Journal of Integrative Agriculture 2016, 15(0): Available online at ScienceDirect

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1 Journl of Integrtive Agriulture 2016, 15(0): Aville online t SieneDiret RESEARCH ARTICLE Quntifition nd nlysis of nthoynin nd flvonoids ompositions, nd ntioxidnt tivities in onions with three different olors ZHANG Shi-lin, DENG Peng, XU Yu-ho, Lü Shn-wu, WANG Jin-jun Stte Key Lortory of Crop Genetis nd Germplsm Enhnement, Key Lortory of Biology nd Germplsm Enhnement of Hortiulturl Crops in Est Chin, Ministry of Agriulture/College of Hortiulture, Nnjing Agriulturl University, Nnjing , P. R.Chin Astrt Onion (Allium ep L.) is very importnt vegetle rop widely ultivted throughout the world. In this study, we mesured totl nthoynins nd flvonols of onions of three olors. The fresh weight of totl nthoynins were (29.99±1.19), (9.64±1.30) nd (0.75±0.40) mg 100 g 1 fruit in red, yellow nd white onions, respetively. Likewise, the fresh weight of totl flvonoids were (111.10±5.98), (36.64±3.59), nd 0 mg 100 g 1 in red, yellow nd white, respetively. Four types of nthoynins (delphinidin 3,5-diglyosides, ynidin 3,5-diglyosides, ynidin 3-glyosides nd ynidin 3-(6 -mlonyl)-gluopyrnoside) nd two kinds of flvonoids (queretin nd queretin 3-glyosides ) were identified in two vrieties (red nd yellow ul) of onions y HPLC/DAD ESI/MS. The totl polyphenol ontents were lso mesured y mens of Folin-Ciolteu method. Moreover, ll the onentrtions of nthoynins, flvonoids nd polyphenols showed signifintly positive orreltions with ntioxidnt tivities mesured y DPPH, ABTS + nd FRAP ssys. This study provided informtion on nthoynin nd flvonoids ompositions tht will e useful for onion reeding. Keywords: onion (Allium ep L.), nthoynins, flvonoids, ntioxidnt tivity, HPLC-DAD ESI/MS 1. Introdution Onion (Allium ep L.) (2n=16), one of very importnt griulturl eonomi rops, is widely plnted round the glol s vegetles with round 640 million tons of outputs tht is only surpssed y tomto nd wtermelon (Brewster Reeived 20 Novemer, 2015 Aepted 26 April, 2016 ZHANG Shi-lin, Moile: , E-mil: @ qq.om; Correspondene WANG Jin-jun, Tel: E-mil: wngjinjun@nju.edu.n 2016, CAAS. All rights reserved. Pulished y Elsevier Ltd. doi: /S (16) ; Mllor 2014). Nowdys, ul olor hs eome prominent trit in onions s people give top priority to the helthy funtion of vegetles, due to the undnt flvonoid ontent nd the enefiil effets of nutritious ntioxidnts presene in them (MDougll 2007; Sunggil 2009). There re three mjor olors of onion germplsm, white, yellow nd red, whih re due primrily to nthoynins in the epiderml ells of the sle leves of the ul (Donner 1997). Anthoynins, produing lue, red or purple olors, group of nturl plnt pigments tht re widely distriuted in vegetles (Mrkkis 1982), inluding onion, red ge, Chinese red ge, lettue nd eggplnt (Bridle 1997; Clifford 2000). Anthoynins nd flvonoids with espeil hemil struture led to retion towrds the free rdils power, nturl ntioxidnts, ntiinflmmtory,

2 ZHANG Shi-lin et l. Journl of Integrtive Agriulture 2016, 15(0): ntiner properties nd Alzheimer s nd Prkinson s diseses (Wng nd Mzz 2002; Hou 2003; Kong et l. 2003; Glvno 2005; Yin et l. 2015). People inresingly intend to hoose vegetles whih re rih in nthoynins. By fr, the reserh of nthoynins in onions hs quired mny signifint hievements nd out 10 kinds of nthoynins hve previously een identified inluding four min nthoynins (3-(3 -gluosyl-6-mlonylgluoside), 3-(6 -mlonylgluoside), the 3-(3 -gluosy- lgluoside) nd 3-gluoside of ynidin ) nd nother six minor nthoynin pigments (Fossen 1996; Donner 1997). Flvonoids re lrge nd diverse group of polyphenoli ompounds with ntioxidnt effets (Pérez 2014). There were severl investigtions into flvonoids in onions, suh s queretin-3-o-rutinoside, queretin 4-O-β-gluoside nd queretin 3,4-O-β-digluoside (Hollmn nd Arts 2000; Murot nd Tero 2003), in whih ount for out 80% of the totl flvonoid ontents(bonorsi et l. 2005). Onion is widely ultivted in mny regions of Jingsu Provine in Chin s one of the min vegetles. In this study, we hose 628, W470 nd W1201 onion ultivrs with white, yellow nd red, respetively, whih were most populr in middle-est of Chin s the sme tegory of dy neutrl onions. HPLC-ESI-MS n seprte nd identify nthoynins nd flvonoids with high effiieny nd high qulity. Wht s more, DPPH (1,1-diphenyl-2-pirylhydrzyl free rdil), ABTS + (2,2 -zino-is (3-ethyl-enzothizoline-6-sulfoni id) dimmonium slt), FRAP (ferri reduing ility of plsm) ssys, in whih re widely nd ommonly used methods involving the determintion of the dispperne of free rdils, were used to evlute the ntioxidnt tivities of onion with three different olor smples in our study. There hve een severl reports out the nthoynins in red onion, however, the nthoynin omposition nd ontent of white nd yellow onion hve still not een fully hrterized nd the ntioxidnt tivity of onions with three different olor is short of omprehensive omprison. Therefore, we here provide detiled ssessment of the ontents nd ntioxidnt tivities of nthoynin mong three different olor onions. 2. Mterils nd methods 2.1. Chemils nd mterils Three different olor onions 628 (white), W470 (yellow) nd W1201 (red) were hrvested in the Jinpu Frm (32 00 N, E), Nnjing, Jingsu Provine, Chin, in My 2015, pled in polyethylene gs nd stored t 70 C until hromtogrphi nlysis. The plnt ws lssified t the College of Life Siene, Nnjing Agriulturl University, Chin (Fig. 1). All solvents were of nlytil grde Extrtion of nthoynins nd flvonoids Extrtion of nthoynins ws performed ording to the method desried previously y Long (2007) with some modifitions. 10 g fresh smples were treted with homogenizer in 4 C, nd then ellulr struture ws roken using high-speed homogente mhine t r min 1, 2 min. Then we poured the onion pste whih mixed with 35 ml 80% ethnol (ethnol:wter=80:20) tht ontining 0.1% hydrohlori id into 50-mL entrifuge tue. The smple solution were filtered y ush funnel fter utilizing ultrsoni power (700 W) tretment t room temperture for 30 min. The ove steps were repeted twie. The filtrte of the three times were omined nd entrifuged for 10 min (4 000 r min 1 t 4 C). Finlly, efore the nlysis, the extrt ws filtered y using 0.22 µm reinfored nylon memrne filters. Deionized wter ontining 0.01% hydrohlori id were dded up to 25 ml fter the remin superntnts were omined nd evported under vuum to dryness t 35 C to nlyze the detiled nthoynin nd flvonoids omposition nd their ntioxidnt tivities HPLC-DAD nd HPLC-ESI-MS nlysis The nlysis ws rried out using UHPLC-LTQ Oritrp (Thermofisher Sientifi, USA). The moile phse inluded solvent A (ultrpure wter with queous 0.2% formi id) nd solvent B (100% formi id). The pplied grdient Fig. 1 Onions of three different olors in this study. The left one is yellow ul (w470), the middle one is red ul (w1201), the right one is white ul (628).

3 4 ZHANG Shi-lin et l. Journl of Integrtive Agriulture 2016, 15(0): progrm ws: 35 to 44 min, liner grdient from 5 to 40% B; 45 to 54 min, liner grdient from 40 to 97% B; 55 to 56 min, 97% B isorti; 57 to 116 min, liner grdient 97 5% B. The flow rte ws 0.8 ml min 1 nd the temperture ws 40 C. Anthoynins nd flvonoids were deteted y their sorne t 530, 620 nd 650 nm. The nlytil olumn ws Hypersil C18 reverse (Thermo 2.1 mm 100 mm, 3 μm prtiles). Spetr reording in positive ionistion mode with sns from m/z 200 to ESI soure prmeters were s follows: pillry voltge t 4 kv; neulistion pressure t 35 r nd uxiliry gs pressure t 10 r. 2 µl smples were nlyzed nd the seondry high energy ollision indued dissoition (HCD) reking energy (nomlized ollsion energy) ws Totl nthoynins nd flvonols ontent mesurement The totl nthoynins ontent in onion ws utilized y the ph differentil method of Hung (2009) with some modifitions. 1 ml onion extrt smples were dissolved in 0.2 mol L 1 potssium hloride uffer, ph 1.0, dding the volume up to 25 ml. And nother 1 ml nthoynin extrt ws dissolved in 0.2 mol L 1 sodium ette uffer, ph 4.5, onstnt volume to 25 ml. Smple spetrl sorne mesurements were red t 525 nd 700 nm, respetively. The nthoynins ontent of the diluted smple ws then lulted s follows: nthoynins ontent (mg L 1 )=[(OD 525 OD 700 )ph1.0 (OD 525 OD 700 )ph 4.5] / DF, where, is the reltive moleulr mss of ynidin-3-gluoside; is the molr sorptivity; DF is the dilution ftor; is the ftor to onvert g to mg. The totl flvonols ontent in onion ws mesured y spetrophotometri method (Wng et l. 2014). Rutin ws used s stndrd ompound. Smple spetrl sorne mesurements were red t 510 nm. The flvonol ws lulted from the lirtion urve: R (mg ml 1 )=0.1565OD (R 2 =0.9925) nd flvonols ontent (mg L 1 ) =R DF/m. Where, the DF is 2 500, m is the qulity of the onion smples Totl polyphenol ontent The totl polyphenol ontent (TPC) in the extrt ws determined y Folin-Ciolteu method (Mnsouri et l. 2005). Glli id (GA) ws used s stndrd ompound. Smple spetrl sorne mesurements were red t 750 nm nd the TPC ws lulted from the lirtion urve: TPC (mg ml 1 )=0.6090OD (R 2 =0.9983). The results were expressed s milligrms of GA equivlents per 100 g of FW. 2.6 Antioxidnt pity The DPPH ssy ws dopted sing on the method of Lee (2015) with some modifitions. The smples were diluted to proper onentrtion to ensure the sorne test vlues etween 0.2 to 0.8. The retion solution ws prepred y dissolving 20 mg of DPPH in 100 ml of 80% ethnol solution. Prior to the sorne t 515 nm ws reorded, 20 µl of onion smples were reted with µl of DPPH solution for 2 h in the drk environment t room temperture. GA ws used s stndrd ompound nd the lirtion urve: Y (Svenging rtio)= (GA equivlents ontent ) (R 2 =0.9956). Results were expressed s mg of GA equivlents per 100 g FW. The ABTS + ssy ws performed ording to the referene of Zheng (2011). The ABTS + solution ws diluted to pproprite sorne ( sorne). 10 µl of onion smple ws dded to µl diluted ABTS solution to ret in the drk t room temperture for 10 min, nd the sorne red t 732 nm. GA ws used s stndrd ompound nd the lirtion urve ws Y (Svenging rtio)= (GA equivlents ontent) (R 2 =0.9981). Results were represented s mg of GA equivlents per 100 g FW. FRAP ssy ws used to determine the totl ntioxidnt pity of the smples (Benzie nd Strin 1996) with some modifitions. Briefly, 30 µl of onion smple ws dded to µl of FRAP regent to ret in the drk t room temperture for 15 min nd the sorne t 593 nm ws reorded. GA ws used s stndrd ompound nd the lirtion urve ws Y (Svenging rtio)= (GA equivlents ontent) (R 2 =0.9923). The results were expressed s mg of GA equivlents per 100 g of FW Sttistil nlysis One-wy nlysis of vrine tests (ANOVA) nd ivrite orreltion nlyses were implemented using SPSS19.0. Post ho multiple omprisons were exeuted with the LSD method for ANOVA. The differenes were onsidered to e sttistilly signifint when P<0.05. All experiments were repeted three times. 3. Results 3.1. Totl nthoynins nd flvonols ontent The totl nthoynin ontents determined y the ph differentil method, were (0.75±0.40) mg 100 g 1 in fresh white onion, (9.64±0.30) mg 100 g 1 in fresh yellow onion nd (29.99±1.19) mg 100 g 1 in fresh red onion, respetively. In this study we found tht the nthoynin ontent in onion

4 ZHANG Shi-lin et l. Journl of Integrtive Agriulture 2016, 15(0): were the red>the yellow>the white. The totl flvonoid ontents of onions with three different olors rude extrt were (36.64±3.59) mg 100 g 1, (111.10±5.98) mg 100 g 1 fresh weight in yellow nd red, respetively, wheres there re no flvonols in white onion (Tle 1) Composition of nthoynins nd flvonols The mjor nthoynin nd flvonols omposition of the purified extrt in red nd yellow onions ws further identified y HPLC-ESI-MS, ut we did not reserh those omposition in white onion euse its ontents were extremely low. Delphinidin 3,5-diglyosides, ynidin 3,5-diglyosides, ynidin 3-glyosides nd ynidin 3-(6 -mlonyl)-gluopyrnoside re the four hief nthoynin glyones in onion from the MS dte inluding the pek numer, retention time, moleulr ions nd importnt frgment ions. Queretin nd queretin 3-glyosides re the top two flvonols found in onion (Tle 2, Appendix A). However, the rtio of the pek re from red nd white onions were different tht mens the reltive onstituent ontent of nthoynin nd flvonols etween them hd ovious differene (Appendix B). For exmple, the rtio of ontent of ynidin 3,5-diglyosides in red onion ompred with yellow one ws Wht s more, ynidin 3-(6 -Mlonyl)- gluopyrnoside hs not Tle 1 Three different olor onion nthoynin nd flvonoids ontent (men±se, n=3) Vriety Anthoynins ontent (mg Flvonols ontent 100 g 1 FW) (mg 100 g 1 FW) White onion 0.75± Yellow onion 9.64± ±3.59 Red onion 29.99± ±5.98 een found in yellow onion tht my led to the differene of ul skin of onion Totl polyphenol ontent nd ntioxidnt tivity The results of oth TPC nd ntioxidnt tivity showed in Fig. 2. TPC were (389.69±4.38) (white), (741.84±6.94) (yellow), (622.27±3.91) (red) mg 100 g 1 FW, nd TPC from yellow ws signifintly higher thn other ul skin (P<0.05). By ove mentioned, we relized tht oth nthoynin nd flvonol ontents of white onion were the lowest of ll smples. Three rdil svenging ssys showed similr tendeny with the ontents of nthoynin nd flvonol, nmely the red>the yellow>the white. The red onion presented the mximum ntioxidnt tivity ((82.04±1.98) mg 100 g 1 FW for DPPH ; (175.2±4.35) mg 100 g 1 FW for ABTS + ; (143.37±2.82) mg 100g 1 FW for FRAP, respetively). There is no flvonoids nd low nthoynin ontents in white onions, however, it still showed good ntioxidnt tivity ((33.38±1.14) mg 100 g 1 FW for DPPH ; (66.48±2.24) mg 100 g 1 FW for ABTS + ; (54.36±3.16) mg 100 g 1 FW for FRAP, respetively) Correltion nlysis Correltion oeffiients of nthoynins, flvonols, TPC nd their orresponding ntioxidnt tivity re shown in Tle 3. There ws extremely signifintly positive orreltion etween flvonols nd ntioxidnt tivity s mesured y DPPH (r=0.926), ABTS + (r=0.920), nd FRAP (r=0.807). At the sme time, there ws extremely signifintly positive orreltion (P<0.01) etween nthoynin nd ntioxidnt tivity exept FRAP (r=0.792), while, for the TPC orreltion Tle 2 Anthoynins nd flvonols identified from red nd yellow onions in this study Vriety No. RT (min) Pek re M+ Frgment ions Predited (m/z) formul ppm Pek ssignment Red Ant C 27 H 31 O Delphinidin 3,5-diglyosides onion Ant C 27 H 3 1O Cynidin 3,5-diglyosides Ant C 21 O Cynidin 3-glyosides Ant C 24 H 23 O Cynidin 3-(6 -Mlonyl)-gluopyrnoside Fl , C 15 H 11 O Queretin , , Fl C 21 O Queretin 3-glyosides Yellow onion Ant C 27 H 31 O Delphinidin 3,5-diglyosides Ant C 27 H 31 O Cynidin 3,5-diglyosides Ant C 21 O Cynidin 3-glyosides Fl , C 15 H 11 O 7 Queretin , , Fl C 21 O Queretin 3-glyosides

5 6 ZHANG Shi-lin et l. Journl of Integrtive Agriulture 2016, 15(0): is signifint (P<0.05) in DPPH (r=0.764), ABTS + (r=0.787). The differene my e generted y different speies nd properties. Disussion The white onions ontin less nthoynin ontent thn olored onions tht is onsistent with the study of Herrmnn (1976). However, Even though red onion, its nthoynin onentrtion is lower thn tht of vegetle with sustntil nthoynin pigment, suh s red lef mustrd (Brssi june Coss.) (71.90 mg 100 g 1 FW), purple heding Chinese ge (Brssi rp L. ssp. pekinensis) (60.40 mg 100 g 1 FW), purple-fleshed sweetpotto (Ipomoetts L. Lm) ( mg 100 g 1 FW) (Steed nd Truong 2008; Zhng et l. 2014). This my e resulted from the nthoynins exsiting in eh lyer of onion skin whih only oupy rrely prt of onion fresh weight. Even so, these fts lso suggest tht red onion ould e good soure of nthoynin pigment. The previous study of the ontents of flvonoids in red onion from the northern US (long-dy types) ws higher ompred to our results (mid-dy types) (Yng et l. 2004). In generl, the long-dy type ontined higher mounts of flvonoids thn dy neutrl types nd short dy types (Trmmell nd Peterson 1976). Rodrigues et l. (2010) found out tht s the storge time longer, the ontents of flvonoids will e higher ordingly. Besides, like UV-C irrdition, the nturl levels of flvonoids nd nthoynins in fresh-ut onion slies signifintly inrese (Pérez et l. 2011). However the onion smples of our study were used to the experiment immeditely when hrvested, nd it ould e the reson why its ontent is slightly lower thn other reserhes. In ddition, the reltive referene reserh showed tht lrge prt of the onion flvonoids exsisted outside of the dry skin rther thn the prt of edile fleshy uls. There hve een mny studies on nthoynins nd flvonoids in onion. Compring with the MS informtion, in this pper, we got four types of nthoynins nd two kinds of flvonoids identified in two vrieties (red nd yellow ul). Glii id equivlent (mg FW) TPC ABTS + White Yellow Red DPPH FRAP White Yellow Red 0 White Yellow Red White Yellow Red Fig. 2 Totl polyphenol ontent (TPC) (mg 100 g 1 FW) (mens±se, n=3) nd ntioxidnt pities (DPPH, ABTS + nd FRAP) of eh smple (mg 100 g 1 FW) (mens±se, n=3). Tle 3 The orreltion oeffiients mong nthoynins, flvonols, totl polyphenol ontent (TPC) nd eh ntioxidnt tivity ssy DPPH ABTS + FRAP Anthoynins ** ** * Flvonols ** ** ** TPC * * ** * nd **, orreltion is signifint t the 0.05 nd 0.01 levels (2-tiled), respetively. All nthoynins omponent hs ntioxidnt tivity, however, there were differenes mong them. The ntioxidnt tivities from high to low were ynidin 3-glyosides>ynidin 3,5-diglyosides>ynidin 3-(6 -mlonyl)- gluopyrnoside>delphinidin 3,5-diglyosides (Dnut et l. 2008). Queretin 3-glyosides ntioxidnt tivity ws stronger thn queretin (Kshino et l. 2015). Bsed on the ove resons, we my hoose to et the one whih is rih in

6 ZHANG Shi-lin et l. Journl of Integrtive Agriulture 2016, 15(0): ynidin 3-glyosides nd queretin 3-glyosides, nd py more ttention to improve the ontent of ynidin 3-glyosides nd queretin 3-glyosides in the future reeding work. Compred other similr studies with our results, different olor onions were shown to hve the TPC etween 200 nd mg GAE ml 1 (Chu et l. 2000; Yng et l. 2004). In Chu et l. s (2000) study, the yellow genotype showed higher ontent of TPC thn the red onions. White onion s ntioxidnt tivity indited tht there were likely other polyphenols presene in white onion whih were not deteted in our experiment. Whih mens tht the ntioxidnt tivity my e lso ffeted y other ftors, suh s sori id, phenoli ids, glli id, sulfoompound, et. not involved in this study. Additionlly, Rodrigues et l. (2011) reserhes showed tht onion ntioxidnt tivity levels vried signifintly mong sesons - the ntioxidnt tivity with higher levels when the seson ws very dry nd hot, on the ontrry, it would e lower when the yer with the lowest ir nd soil temperture nd undnt rinfll. Correltion oeffiients of nthoynins, flvonols, TPC, nd their orresponding ntioxidnt tivity re shown ompline with the previous report tht plnts with higher flvonols ontent possess stronger ntioxidnt tivity (Chu et l. 2000). Additionlly, our results showed tht onions, espeilly red nd yellow ones, re potentil resoures of nturl ntioxidnt ompounds. 5. Conlusion In this study, we provided foundtion on the hrteristis of nthoynins nd flvonoids ontents nd omponents of three different olor onions, whih were widely ultivted in middle-est of Chin region. We mesured the ontent of nthoynins nd flvonoids. Pure nthoynins nd flvonoids ompounds were isolted nd nlysed y HPLC-DAD-MS, whih shred the mjor nthoynins nd flvonols omposition. Furthermore, the ntioxidnt properties of three different olor onions were evluted. All types of onions, prtiulrly red onion, hve high ntioxidtive tivities, whih suggests tht they should e good for onsumption of the humn diet nd potentil helth food. Aknowledgements We thnk Dr. Chen Di (College of Life Siene, Nnjing Agriulturl University, Chin) for his help in HPLC-MS nlysis. This study ws finnilly supported y the Speil Reserh of Ntionl Nonprofit Industry (Agriulture), Chin ( ), nd Yngzhou Agriulturl Siene nd Tehnology Reserh Projets, Chin (YZ ). Appendix ssoited with this pper n e ville on Referenes Benzie I F, Strin J J The ferri reduing ility of plsm (FRAP) s mesure of ntioxidnt power: The FRAP ssy. Anlytil Biohemistry, 239, Bonorsi P, Cristi C, Grgiulli C, Leuzzi U Flvonoid gluoside profile of southern Itlin red onion (Allium ep L.). Journl of Agriulturl & Food Chemistry, 53, Brewster J L Onions nd Other Vegetle Alliums. CABI Pulishing,\Wllingford, United Kingdom. pp Bridle P, Timerlke C F Anthoynins s nturl food olours-seleted spets. Food Chemistry, 58, Chu Y H, Chng C L, Hsu H F Flvonoid ontent of severl vegetles nd their ntioxidnt tivity. Journl of the Siene of Food Agriulture, 80, Clifford M N Anthoynins - Nture, ourrene nd dietry urden. Journl of the Siene of Food nd Agriulture, 80, Dnut Z, Wieslw W, Mriusz K P Determintion of the reltive ontriution of queretin nd its gluosides to the ntioxidnt pity of onion y yli voltmmetry nd spetrophotometri methods. Journl of Agriulturl & Food Chemistry, 56, Donner H, Go L, Mzz G Seprtion nd hrteriztion of simple nd mlonylted nthoynins in red onions (Allium ep L.). Food Reserh Interntionl, 30, Fossen T, Andersen Ø M, Øvstedl D O, Pedersen A T, Rknes A Chrteristi nthoynin pttern from onions nd other Allium spp. Journl of Food Siene, 61, Glvno F The hemistry of nthoynins. Funtionl Ingredients. [ ]. mngingyour-usiness/hemistry-nthoynins Herrmnn K Flvonols nd vones in food plnts: A review. Journl of Food Tehnology, 11, Hollmn P C, Arts I C Flvonoids, flvones nd flvnols nture, ourrene nd dietry urden. Journl of the Siene of Food Agriulture, 80, Hou D X Potentil mehnisms of ner hemoprevention y nthoynins. Current Moleulr Mediine, 3, Hung Z, Wng B, Willims P, Rlpheni D P Identifition of nthoynins in musdine grpes with HPLC-ESI-MS. LWT - Food Siene nd Tehnology, 42, Kshino Y, Murot K, Mtsud N, Tomotke M, Hmno T, Muki R Effet of proessed onions on the plsm onentrtion of queretin in rts nd humns. Journl of Food Siene, 80, Kong L, Chi N, Goh T, Chi R Brouillrd, nlysis nd iologil tivities of nthoynins. Phytohemistry, 62,

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