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							| Functional Foods in Health and Disease - Online ISSN: 2160-3855; Print ISSN: 2378-7007



	

				
						
 
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																Functional Foods in Health and Disease - Online ISSN: 2160-3855; Print ISSN: 2378-7007
										

				


				
				
					
					
						
																					
				
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					Vol. 5 No. 6 (2015): June 2015
				
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									Research Articles
							
		



	
		  	 

		
	
		A water-soluble high molecular weight substance isolated from Hyuganatsu orange (Citrus tamurana), suspected to be a polysaccharide,  inhibits rat osteoclast cell formation
	


	
	
		

							
					Authors

						
							
								Hiroko Hata
							
															
									Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
																	
																																		
	
							
								Masatoshi Yamaguchi
							
															
									Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
																	
																																		
	
							
								Hiroshi Sameshima
							
															
									Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
																	
																																		
	
							
								Tsuyomu Ikenoue
							
															
									Department of Obstetrics and Gynecology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake-Cho, Miyazaki City, Miyazaki 889-1692, Japan
																	
																																		
	
							
								Junko Matsubara
							
															
									Ichimaru Phalcos Company, 
318-1, Asagi, Motosu City, Gifu 501-0475, Japan
																	
																																		
	
							
								Makoto Tsuboi
							
															
									Ichimaru Phalcos Company, 
318-1, Asagi, Motosu City, Gifu 501-0475, Japan
																	
																																		
	
							
								Takashi Tanaka
							
															
									Laboratory of Natural Product Chemistry, 
Nagasaki University Graduate School of Biomedical Sciences, 1-14 Bunkyo-cho, Nagasaki City, Nagasaki 852-8521, Japan
																	
																																		


				
			
																	
					
												DOI:
					

					
						
							https://doi.org/10.31989/ffhd.v5i6.182
						
					
				
			

						
										
					Abstract

					Background: Osteoporosis is detrimental to aged women’s health care. We previously reported that Hyuganatsu orange (Citrus Tamurana) contains active substances that inhibit osteoclast activities. Prior to conducting a human study, we sought to identify the biological active substance in the Hyuganatsu orange which suppresses osteoclast formation.
Methods: We isolated five fractions from a Hyuganatsu orange extract according to molecular weight. Each fraction was tested to determine its suppressive effect on the formation of osteoclasts in rats. We also used high-performance liquid chromatography (HPLC), infra-red (IR), and 1H and 13C NMR spectroscopy to evaluate its chemical structure. Data was recorded as mean ± standard error of the mean. The Mann-Whitney test was used, and a p-value of <.05 was considered statistically significant.
Results: The highest and lowest molecular weight fractions showed significant suppression activity on rat osteoclast formation (p < .05). The lowest molecular weight fraction was identified as hesperidin using thin layer chromatography. Additionally, IR absorption revealed that the highest molecular weight fraction was not a flavonoid. With regard to chemical structure, 1H and 13C NMR spectroscopy suggested that the highest molecular weight fraction had signals compatible with a polysaccharide such as galacturonic acid.
Conclusions: Hyuganatsu orange contains a biological active substance other than hesperidin that may be a polysaccharide and may suppress osteoclast formation.
Key Words: Citrus Tamurana, TRAP positive cell, Polysaccharides, Rat osteoclast cell, in vitro study

				
			
			

						
																																																													
						
		


		

						
										
					
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																			2015-06-01
																	

				
							

			
						
										

											
							
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									Vol. 5 No. 6 (2015): June 2015
								
							

						
					
											
							
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								Research Articles
							

						
					
									

			
						
										
					
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