15 Nippon Shokuhin Kagaku Kogaku Kaishi Vol /0 No 3 2 23 (**3) 483 + + + * * * * * * * * * + * * * * Inhibition of Fat Digestion by Highly Polymeric Proanthocyanidins from Seed Shells of Japanese Horse Chestnut ( B LUME) Aesculus turbinata + + + Hideto Kimura* Satoshi Ogawa* * Ai Niimi* Mitsuo Jisaka* * Takuya Katsube* and Kazushige Yokota* * + * Department of Research and Development Kotobuki Seika Co Ltd *2 Hatagasaki Yonago Tottori 02 *2/ * The United Graduate School of Agricultural Sciences Tottori University +*+ Koyamaminami Tottorishi Tottori 02* 2// * Department of Life Science and Biotechnology Faculty of Life and Environmental Science Shimane University +*0* Nishikawatsucho Matsue Shimane 03* 2 * Shimane Institute for Industrial Technology + Hokuryocho Matsue Shimane 03* *2+0 Seed shells are discarded as waste products of Japanese horse chestnut seeds used as ingredients in traditional Japanese foods Hotwater extraction of these seed shells yielded substances that were tested in vitro for their inhibitory e# ects on pancreatic lipase The extracts were fractionated by column chromatography on Diaion HP* and Chromatorex ODS +* T columns to obtain the polyphenolic compounds The polyphenolic compounds were separated by Sephadex LH* column chromatography into three fractions (F + F and F ) depending on their degree of polymerization F + containing the phenolic substances with the lowest molecular weight failed to inhibit pancreatic lipase whereas F and F both showed potent inhibitory activities to pancreatic lipase An oral fat tolerance test in mice revealed that these polymeric proanthocyanidins suppressed fat digestion in vivo Taken together seed shells of Japanese horse chestnuts may provide a source for highly polymerized polyphenols that could be used as nutraceutical factors with antiobesity e# ects (Received Apr + **3 ; Accepted Jun +/ **3) Keywords : high polymeric proanthocyanidin Japanese horse chestnut Aesculus turbinata B LUME polyphenolic compound fat digestion pancreatic lipase : Aesculus turbinata BLUME Aesculus turbinata BLUME Aesculus hippocustanum L + * 02 *2/ *2 * 02* 2// +*+ * 03* 2 +*0* * 03* *2+0 + Corresponding author hkimura@kozuchinetjp +
484 /0 3 **3 3 16 +* T * ml * ml 1* * ml 1 FolinCiocalteu /0 F F GPC / 23 /0 +* g +** mg Sephadex LH* +** mm +* mm GE Healthcare UK Ltd Buckinhamshire UK ml F+ F 1* F ao Shodex GPC ao STANDARD SL+*/ Mw 0 /2* +++* + +1* *1* * /+* 1+* +*** +30** F Mw 1** in vitro in vivo B Mw /12/ A Mw /10/ FolinCioalteu 33 / :*/ vv * 0 mlmin methylumbelliferyl oleate B * 2* nm A type Sigma St Louis MO USA GPC type **00 Ao GPC F F Diaion HP* 0 HPLC LC*+* A CR 2A TSKgel Super + AW *** + mm 0 * mm Aes N N M culus turbinata B LUME / Shodex GPC M methylumbelliferyl oleate STANDARD SL+*/ Mw /2* +30** 1 * ** F Mw 1** E C * mm Mcllvaine ph 1 ** l *+ M + ml *+ M ml FP 111 JASCO * nm nm methylum belliferone +** +* Han 2 ODS Chromatorex ODS ICR
17 : 485 0 ICR MF + 0ml 2* mg mg 0ml +0 * Diaion HP* Chromatorex ODS +* T 2 +** l + kg * +** ** * mg +** l 1/ * *** g E + + Sephadex LH* F + fraction number * * ; F fraction number + * ; F fraction number + 0* 0 IC + gml * mgkg + ** gml IC Dunnett P**/ 1 1 3 Diaion HP* +* T / ICR Sephadex LH* + F+ F F + F 0 F + F F MF +0 Chromatorex ODS /0 0 B A + kg *** mg 0 + 1 +* + + MALDITOFMS F +3 mer F mer MALDITOFMS /0 GOT GPT C F F GPC GPC + F F F + *** F + *** F F / +*2
486 /0 3 **3 3 18 GPC F F F F A B F F GPC GPC A Mw /10/ B Mw /12/ /2* F F IC *2 gml *3 gml F F F+ F F F + ; F ; F n
19 : 487 / / gml B 0 3 A A IC B A Nakai +* / 0 ; ; ; ; B ; A n 0 2 *1 gml IC 3 ++ Sugiyama HPLC *3 gml IC 2 IC +/ gml 3 gml 0 1 gml / + gml A / 0
488 /0 3 **3 3 20 1 2 : ; +** mgkg ; ** mgkg : *** mgkg ; * mgkg : n0 : *** mgkg n0 * P**/ Diaion HP* Chromatorex ODS +* T 2 + kg ** mg / * mg + / / 1 Diaion HP* + + GOT GPT *** mgkg Karmen unit GOT 141 4+ /243 40* GPT *4/ 4+ 240 +4+ GOT /14040 /341 4 GPT 34+ +4 +43 +4 + **1 ICR Diaion HP* Cromatorex ODS +* T n0 * P*4*/ Chromatorex ODS +* T *** mgkg 2 + Diaion HP* GOT GPT Chromatorex ODS +* T Sepha + GOT dex LH* F+ F F GOT / 0 2 +* 1+ +/ 0 + *** F + *** F F F GOT GPT F F B A + kg ** mg * mg
21 : 489 / Ogawa S Kimura H Niimi A Jisaka M Katsube T and Yokota K Fractionation and structural characterization of polyphenolic antioxidants from seed shells of *** mgkg Japanese horse chestnut ( Aesculus turbinata B LUME) J GOT GPT Agric Food Chem /0 +*0 +*/+ (**2) 0 /0 3/ +* **3 1 JulkunenTiitto R Phenolic constituents in the leaves of northern willows : methods for the analysis of certain phenolics J Agric Food Chem / + +1 ( +32/ ) 2 Han LK Kimura Y Kawashima M Takaku T Taniyama T Hayashi T Zheng YN and Okuda H + Yoshikawa M Murakami T Matsuda H Yamahara Antiobesity e# ects in rodents of dietary teasaponin a J Murakami N and Kitagawa I Bioactive saponins lipase inhibitor Int J Obes Relat Metab Disord / and glycosides III horse chestnut ( + ) : The structures +/3 +0 (**+) inhibitory e# ects on ethanol absorption and hypogly 3 cemic activity of Escins Ia IIa Ib IIb and IIIa from the // 132 seeds of Aesculus hippocastanum L Chem Pharm Bull **2 +/ +0 ( +330 ) +* Nakai M Fukui Y Asami S ToyodaOno Y Iwashita T Shibata H Mitsunaga T Hashimoto F and Kiso Y Inhibitory e# ects oolong tea polyphenols on pancreatic /+ 01 013 ** lipase in vitro J Agric Food Chem / /3/32 (**/) ++ Sugiyama H Akazome Y Shoji T Yamaguchi A Yasue M Kanda T and Ohtake Y Oligomeric procyanidins / + 2 in apple polyphenol are main active compo **0 nents for inhibition of pancreatic lipase and triglyceride Kimura H Ogawa S Jisaka M Kimura Y Katsube absorption J Agric Food Chem // 0* 0*3 (**1) T and Yokota K Anitobese e# ect of novel saponins + **1 http : // from edible seeds of Japanese horse chestnuts ( Aesculus jslccojp/inform/ **1 data/icr **1pdf turbinata B LUME) after treatment with wood ashes J Agric Food Chem /0 12 122 (**2) + + + 0 +/