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Journal of Integrative Medicine ›› 2014, Vol. 12 ›› Issue (4): 379-389.doi: 10.1016/S2095-4964(14)60033-9

• Research Article • Previous Articles     Next Articles

Investigation of the nutraceutical potential of monofloral Indian mustard bee pollen

Sameer S. Ketkara, Atul S. Rathorea, Sathiyanarayanan Lohidasana, Lakshmi Raob, Anant R. Paradkarc, Kakasaheb R. Mahadik a   

  1. Centre for Advanced Research in Pharmaceutical Sciences, Poona College of Pharmacy, Bharati Vidyapeeth University, Pune-411038, India
    Central Bee Research and Training Institute, Pune-411016, India
    c Centre for Pharmaceutical Engineering Sciences, University of Bradford, Bradford, West Yorkshire, BD7 1DP, UK
  • Received:2014-03-14 Accepted:2014-05-14 Online:2014-07-10 Published:2014-07-15


This study was designed to investigate the nutraceutical potential of monofloral Indian mustard bee pollen (MIMBP). 


The nutritional value of MIMBP was examined in terms of proteins, fats, carbohydrates, and energy value. Its chemical composition in terms of total polyphenol and flavonoid content was determined. MIMBP was screened for free flavonoid aglycones by developing and validating a high-performance liquid chromatography-photo diode array (HPLC-PDA) method. MIMBP was analyzed for in vitro antioxidant effect in terms of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging activity. 


MIMBP was found to be comprised of proteins ((182.2±5.9) g/kg), fats ((137.7±6.8) g/kg) and carbohydrates ((560.6±17.4) g/kg), which result in its high energy value ((17 616.7±78.6) kJ/kg). MIMBP was found to contain polyphenols ((18 286.1±374.0) mg gallic acid equivalent/kg) and flavonoids ((1 223.5±53.1) mg quercetin equivalent/kg). The HPLC-PDA analysis revealed the presence of kaempferol ((65.4±0.5) mg/kg) and quercetin ((51.4±0.4) mg/kg) in MIMBP, which can be used as markers for determining the quality of bee pollen. The MIMBP extract showed DPPH free radical-scavenging activity with a half maximal inhibitory concentration of 54.79 μg/mL. 


The MIMBP was found to be a rich source of nutrients providing high caloric value, which makes it a candidate for a potential nutraceutical agent. The study also illustrated the high antioxidant content of MIMBP, especially in the principle polyphenols and flavonoids, which suggests its potential role in the prevention of free radical-implicated diseases. The DPPH-scavenging effect of MIMBP further confirmed its antioxidant potential. Additionally, we developed a simple, specific and accurate HPLC-PDA method for the identification and quantification of free flavonoid aglycones. This can be applied in future screenings of the quality of pollen collected by honeybees.

Key words: Dietary supplements, Bee pollen, Flavonoids, Mustard plants

Figure 1

Light microscopic images of monofloral Indian mustard bee pollen A: Untreated pollen without acetolysis (40×); B: Acetolyzed pollen showing reticulate ornamentation (40×); C: Scanning electron microscope image of monofloral Indian mustard bee pollen (154×)"

Figure 2

HPLC-PDA chromatogram of (A) standards, (B) flavonoid aglycone compounds from the MIMBP sample solution HPLC-PDA: high-performance liquid chromatography-photo diode array; MIMBP: monofloral Indian mustard bee pollen."

Figure 3

UV spectra for standards (A) rutin, (B) chrysin, (C) kaempferol, and (D) quercetinThe HPLC method for quantification of flavonoid aglycones was validated and showed good linearity (r2 > 0.998) in the concentration range of 10-100 μg/mL, which was wide enough to quantify constituents in the MIMBP sample solution. Results for linearity of calibration curves, LOD, LOQ, precision, repeatability, specificity, and accuracy along with ANOVA and residual analysis are summarized in Table 1. RSD values for all standards in the range of 0.22 to 1.76 indicated that the method exhibited acceptable intraday and interday variation with respect to working standards. The accuracy as measured by the recovery % with small %RSD ranged from 98.52% to 100.06%. No peak interference at the retention times for standards and sample solution indicated specificity of method. The peak purity factors generated using PDA detector for aglycone peaks were within threshold values indicating no additional co-eluting peaks in the standard and sample solutions."


Figure 4

Overlay spectra of peaks of (A) standard kaempferol and sample solution at rentention time 12.097 min, and (B) standard quercetin and sample solution at rentention time 10.519 min"

Figure 5

DPPH-scavenging effect of MIMBP extract Data are expressed as mean ± standard deviation, n=3; *P<0.05, vs MIMBP extract solution. MIMBP: monoflora Indian mustard bee pollen; DPPH: 2,2-diphenyl-1-picrylhydrazyl."


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