ARTICLE IN PRESS

JOURNAL OF
FOOD COMPOSITION
AND ANALYSIS
Journal of Food Composition and Analysis 16 (2003) 613–619
www.elsevier.com/locate/jfca

Original Article

Physico-chemical properties and estimation of mineral content

in honey produced from different plants in Northern India
Vikas Nandaa,*, B.C. Sarkara, H.K. Sharmaa, A.S. Bawab
a
Department of Food Technology, Sant Longowal Institute of Engineering and Technology, Longowal, Punjab, India
b
Defense Food Research Laboratory, Mysore, Karnataka, India
Received 26 August 2002; received in revised form 14 February 2003; accepted 14 March 2003

Abstract

The aim was to study the affect of floral origin on the physico-chemical characteristics and the content of
potassium, sodium, zinc, iron, calcium and copper in honey from six different sources viz., Trifolium
alexandrinum L. (berseem clover), Brassica campestris (mustard), Helianthus annuus (sun flower), Eucalyptus
lanceolatus, Citrus flower, Multi-flower and in one commercial sample. Physico-chemical constituents were
examined according to AOAC methods (in: K. Helrich (Ed.), Official Method of Analysis, 15th Edition,
Association of Official Analytical Chemists, Inc., Arlington, VA, USA, 1990) and minerals were determined
by atomic absorption spectrophotometry. Among the sources of honey significant differences in the amount
of mineral substances, moisture content, and total acidity were found (Po0:01). There was no significant
difference in specific gravity due to source (Po0:01). The levels of sodium, potassium, calcium, iron,
copper, zinc varied from 163.3–304.31, 489.52–932.56, 32.6–84.63, 8.86–13.25, 1.74–2.9, 2.5–16.77 mg/kg,
respectively. Trifolium honey showed the highest moisture content and lowest specific gravity however total
acidity and free acid were highest in citrus honey. Optical rotation was levorotatory in all honey types.
r 2003 Elsevier Ltd. All rights reserved.

Keywords: Pollen; Sunflower; Citrus flower; Mustard; Honey and potassium

1. Introduction

The total world production of honey is estimated at 1 170 000 tons and China, the largest
producer, exports 70 000 tons of honey compared to 1000 tons exported by India (Wakhle, 1998).
In India, the consumption of honey as food is very low, 8.4 g/year/person compared to 120–1800 g
in other countries, due to food habits, high cost and utilization of a major quantity for medical

*Corresponding author: S/o Karam Chand Nanda, 6073 Modren Housing Complex, Manimajra, Chandigarh (UP),
India.
E-mail address: vikasnanda2003@yahoo.co.in (V. Nanda).

0889-1575/03/$ - see front matter r 2003 Elsevier Ltd. All rights reserved.
doi:10.1016/S0889-1575(03)00062-0
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614 V. Nanda et al. / Journal of Food Composition and Analysis 16 (2003) 613–619

purposes (Wakhle, 1998). The major sugars present are glucose and fructose followed by lower
concentrations of sucrose and maltose (Siddiqui & Furgula, 1976). It also contains vitamins such
as vitB1, B2, C and nicotinic acid. About 18 organic acids have been detected in honey. The
mineral content and trace element in honey samples could give an indication of the geographical
origin of honey (Rodriguez-Otero, Paserio, Simal, & Cepeda, 1992). Pollen frequencies of eight
honey samples from Malaysia gave consistent results and reflected the major crops or vegetation
types where the hives were situated and therefore it was possible to distinguish honey from areas
where coconut or star fruit is the predominant crop (Abdullah, 1986). Physical and chemical
properties of different types of honey have been reported by many scientist (Singh, Singh, Bawa,
& Sekhon, 1988; Sancho, Muniategui, Huidobro, & Simal, 1992; Gupta, Kaushik, & Joshi, 1992;
Rodriguez-Otero, Paserio, Simal, & Cepeda, 1994). Though a large number of honey types
varying in composition, cost, shelf life and packing are marketed, it is rather surprising that no
data is available on the mineral composition of various honey types produced in India. Therefore,
the present study was undertaken to ascertain variation in physico-chemical properties and
mineral composition of the honey types in Northern India.

2. Materials and methods

The samples of raw honey from six different sources viz. Trifolium alexandrinum L. (berseem
clover), Brassica campestris (mustard), Helianthus annuus (sun flower), Eucalyptus lanceolatus,
Citrus flower and Multi-flower were harvested from the hives, which were located in the area
where above-mentioned crops constitute the major flora, for the collection of nectar. A
commercial sample of honey was also procured from the local market .The botanical origin of the
samples of honey was based on the pollen spectrum (45% and above), which is the ratio of the
frequency of each pollen type in the honey (Louveaux, Maurizio, & Vorwohl, 1978). The analysis
was based on the principle that microscopic elements were concentrated by centrifuging the honey
dissolved in water, examining the sediments and evaluating them under the microscope after
acetolysis. The method followed for pollen analysis was described by Louveaux et al. (1978).
Briefly, a sub-sample of 10 g of honey was dissolved in 20 ml of warm distilled water (around
40
C) and centrifuged twice (2000  g) for 10 min. The dry sediment was mounted on a slide with
glycerine/gelatine slightly stained with an alcoholic solution of funchine. Slides were
microscopically observed and compared with the reference for identification. The following
terms were used for frequency classes: predominant pollen (more than 45% of pollen grains
counted), secondary pollen (16–45%), important minor pollen (3–15%), minor pollen (less than
3%). Thirty samples of each type of honey were analysed for moisture, specific gravity, total
acidity, optical rotation, ash and mineral contents. The samples were 6–8 months old and were
stored at 4
C in airtight glass containers till further analyses.

3. Physico-chemical analysis

The samples of honey were analysed according to the AOAC methods (AOAC, 1990) in order
to determine moisture, optical rotation, acidity (free, lactone and total) and ash content. Moisture
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in honey was determined with a refractometer reading at 20
C and obtaining corresponding %
moisture from the table (AOAC, 1990). Optical rotation was measured in a polarimeter as
follows: 10 g of honey sample was clarified with Carrez reagents (I and II) and distilled water was
added to get a final volume of 100 ml. Then, this solution was inserted into the polarimeter and
results were stated in angular on a 200 mm basis. Lactone and total acidity were determined by the
titrimetric method: first add 0.05 N NaOH and stop at pH 8.5 (free acidity), immediately pipette in
10 ml 0.05 N NaOH, and without delay back-titrate with 0.05 N HCL to pH 8.3 (lactone acidity).
Total acidity results from adding free plus lactone acidities (AOAC, 1990). Results were expressed
as meq/kg. The specific gravity of honey was determined by dividing the weight of specific gravity
bottle (50 ml) filled with honey by the weight of the same bottle, filled with water.

4. Mineral analysis

Ash content was measured by calcination, overnight at 550
C in a furnace, to constant mass
(AOAC, 1990). Sodium, potassium, calcium, iron, zinc and copper were determined using atomic
absorption spectrophotometer (Model AA670 Shimadzu, Kyoto, Japan) after wet digestion with
sulphuric acid and nitric acid following the method of Jacob (1958). In this method, honey sample
(5 g) was digested by heating in a Kjeldahl flask with concentrated nitric acid and sulphuric acid
mixture for oxidation of carbonaceous matter. For each sample, a blank was prepared
simultaneously by taking same amount of nitric and sulphuric acid mixture. Care was taken
during heating so that no excess foaming took place. Concentrated nitric acid in small amount
was added until all the organic matter was oxidized. This point was reached when no further
darkening of the solution occurred on continuous heating and a clear solution was obtained. It
was cooled and transferred to 100 ml volumetric flask and the volume was made up with distilled
water. The concentration of various metals was determined using atomic absorption spectro-
photometer (Model AA670 Shimadzu, Kyoto, Japan) by aspirating the solution into the oxygen–
acetylene flame. The instrument was calibrated by using standard solutions of various metal salts.

5. Statistical analysis

The data was assessed by analysis of variance (ANOVA) (Snedecor & Cochran, 1987) and by
Duncan’s multiple range test with probability Pp0:05 (Duncan, 1955).

6. Results and discussion

The pollen spectra of honey samples studied for physico-chemical and mineral content is briefly
described in the following section, and the percentages are relative to pollen of nectar-producing
plants. T. alexandrinum L honey contained 49–80% pollen of Trifolium sp. B. campestris
(mustard), honey contained 32–65% pollen of B. campestris. H. annuus (sun flower) honey
contained 16–72% pollen of H. annuus. E. lanceolatus honey contained 50–74% pollen of
Eucalyptus sp. Citrus honey contained 62–89% pollen of Citrus sp. Multi-flower honey contained
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Table 1
Statistical analysis of physico-chemical properties of different types of North Indian honey
Source Free acid Lactone Total acidity Moisture Specific Ash content Optical
(meq/kg) (meq/kg) (meq/kg) (%) gravity (% w/w) rotation


(g/cm2) a20 C
T. alexandrinum 21.9370.41a 17.6170.29a 39.5470.55a 18.6570.46a 1.3670.011* 0.1470.36a 9.3**
Citrus flower 32.6570.38b 14.7270.27b 47.3770.48b 18.4670.27b 1.3970.009* 0.1270.2b 8.4**
H. annuus 32.1470.47c 15.1870.42c 47.3271.97c 15.9270.44c 1.4270.012* 0.1770.23c 8.7**
E. lanceolatus 25.8470.31d 14.6470.30d 40.4870.51d 17.0970.45d 1.4270.008* 0.1670.3d 9.3**
B. campestris 14.5770.29e 15.4570.28e 30.0370.56e 14.6370.29e 1.4370.008* 0.2270.44e 7.5**
Multi-flower 16.6770.31f 14.7270.27f 31.3970.38f 13.9770.51f 1.4570.013* 0.2570.28f 8.2**
Commercial 19.7870.31g 18.6170.22g 38.3970.37g 17.8170.24g 1.4370.008* 0.2870.4g 7.2**
Values are means7s.d.; n ¼ 30:
Means not sharing a common superscript letter in column are significantly different at Pp0:05 as assessed by Duncan’s
multiple-range test.
*Non-significant at Pp0:01:
**Statistical analysis has not been performed.

2–5% pollen of B. rapa L. (toria); other pollen found were those from B. campestris var. toria,
sarson (B. campestris var. sarson) rayo (B. juncea) and Linum usitatissimum L. Commercial
sample contained 1–2% pollen of H. annuus, other pollens found were Brassica sp., Helianthus
sp., Eucalyptus sp., so it is also considered as multi-flower honey in the present investigation. The
results of physico-chemical characteristics of honey from different sources are summarized in
Table 1. Among the seven different sources statistically significant differences were found in
moisture content, free acid, lactone, total acidity (Po0:01) and non significant in specific gravity
(Po0:01). The moisture content of honey is an important factor contributing to its stability
against fermentation and granulation during storage. Trifolium honey showed the highest value
of moisture content as compared to other types. The mean values of moisture content of
Eucalyptus (17.09%) and Brassica (14.63%) honey were lower than the values 19.4%
(Eucalyptus) and 21.8% (Brassica) reported by Singh and Bath (1997). Estupinan and Sanjuan
(1998) reported the moisture content of honey in the range of 13–25% and stated that moisture
content was affected by climate, season and moisture content of original plant nectar. Multi-
flower honey showed the highest specific gravity and Trifolium honey the lowest. The honey
samples having a high moisture content had the lowest specific gravity and vice versa. Honey from
all sources showed the property of rotating the polarization plane of polarized light and optical
rotation was levorotatory in all samples of the seven types of honey (Table 1). Floral honey is
levorotatory in contrast to honeydew and some adulterated honey, which are normally
dextrorotatory. This is a consequence of normal preponderance of fructose in the floral honey,
which shows a negative specific rotation over glucose. The overall value for the optical rotation is
a result of the values of the different honey sugars. The measurement of specific rotation is
currently used in Greece, Italy and UK to distinguish between blossom and honeydew honeys.
Persano Oddo, Piazza, Sabatini, and Accorti (1995) found that blossom honey has negative values
of optical rotation, while honeydew honey has a positive one in Italian honey. Whether this
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Table 2
Statistical analysis of mineral content (mg/kg) of different types of North Indian honey
Source Ca Na K Zn Fe Cu
a a a a a
T. alexandrinum 84.6370.02 163.370.028 812.670.89 16.7770.028 10.2870.013 2.970.015a
Citrus flower 32.6070.014b 97.8770.015b 743.3770.64b 2.5570.018b 8.8670.015b 1.8870.016b
H. annuus 63.8170.017c 175.8370.014c 689.5770.23c 4.1670.013c 10.3970.016c 1.8970.018c
E. lanceolatus 67.3970.013d 177.670.020d 624.470.34d 4.6970.018d 12.5070.019d 2.2270.02d
B. campestris 56.7070.022e 304.370.031e 489.5270.19e 5.0570.024e 10.3570.020e 1.9570.017e
Multi-flower 72.9370.018f 247.1570.025f 932.5670.15f 11.2070.016f 10.1070.016f 1.8470.019f
Commercial 33.770.02g 194.970.040g 862.5270.19g 13.2570.020g 13.2570.020g 1.7470.023g
Values are mean7s.d.; n=30.
Means not sharing a common superscript letter in column are significantly different at Pp0.05 as assessed by Duncan’s
multiple-range test.

method is capable of differentiating honeys in other geographic regions of India remains to be

examined in future research.
Total acidity of Citrus and Helianthus honey was higher than the other honey types. Citrus
honey had the highest free acid content with the lowest value being for Brassica honey (Table 1).
The acidity of honey is due to the presence of organic acids, particularly the gluconic acid, in
equilibrium with their lactones or esters and inorganic ions such as phosphate and chloride
(Echingo & Takenaka, 1974). The variation in acidity among different honey types may be
attributed to variation in these constituents due to extraction season (Perez-Arquillue, Conchello,
Arino, Juan, & Herresa, 1994). El-Sherbiny and Rizk (1979) reported that total acidity was higher
in cotton honey than in clover honey which indicates the influence of floral types in total acidity.
The mineral content of honey from different sources along with the mean and corresponding
standard deviation are shown in Table 2. An analysis of variance for the mineral content results
showed that the average values are significantly different (Po0:01). One might explain the
significant differences in mineral content as owing to the different botanical origins of honey.
However, other factors such as geographical conditions are also expected to affect the mineral
content. The similar results in other types of honey at different locations were observed by several
workers (Salinas, Montero De Espinosa, Osorio, & Lozano, 1994; Frias, Hardisson, Gonzalez,
Munoz, & Espinar, 1997; Russo-Almeida, 1997). The North Indian honey had lower ash content
than that reported from South Spain (Serra & Ventura, 1995) commercial Spanish honey
(Rodriguez-Otero et al., 1992) and Galicia (North-West Spain) (Rodriguez-Otero et al., 1994).
The range of values for ash content (0.12–0.28%) fell within the limit allowed for floral honey
(0.6%) and indicated the cleanness of honey samples and possibly the lack of adulteration with
molasses (Krauze & Zalewski, 1991). Potassium was the most abundant of the elements present in
all honey types while copper was present in the lowest amount. Multi-flower honey was the best
source of potassium and sodium with mean values of 932.56 and 247.15 mg/kg, respectively. Mean
values of potassium content varied between 932.15–489.52 mg/kg. Rodriguez-Otero et al. (1994)
reported potassium as the most abundant element, with an average content of 1500 mg/kg with
mean content of sodium, calcium, copper, and iron as 138, 102, 1.11 and 5.12 mg/kg, respectively,
in Galicia (North-West Spain) honey. Commercial honey was the best source of iron followed by
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Eucalyptus and Helianthus. A mean value of iron content ranged between 8.86 and 13.25 mg/kg.
Trifolium was the best source of calcium and zinc with an average content of 84.63 and
16.77 mg/kg, respectively. Citrus honey had the lowest value for calcium and zinc.

7. Conclusion

It could be concluded from the above results that moisture content, free acid, lactone, and total
acidity are the physico-chemical properties that depend upon the flower types used by honeybees
for nectar. The measurement of specific rotation can be used to distinguish between blossom and
honeydew honeys produced in Northern India. Potassium is the most abundant of the elements,
while copper is the least present element in the Northern Indian honey. Mineral content of honey
is highly dependent on the type of flower used by bees for nectar. Extensive research is required
to establish physico-chemical properties and mineral content variations according to the
geographical area.

Acknowledgements

The first author acknowledges the kind help and facilities provided by the technical staff of
Cereal Technology Laboratory of the Defense Food Research Laboratory, Mysore (India).

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