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Biochemical constituents of palmwine

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Abstract

Biochemical constituents, which include sugars, protein, amino acid, lipid, alcohol, mineral and trace elements of palmwine, a sap obtained from Elaeis guineensis, Jacq (oil palm), are reported. Mean sugar contents ranged between 0.10 in maltose and 8.74 mg/100ml in sucrose. Values of 39.03, 59.63, and 62.65 mg/100ml are reported for protein, free amino acids, and lipids, respectively. Ethanol content was 3.40g/ 100ml. Magnesium, P, and Zn were the most abundant elements, while Pb, Cd, and Co were detected at nontoxic levels. Public health implications of palmwine consumption briefly were addressed. As palmwine is becoming an important source of revenue for the rural poor and as consumer demand increases, it is concluded that the nutritional and economic potential of palmwine should be further explored and given more research attention.
BIOCHEMICAL CONSTITUENTS OF
PALMWINE
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Ecology of Food and Nutrition, 42:255–264
Copyright © 2003 Taylor & Francis
ISSN: 0367-0244 print / 1534-5237 online
DOI: 10.1080/03670240390226222
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Address correspondence to I. E. Ezeagu, Medical Biochemistry Department,
College of Medicine, University of Nigeria, Enugu Campus, Nigeria. E-mail:
ikezeagu@yahoo.com.uk
BIOCHEMICAL CONSTITUENTS OF
PALMWINE
I. E. EZEAGU AND M. A. FAFUNSO
Biochemistry Department, University of Ibadan, Ibadan, Nigeria
F. E. EJEZIE
Medical Biochemistry Department, College of Medicine, Univer-
sity of Nigeria, Enugu Campus, Nigeria
(Received xxx; accepted xxx)
Biochemical constituents, which includes sugars, protein, amino acid, lipid, alcohol,
mineral and trace elements of palmwine, a sap obtained from Elaeis guineensis,
Jacq (oil palm), are reported. Mean sugar contents ranged between 0.10 in maltose
and 8.74 mg/100ml in sucrose. Values of 39.03, 59.63, and 62.65 mg/100ml are
reported for protein, free amino acids, and lipids, respectively. Ethanol content was
3.40g/100ml. Magnesium, P, and Zn were the most abundant elements, while Pb,
Cd, and Co were detected at nontoxic levels. Public health implications of palmwine
consumption briefly were accessed. As palmwine is becoming an important source
of revenue for the rural poor and as consumer demand increases, it is concluded
that the nutritional and economic potential of palmwine should be further explored
and given more research attention.
Keywords: Palmwine, Elaeis guineensis, Biochemical Constituents, Heavy Metals,
Sugars, Public Health
Palm wine is the fermented palm sap obtained by tapping of palms
(family Palmea). Two sources of palmwine in Nigeria are the fer-
256 I. E. EZEAGU ET AL.
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mented sap of oil palm tree (Elaeis guineensis, Jacq) and the Raphia
palm tree (Raphia hookeri, Mann and Wendl). Raphia palms in-
habit swampy regions or areas of wet soil. E. guineensis, otherwise
known as oil palm is the specie of Elaeis genus found in Nigeria. It is
widely distributed in West and Central Africa. Methods of tapping
include tapping at the base of male flower bud (in inflorescence tap-
ping) or at the base of the terminal bud (in stem tapping) (Essiamah
1993; Tuley, 1965a, b). Intensive tapping often results in termination
of growth and death of palm trees. In other parts of the world, like
Philippians, Tunisia, Algeria, and Libya, equivalents of palmwine
are produced by palm trees (Dowson, 1953).
In Nigeria, palmwine has been associated with peasant life be-
cause it is cheaper and produced in the rural areas (Uzogara et al.,
1990). However, as the cost of brewed alcoholic beverages rises, the
trend is changing and demand for palmwine has risen even among
the urban dwellers. The palmwine industry is of considerable eco-
nomic and nutritional importance in West Africa. Palmwine is fast
becoming an important source of revenue for the rural poor, but has
received very little scientific attention.
Fresh palmwine is very sweet and refreshing because of the
presence of sucrose, but within 24 hours the concentration of su-
crose falls to less than 50% the initial amounts (Bassir, 1962). Mi-
croorganisms contaminate the palm sap and convert the sap into
palmwine by a fermentation process. Fermentation virtually ends
when the pH falls to 4.0; this the whole process lasts about 48
hours (Bassir, 1962). Seasonal variations in palmwine constitu-
ents and factors that influence the constituents (including season,
type of soil, time of tapping, and process of storage occur) (Faparusi
and Bassir, 1972a, b). In the light of the growing economic im-
portance of palmwine in Nigeria, is useful to investigate further
the composition and possible nutritional significance of palmwine
obtained within a city environment.
MATERIALS AND METHODS
Palmwine samples were collected fresh (3 to 6 hours old) and were
tapped and pooled from about 40 palm trees around the Ibadan
University campus in the month of March. Samples were collected
BIOCHEMICAL CONSTITUENTS OF PALMWINE 257
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on four different days with the help of a local palmwine tapper and
labelled S1, S2, S3, and S4.
Pretreatment of Samples
Prior to sugar estimation 100 ml aliquots of the palmwine samples
were desalted by first passing through a cation exchange resin—
Amberlite 1R 120 (H+ form)—and amino acids are eluted with 2N
NaOH using Ninihydrin test to indicate complete elution (Redfield,
1953). The residue was dissolved in water and passed through a col-
umn of anion exchange—Amberlite IRA 400 (OH- form). The de-
salted palmwine was then concentrated in vacuo from about 5:1
volume.
Estimation of Sugars
For each sample, 5µl of were spotted in triplicates along side a mix-
ture of each standard sugar solution on separate Whatman No. 1
chromatography papers. The sugars were separated by means of one-
way descending paper chromatography method as described by
McFarren et al. (1951) and Durso and Paulson (1958) using
ethylacetate-pyridine- water (12:4:4 v/v) as the separating solvent and
allowed to run for 16 hours. Ammonical silver nitrate solution was
used as the locating reagent as described by Travalyan (1950). The
Rf values were calculated and recorded. Areas containing the spots
were marked out and corresponding areas on each of the remaining
unspread duplicates were removed and eluted by direct dipping of
each sugar regions in 5 ml distilled water contained in boiling tubes
for 4 hours. The respective sugar concentrations were determined
by the phenol-sulphuric method and were read off their standard
curves by extrapolation.
Total Nitrogen, Free Amino Acid, and Protein Determination
10ml aliquots of samples were digested and total nitrogen determined
by microKjeldahl method. To determine true protein and free amino
acid contents, HgCl2 (10 mg) were added to 10 ml aliquots of samples
to precipitate proteins and was centrifuged at 3,000rpm for 10 min-
utes. Nitrogen in both the precipitate and supernatant were deter-
258 I. E. EZEAGU ET AL.
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mined by microKjeldahl method as protein and free amino acid N,
respectively.
Determination of Total Lipids
Total lipid was estimated by the method of Folch et al. (1957). Miner-
als were estimated by the wet digestion method (AOAC, 1984) using
an atomic absorption spectrophotometer (Perkin Elmer 305B). Phos-
phorus was determined colorimetrically by phospho-vanado-molyb-
date method (AOAC, 1984). Specific gravity (SG) and pH of samples
were determined using a hydrometer and a pH meter, respectively.
Alcohol content was determined by distillation of a 100 ml sample
until 90–95 ml of distillate was collected at 20oC. The SG of the
distillate was determined and the percentage of alcohol by volume
was read-off a conversion table of SG of alcohol (AOAC, 1984).
RESULTS AND DISCUSSION
Results of alcohol, pH, and SG are reported in Table 1. Alcohol
content ranged between 2.7 and 5.2 gm/100ml, similar to the 5%
ethanol level reported by Osin et al. (1991). Fresh palm sap usually
contains no alcohol but levels could rise to 4.5–5.2gm/100 ml after
72 hours and may fall slightly after the fifth day of storage due prob-
ably to oxidation of alcohol by invading microorganisms (Bassir, 1962;
Faparusi and Bassir, 1972b). A mean SG value of 1.0 shows that the
palm sap contains mainly water. The pH was between 7.2–7.4.
Faparusi and Bassir (1972b) observed that pH fell sharply after the
first two days of storage and also that acidity was due to production
of tartaric, acetic, and lactic acids.
TABLE I
Specific Gravity, pH, and Alcohol Contents of Palmwine
Samples S1 S2 S3 S4 Mean±SD
SG 0.99 0.99 1.0 0.99 0.99±0.01
pH 7.2 7.4 7.3 7.4 7.32±0.10
Alcohol 2.8 3.1 2.7 5.2 3.05±1.08
(g/100ml)
BIOCHEMICAL CONSTITUENTS OF PALMWINE 259
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Plates I and II of Table 2 shows the qualitative analysis of sugar
and the corresponding Rf values as measured from the chromato-
grams. Presence of raffinose (Rf = 0.163), sucrose (Rf = 0.326), and
glucose (Rf = 0.372) was indicated on Plate I while on Plate II the
presence of maltose, glucose (Rf = 0.419), and fructose (Rf = 0.442)
were obvious. Other reference sugars—xylose, arabinose, and lac-
tose—either were absent or present in undetectable amounts in the
samples. The quantitative estimates of sugars in the palmwine samples
are shown in Table 3. Concentration of sucrose seems very low (2.50–
2.83 mg/100 ml), and hence was not quite clear on the chromato-
gram. The low content of sucrose can be explained by the fact that
sucrose is preferentially utilized in the fermentation medium by yeasts
(Bavisotto et al., 1958). Leibowitz and Hestrin (1945) showed that a
brewery yeast strain removed sugars from wort in the following or-
der: sucrose, monosaccharides, maltose, and maltotriose. Sucrose
also can be hydrolyzed by invertase to glucose and fructose. The two
sugars are then broken down by enzymes to produce ethanol. Inver-
tase has been identified in appreciably high amounts in palm sap
and could be responsible for the absence or rapid decrease in su-
crose concentration (Visser and Bassir, 1969). Sucrose level there-
fore would be dependent on the degree of fermentation that has taken
place prior to analysis
Maltose levels were low which bears on the use of fresh samples
in this study. Maltose could only be identified in palm sap after the
TABLE II
Rf of Reference Sugars on Paper Chromatogram
Reference Sugar Plate I Plate II
Fructose 0.465 0.442
Xylose 0.512 NS
Raff inose 0.163 0.186
Maltose 0.302 0.302
Sucrose 0.326 NS
Glucose 0.372 0.419
Lactose NS 0.233
Arabinose NS 0.465
Rf = distance moved by solute from origin
distance moved by solvent from origin
NS = reference not spotted
260 I. E. EZEAGU ET AL.
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first day of storage (Faparusi and Bassir, 1972b). Mean glucose, fruc-
tose, raffinose, and maltose levels are 0.65, 0.90, 0.32, and 0.10, re-
spectively. Total sugar levels ranged between 4.36–5.26 mg/100 ml.
Faparusi (1966) also reported the presence of glucuronic acid and
noted that glucuronic usually is identified only after the first day and
concentrations remain constant on storage. However, variations in
sugar contents of palm sap usually occur depending on the period of
tapping and storage due to microbial activities. Microorganisms re-
ported in palmwine include Bacillus, Streptococcus, Saccharomy-
ces, Schizosaccharomyces, Pischia, Leuconostoc, Micrococcus,
Serratia, Aerobacter, Pseudomonas, Cornybacterium, Asppergillus,
and Candida (Bassir, 1962, Omobuwajo, et al., 1987)
Total nitrogen, protein, free amino acids, and lipid contents are
shown in Table 4. The total nitrogen and free amino acids varied
between 85.12–119.7 and 50.5–66.5 mg/100 ml, respectively. Mean
protein content was 39.03 mg/100 ml. The proteins may have been
produced by the plant and are being translocated in the palm sap.
Occurrence of high protein levels, coupled with the oil and high
sugar contents, could be of nutritional significance. The amino ac-
ids are probably of plant origin too, but microorganisms also may
TABLE III
Sugar Components of Palmwine (mg/100 ml)
SAMPLE S1 S2 S3 S4 Mean±SD
Fructose 1.05 0.80 0.80 0.95 0.9±0.12
Glucose 0.75 0.50 0.75 0.60 0.65±0.11
Sucrose 2.83 2.63 2.50 2.70 2.67±0.14
Maltose 0.13 0.09 0.09 0.10 0.10±0.02
Raff inose 0.50 0.30 0.37 0.10 0.32±0.17
Total Sugar 5.26 4.32 4.51 4.45 4.64±0.42
TABLE IV
Protein, Free Amino Acids, and Total Lipids in Palmwine (mg/ 100ml)
S1 S2 S3 S4 Mean±SD
Total N 101.1 85.12 119.7 93.1 99.76±14.81
Free Amino Acid 66.5 50.5 66.5 55.0 59.63±8.15
Protein 34.6 34.6 47.9 39.0 39.03±6.27
Lipids 83.0 55.0 54.9 57.7 62.65±11.80
BIOCHEMICAL CONSTITUENTS OF PALMWINE 261
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contribute to amino acid production (Sjoblohm, 1965). Mean lipid
content was 62.65 mg/ml. E. guineensis is a well-known economic
source of edible red palm oil, which is very rich in palmitic acid, a
precursor of vitamin A.
Macro and micro mineral element contents are shown in Table
5. Magnesium and P (32.0 and 59.75mg/L, respectively) were the
most abundant minerals. Cadmium, Pb, and Co occurred in very
low levels (<0.1 ppm), though levels may vary widely with location.
Some elements such as Pb, Cd, and Zn could act as cumulative poi-
sons if long-term, low-level exposure and possible build-up to thresh-
old levels occur (Onianwa et al., 2001; William, 1977). However such
a situation usually is averted by the homeostatic mechanism of the
body, except in disease conditions or where the exposure is too high
for the body to handle. Palmwine usually is taken occasionally, al-
though an average consumer may take up to 2.5 L at a time. Based
on a 2.5 L daily consumption of palmwine, the potential intake of
heavy metals was compared to the normal acceptable daily intake
(ADI) (Table 6). Only Co and Cu (1.2–2.5 and 6.25–12.5 mg/2.5L,
respectively) intake exceeded recommended normal ADI levels, even
though these levels were lower than the toxic levels. Such occasional
consumption of palmwine therefore is not considered hazardous.
Other constituents variously reported in palmwine include ni-
trates (18.16–91.26 mg/L), nitrite (0.00–2.81mg/L) dimethylamine
(4.06–28.36 mg/L), (Bassir and Maduagwu, 1978; Ezeagu, 1995) and
tyramine (11.27 mg/100 ml) (Uzogara et al., 1987). The public health
TABLE V
Mineral Composition of Palmwine (mg/L)
SAMPLE S1 S2 S3 S4 Mean±SD
Cadmium <0.1 0.1 0.1 0.1 0.10±0.0
Cobalt 1.0 1.0 0.5 1.0 0.88±0.25
Copper 2.5 3.8 3.8 5.0 3.78±1.02
Iron 3.0 2.0 2.5 3.0 2.63±0.48
Magnesium 28.0 40.0 30.0 30.0 32.00±5.42
Manganese 1.25 1.25 1.30 1.25 1.26±0.03
Phosphorus 60.0 59.0 60.0 60.0 59.75±0.50
Lead <0.10 0.10 0.10 0.10 0.10±0.00
Zinc 2.0 1.8 2.0 2.0 1.95±0.10
Calcium 0.10 0.4 0.8 0.6 0.48±0.30
262 I. E. EZEAGU ET AL.
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implications of these factors have been reviewed in the literature,
though their levels in palmwine are of little or no hazard (Uzogara et
al., 1987; Van Maanen et al., 1994; Walker, 1990). According to Osim
et al. (1991), fresh palmwine is not ulcerrogenic as might be thought
despite its content of 5% ethanol.
CONCLUSIONS
As palmwine is becoming an important source of revenue for the
rural poor, and as consumer demand increases, the nutritional and
economic potential should be further explored. Expanding its culti-
vation from family farms to plantation scale will ensure a sustainable
and commercial exploitation, both at local and national levels.
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Bavisotto V.S, L.A. Roch, and E.J. Petrusek (1958). Chromatographic evaluation
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Heavy Metal Levels in a Daily Palmwine Intake
Element Sample 2.5L/Day ADIaToxic Levels
Mean intake (mg/day) (mg/day)
(mg/L) (mg)
Cd 0.10 0.25 0.06 3.0
Co 0.88 2.20 0.0002 500
Cu 3.78 9.45 2–5 250–500
Fe 2.63 6.58 12–15 760
Mn 1.26 3.15 3–9
Pb 0.10 0.25 0.30–0.40 1000
Zn 1.95 4.88 10–15 509(rat)
aAverage man (70 kg body weight) (Bowen, 1966).
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... The antibacterial activity of amoxicillin /clavulanic acid against all the test organisms was reduced (increase in MIC) in the presence of the freshly tapped sap from oil palm. The MIC of coamoxiclav increased against S. aureus (8-fold), Doddipalla et al. (2022) and Ezeagu et al. (2010) P. mirabilis (510-fold), S. paratyphi A (32-fold), S. paratyphi B (32-fold), P. aeruginosa (4-fold), E. coli (>1041-fold) and K. pneumoniae (16-fold). There was no change in the MIC of cefixime against P. aeruginosa and K. pneumoniae. ...
... LC-MS screening of the fresh sap from oil palm principally showed the presence of monosaccharides (like glucose), disaccharides (including cellobiose, maltose, and sucrose), and ascorbic acid (Figure 1 and Table 2). The presence of these compounds has been previously reported in the sap of oil palm tree (Doddipalla et al., 2022;Ezeagu et al., 2010). The sweet taste of the product could thus be attributed to the presence of the free sugars. ...
... These organisms have also been reported to originate from several sources, which include tapping equipment, containers, the environment, etc [6]. Generally, both brands of palm wine have several nutritional, medical, religious and social uses which have been reported [7] to have increasingly enhanced the demand for this natural product. Although attempts has been made towards the preservation and shelf-life extension of palm wine through bottling, use of chemical additives and addition of plant extracts have greatly affected the organoleptic quality of the product [8,4]. ...
... It is produced and consumed in very large quantities in the southeastern Nigeria. The wine is rich in such nutrients as sugars, amino acids, proteins, vitamins and minerals [7]. These make this wine a veritable medium for the growth of a consortium of microorganisms, whose growth in turn, change the physicochemical conditions of the wine, giving rise to competition and successions of organisms. ...
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The enzymatic potentials of Lactic acid bacteria (LAB) isolated from fresh palm wine in Ikwuano L. G. A namely Umuariaga, Amawom, Oboro and some markets around Umuahia North and South in Abia State were evaluated. Fresh palm wine samples were collected from the tappers between 6 and 9am using sterile labeled bottles. The bottled were packed in ice packed container and quickly transported to the laboratory for analyses. The samples were serially diluted and inoculated by spread plate method in duplicates onto De Man Rogosa Sharpe (MRS) Agar and incubated at 35°C for 48 hrs for the isolation of LAB. Six LAB isolates namely Lactobacillus plantarum, Lactobacillus pentosus, Lactobacillus brevis, Lactobacillus fermentum, Lactococcus spp and Leuconostoc spp were recovered from fresh palm. Result showed that all the isolates gave positive amylase, protease and lipase activities. The optimum temperature for enzymatic activity was 50°C for the three enzymes while the optimum pH was in the range 5.0. At the optimum temperature, amylase activity was 9% at pH 5.0, lipase activity was 5.33% and protease activity was 6.14%. The LAB isolates also demonstrated resistance to amoxyl and tetracycline antibiotics. It was concluded that palm wine is a good source of different species of Lactic Acid Bacteria and as such has the potential for increases utility value as raw materials for industries in sourcing for different enzymes that can work at thermophilic temperatures typical of industrial setting.
... Palm wine is a natural drink that is typically used to refresh after meals; therefore, it is important to consider its nutritional content in that context. According to Ezeagu et al. (2003), the sap of the palm tree is considered safe and contains probiotic-containing Saccharomyces yeast species and helpful lactic acid bacteria. It also contains sugar, protein, fat, and mineral matter. ...
... As the level of yeast cell increases, the level of sugar reduces. In the first days of tapping the palm wine is very sugary and does not contain substantial concentration of alcohol (Ezeagu et al., 2003;Amoa-Awua et al., 2007;Karamoko et al., 2012;Santiago-Urbina et al., 2013). As storage time increases, the sugar level reduces because the yeast is fermenting the sugar alcohol. ...
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This research work was carried out investigate the microbiological and biochemical changes taking place in stored palm wine. Microbiological examination of the stored samples revealed that as the storage time increases, the fugal (yeast) cell increases from 2.3 x 102 to 4.7 x 105 cfu/ml. Also, analysis on the sugar level showed that fresh palm wine which forms day 1 sample has sugar level of 38% which makes it sugary. As fermentation process occurs, the sugar is been converted to alcohol. Subsequently, the alcohol formed during fermentation is further oxidized to Aldehyde. There is the possibility of formation of carboxylic acid and ketone. All these chemicals are toxic to the body. This work is recommending that as the storage time increases, the number of yeast cell increases. This yeast can be extracted, purified and kept for use or sale. Consumption of palm wine above four (4) days should be discontinued as the consumer is loading his body with alcohol and aldehyde. This long storage time will also lead to the formation of carboxylic acid and ketone which are harmful to the body.
... The growth and invertase activity of the yeast cells is most likely enhanced by the increase in total acidity and reduction in pH which is a result of the organic acids produced, while acetic acid bacteria use the ethanol generated by the yeast as a substrate for the production of acetic acid [8,11,5]. A fresh palm wine sap has a near-neutral pH that drops after tapping, with a sugar content of just about 10-18% w/v [12,5,3,4,13]. A reduction in the pH of palm wine in the course of fermentation has been observed to be due to organic acids produced mostly by the lactic acid bacteria, followed by acetic acid, which is seen as part of the palm wine aroma [5,4]. ...
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... The mineral composition of palm wine varies notably across different species. For instance, Elaeis guineensis palm wine showcases significant concentrations of magnesium and phosphorus [50], along with several other minerals in varying amounts, while Phoenix dactylifera palm wine is especially rich in potassium, in addition to magnesium and phosphorus [51]. Furthermore, the fermentation process in palm wine, facilitated by microbial activities, introduces a range of organic acids and alcohols which are not innately present in the sap. ...
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... At the vegetative stage, the crop is characterized by continuous stem elongation (Adeneyi AA, Akpabio UD, 2011), with an inflorescence which is the sink for the photosynthate which is tapped and referred to as "palm wine". Other parts of the raphia palm tree such as the leaves, roots, branches and seed are also exploited for craft work and traditional medicines (Mphoweh et al., 2009), (Obahiagbon FI, 2009), (Ezeagu et al., 2003) 88 .The inflorescence emerges from the base of the fanlike leaves, and they bear the male and female flowers that will develop to form the fruits and the seeds. ...
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