIV, Issue 3 – 2015, pag. 245 – 249  

 249 

modification during refrigeration, Moret et 

al. (2005) [4] reported that vegetables 

analyzed after a 3-week refrigeration 

period showed a lower spermidine content 

and minor differences were found for 

concentrations of other amines. Further 

investigation on more samples is required. 

 

4. Conclusion 

 

a) We obtained the separation of all nine 

biogenic amines from bananas by HPLC 

with good resolution, including good 

resolution of serotonin and internal 

standard (1,7 diaminoheptane).  

b) Mean biogenic amines levels of 10.344 

mg/kg (tryptamine) to 61.502 mg/kg 

(putrescine) were found in fresh bananas. 

c) After seven days of bananas storage at 

refrigeration temperature, their content in 

2-phenylethylamin decreased significantly, 

while their tryptamine content increased 

significantly. 

d) After seven days of storage at room 

temperature, 2-phenylethylamine 

decreased significantly in bananas.  

 

5. Acknowledgments 
 

We would like to thank the Romanian Ministry of 

Education and Scientific Research - National 

Authority for Scientific Research (ANCS), for 

supporting this work under the Excellence 

Research Program, CEEX IV Project - Contract 

no. 115/10.08.2006.  
All the authors declare no conflict of interest. 

 

 

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