Preliminary characterization of two important Honeys produced by Apis cerana and Apis dorsata from five Districts of Manipur, India

377 Preliminary characterization of two important Honeys produced by Apis cerana and Apis dorsata from five Districts of Manipur, India Elangbam Monika Devi, P. K. Singh and Thangjam Anand Singh * Department of Biochemistry, Manipur International University (MIU), Airport Road, Ghari Awang Leikai, Imphal West, Manipur, India). World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 377-383 Publication history: Received on 03 October 2025; revised on 10 November 2025; accepted on 13 November 2025 Article DOI: https://doi.org/10.30574/wjbphs.2025.24.2.1006 Abstract Honey, the natural sweet substance produced by honeybees is a valuable product with significant nutritional and medicinal importance. Our study evaluates the physicochemical parameters of honey produced by Apis cerana and Apis dorsata from five districts of Manipur: Imphal East, Imphal West, Thoubal, Bishnupur, and Kakching. The parameters included moisture content, pH, and specific gravity, following standard laboratory methods. The moisture content ranged between 13.9% and 20.9% for A. cerana and 15.5%-17.5% for A. dorsata. The pH values varied from 3.30-4.07 and 3.59-4.05, respectively, while specific gravity ranged from 1.25-1.41 and 1.34-1.40. These results show that, all samples are in line the Codex Alimentarius standards for high-quality honey, with small variations linked to floral and geographical influences. These findings establish a foundation for assessing the quality of regional honeys from Manipur. Keywords: Honey; Apis cerana; Apis dorsata; Physicochemical properties; Manipur 1. Introduction Honey is only a natural sweetener but also have functional food valued for its biological activity and nutritional properties. Its composition is largely influenced by many factors such as bee species, different types of flowers, place of origin, harvesting method, and post-harvest handling practices [1,2,3]. By its rich nutritional and therapeutic properties, honey has been widely utilized in traditional and modern medicine for its antioxidant, antimicrobial, anti-inflammatory, and wound healing activities [4,5]. These properties are strongly correlated with the physicochemical characteristics of honey, which are wide used as quality determinants for commercial rating and authorization [6]. The physicochemical parameters of honey include colour, moisture content, pH, electrical conductivity, specific gravity, ash content, total soluble solids, HMF, proline, free acidity, sugar composition [7]. Among these, moisture content plays an important role in indicating honeys longevity, if the moisture content is high, increase the risk of fermentation by high resistant of yeasts [8]. In the same way, pH determines honey flavour, stability, and antimicrobial activity, with acidic pH contributing to microbial spoilage [9]. Specific gravity is another important parameter, as it depicts honey density and concentration, as a result indicating its purity and maturity [10]. These parameters are commonly analysed according with international standards established by the Codex Alimentarius Commission and European Union legislation, which set standard limits for high quality honey [11]. In India, honey is majorly produced by native bee species such as Apis cerana and wild species Apis dorsata, both of which contribute to distinctive honey types with varying physicochemical characteristics due to differences in foraging behaviour and hive environment [12]. A. cerana is a domesticated species while A. dorsata, also called as the giant honeybee, generally builds their nests in forest areas as well rural and urban areas, giving to variations in nectar source World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 377-383 378 and resulting honey quality [13]. Several studies have reported that according to different species of bee and geographical region different type of honey is notable with variation in pH, specific gravity and moisture’s, it indicates the importance of regional characterisation and authenticity of honey quality [14-16]. Manipur is the Northeastern part of India, is enriched with floral richness and favourable climatic conditions suitable for honey production from both wild bees as well domesticated bee species. However, in Manipur, the scientific comparative assessment data regarding honey produced by A.cerana and A.dorsata from the five district. So, such kind of information is required for quality control, commercial standardization, and promoting the regional branding of Manipur honey in national and international markets. Therefore, this study aims to comparatively analyse the moisture content, pH, and specific gravity of honey produced by Apis cerana and Apis dorsata collected from five districts of Manipur: Imphal East, Imphal West, Thoubal, Bishnupur, and Kakching. So, this scientific finding is hoped for the understanding the corelation with the bee species, geographical origin and the honey quality to contribute the developing the regional honey. 2. Materials and Methods 2.1. Study Area The study was conducted in five valley districts of Manipur, India; Imphal East, Imphal West, Thoubal, Bishnupur, and Kakching. It is located at the heart of Manipur. These regions are surrounded by hills with a moderate subtropical monsoon climate. Due to this diversity of plants, richness of flora supporting both domesticated and wild honeybee populations [17]. It lie 790-830 metres above the sea level and by this bring the ecological variations within these districts and contribute to differences in nectar sources, which may influence honey composition [18]. Figure 1 Map of Manipur Figure 1a Five valley Districts of Manipur 2.2. Sample Collection The 5 (five) honey samples were collected from colonies of Apis cerana (domesticated) maintained in traditional and modern wooden hives, and 5 (five) from wild colonies of Apis dorsata. Total 10 samples collections were performed in between December 2023 to February 2025 under aseptic conditions using sterilized containers. For each district, freshly harvested honey was sourced directly from local beekeepers (A. cerana) and wild honey hunters (A. dorsata). All samples were stored in airtight glass jar and kept at the laboratory in room temperature for analysis, assuring without exposure to heat and minimum light to prevent alteration of physicochemical properties [19]. 2.3. Materials requirements Abbe-type digital refractometer, Thermometer, Digital Phmeter, Analytical balance, Lint free wipe, specific gravity bottle. World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 377-383 379 2.4. Physico-chemical Analyses 2.4.1. Moisture Content Moisture content was determined using an Abbe refractometer (NAR-2T, Atago Co., Ltd., Tokyo, Japan) [22], following the method recommended by the International Honey Commission (IHC). With the help of refractometric conversion tables, interpreted the moisture percentage of honey by setting the refractometer at 20˚C [20]. Table 1: Moisture percentage of honey of A.cerana and A. dorsata District Apis cerana (%) Apis dorsata (%) Imphal East 18.6 15.5 Imphal West 13.9 15.9 Thoubal 20.9 15.9 Bishnupur 16.3 17.5 Kakching 18.2 17.1 2.4.2. pH Determination The pH of honey samples was measured using a calibrated digital pH meter (Hanna Instruments, USA). For each measurement, 10 g of honey was dissolved in 90 mL of distilled water and stirred gently before measurement, following AOAC guidelines [21]. Table 2: pH content of honey of A. cerana and A. dorsata District Apis cerana Apis dorsata Imphal East 3.91 3.59 Imphal west 4.07 4.05 Thoubal 3.30 3.98 Bishnupur 3.74 3.87 Kakching 3.49 3.77 2.4.3. Specific Gravity Specific gravity was determined using a specific gravity bottle (pycnometer) method. The empty bottle was weighed, then filled with distilled water as a reference, and subsequently with each honey sample. Specific gravity was calculated using the formula SG = (Weight of honey filled bottle - Weight of empty bottle) / (Weight of waterfilled bottle - Weight of empty bottle), in accordance with Codex Alimentarius standards [22]. Table 3: Specific gravity value of A.cerana and A.dorsata District Apis cerana Apis dorsata Imphal East 1.369 1.397 Imphal West 1.412 1.399 Thoubal 1.250 1.400 Bishnupur 1.400 1.342 Kakching 1.377 1.360 World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 377-383 380 2.5. Statistical Analysis The data comparison between A. cerena and A. dorsata were analysed by ANOVA (Analysis of variance). Statistical analysis was performed using SPSS version 25.0 (IBM Corp., USA). The statistical significance and results were expressed in mean values. It was considered at p < 0.05 [23]. 3. Results The significant variations of physicochemical characteristics including moisture content, pH and specific gravity of A. cerana and A. dorsata from five Districts of Manipur is due to the differ in bee species and geographical origin. 3.1. Moisture Content The moisture content of A. cerana honey samples ranged between 13.9% and 20.9%, while A. dorsata honey showed a narrower range of 15.5% to 17.5%. The highest moisture content in A. cerana was observed in samples of Thoubal District collected from humid regions, suggesting greater environmental influence on domesticated bee honey. Comparatively, A. dorsata honey exhibited relatively stable moisture levels, which may be attributed to its wild nesting behaviour and advanced natural dehydration prior to harvesting. ANOVA results indicated a statistically significant difference (p < 0.05) in moisture content between the two bee species and across districts. 3.2 pH Values The pH values of A. cerana samples ranged from 3.30 to 4.07, while A. dorsata honey exhibited pH values between 3.59 and 4.05. Overall, A. dorsata honey have a slightly higher mean pH compared to A. cerana honey. District wise variations were observed in both species, reflecting influence from floral sources. The acidic nature of all samples is reliable with reported values for high quality honey, suggesting good microbial stability. Statistical analysis confirmed significant (p < 0.05) species and region wise differences in pH. 3.2. Specific Gravity The specific gravity of A. cerana honey ranged from 1.25 to 1.41, whereas A. dorsata samples showed specific gravity values between 1.34 and 1.40. The slightly higher specific gravity in A. dorsata honey samples indicates a denser composition, potentially due to increased sugar content and lower moisture variation. District level variations suggested that honey collected from areas with abundant floral resources had comparatively higher density. Significant differences (p < 0.05) were recorded between species and among districts. 3.3. Overall Trends When compared with honey from Apis dorsata indicate more stable physicochemical values across all parameters, while Apis cerana honey showed wider variation, likely due to differences in hive management and nectar source stability. All parameters measured ranged between the acceptable international standards for honey quality, indicating that both honey types from Manipur fulfil global quality requirements. 4. Discussion The present study indicated that moisture content in Apis cerana honey (13.9%-20.9%) was more variable compared to Apis dorsata honey (15.5%-17.5%). These findings are closely resembled with the Nepal and India. The moisture ranges of Nepal honey in A. cerana are varied between 14.2% and 21.0% [24,25]. The comparatively narrower range observed in A. dorsata honey correspond with studies conducted in Thailand and Indonesia, where giant bee honey exhibited moisture content between 15.0% and 18.0% due to its natural dehydration process in exposed combs [26,27]. Higher moisture in some A. cerana samples may be attributed to premature harvesting and climatic humidity variations, as reported in regional studies from north eastern India [28]. This indicates that harvesting practices and hive management significantly influence moisture variability in domesticated bee honey. The pH values observed in the current study (3.30-4.07 for A. cerana and 3.59-4.05 for A. dorsata) are consistent with the acidic nature of honey reported globally, which typically ranges between 3.2 and 4.5 [29]. The slightly higher pH in A. dorsata honey corresponds with findings from studies conducted in Malaysia and Bangladesh, where wild bee honeys show higher acidity stability due to nectar source diversity and enzyme content [30,31]. Lower pH in some A. cerana samples supports earlier findings that domesticated bee honey may exhibit greater acidity fluctuations due to World Journal of Biology Pharmacy and Health Sciences, 2025, 24(02), 377-383 381 environmental and floral source influences [32]. The acidic pH range in all samples indicates strong antimicrobial potential and increased shelf life, consistent with international quality standards [33]. Specific gravity ranged between 1.25 and 1.41 for A. cerana and 1.34 and 1.40 for A. dorsata, indicating denser composition in wild bee honey. These values are similar to those reported in African A. dorsata honey studies, with specific gravities from 1.34 to 1.41 [34], and Indian A. cerana honey ranging between 1.28 and 1.39 [34,35]. The higher density in A. dorsata samples may be associated with higher sugar concentration and reduced moisture variability, as observed in earlier comparative honey studies [36]. Additionally, geographical influence was evident, with honey from florally diverse districts exhibiting higher specific gravity, supporting findings from regional analyses that floral composition affects honey density and solute concentration [37,38]. The data differences between A. cerana and A. dorsata honey support previous literature suggesting that wild bee species, due to their larger body size and foraging behaviour, likely to produce honey with higher sugar concentration and lower moisture fluctuations than domesticated species [38,42]. Geographical and floral variations across Manipur districts also likely contributed to the variation of data, consistent with reports indicating that climatic conditions, soil nutrient content, and floral diversity significantly affect honeys physicochemical properties [39,40,41,43]. The consistency of all parameters with Codex Alimentarius and EU standards confirms that both honey types from Manipur possess high commercial quality and export potential [11]. As a whole, the finding gives the importance of species and specific interaction and the influence of geographical region, hive management, foraging behaviour in honey. So, reason-based profiling help to establishing of producing quality honey in Manipur and the consumers also trust the regional honey. 5. Conclusion The present comparative study of honey produced by Apis cerana and Apis dorsata from five districts of Manipur: Imphal East, Imphal West, Thoubal, Bishnupur, and Kakching show the significant differences in their physicochemical characteristics. Moisture, pH, and specific gravity values fell within international quality standards, confirming that both species produce honey of acceptable commercial and nutritional quality. However, A. dorsata honey showed relatively stable moisture content, higher specific gravity, and slightly higher pH, suggesting superior stability and lower fermentation risk compared to A. cerana honey. Geographical variations also influenced the parameters, demonstrating that local floral composition and climatic conditions play a major role in honey composition. These results emphasize the potential for regional honey classification and the development of quality control protocols for Manipur honey. 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