Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.

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Glucosinolates, Glycosidically Bound Volatiles and Antimicrobial Activity of Brassica oleraceae Var. Botrytis, (Soultany Cultivar)
.oha/ed S. Hifnawy+, 0a1a1 .. A1del Sala/2, .oha/ed A. 0a1eh+,3,2 and .ah/oud A. A1oseada3 , +. 3e$art/ent of #har/acognosy, 4aculty of $har/acy, 5airo 6ni7ersity, 5airo,8gy$t, ++&-2. 2. 3e$art/ent of #har/acognosy, 4aculty of #har/acy, Beni-Sweif 6ni7ersity, Beni-Sweif, 8gy$t. 3. 3e$art/ent of #har/acognosy and .edicinal $lants, 4aculty of #har/acy, Nahda 6ni7ersity, Beni-Sweif, 8gy$t. 2 8-/ail of the corres$onding author9 /oha/eda1delatty-!:yahoo.co/; Abstract Background: 8gy$tian cauliflower Brassica oleracea, <. var. Botrytis <. Soultany culti7ar, is an i/$ortant edi1le $lant in .editerranean countries. )nly a few researches were focused on anti/icro1ial acti7ity of its 7olatiles and glucosinolates. Ob ective: =o e7aluate the anti/icro1ial acti7ity of 7olatiles and glucosinolates of 8gy$tian 5auliflower and identify the/ 1y >5?.S and H#<5?.S designs. !aterials and !et"ods: =he se/i7olatile and 7olatile constituents of were e@tracted 1y hydrodistillation fro/ lea7es, ste/s and inflorescences, using a <iAens BNicAerson-ty$e a$$aratus. =he e@tracts fro/ fresh and froCen 7egeta1les were in7estigated 1y >5?.S and H#<5?.S. the 7olatile sa/$les containing glucosinolate degradation $roducts were e7aluated for anti/icro1ial acti7ity using the disc diffusion /ethod. #esults: 4ro/ the fresh lea7es e@tract, a total of 4' co/$ounds were identified, re$resenting '!.,'D of the oil. =he /aEor constituent was found to 1e he@-3"F%-enol "+!.!-D%. 4ro/ fresh disru$ted inflorescence tissues of 8gy$tian cauliflower 4& co/$ounds were detected, re$resenting '3.3,D of the e@tract. Nonacosane and ++-/etho@y 1enC"a%anthracene-,,+2-dione were identified as /aEor constituents of the hydrodistillation $roducts, re$resenting, res$ecti7ely, +,., and !.3+D of the 7olatiles. 4ro/ froCen inflorescence tissues, di/ethyl trisulfide and 1utylated hydro@y toluene were detected as $redo/inant co/$onents re$resenting +&.!! and '.-'D res$ecti7ely. In the latter, he@-3"F%-enol was not found to 1e the /aEor constituent as ha$$ened in fresh lea7es, 1ut re$resenting 3.!!D. 4ro/ fresh ste/ tissues, di/ethyl trisul$hide was detected as /aEor constituent re$resenting a $ercentage "24. -D% /ore than $resent in froCen inflorescence tissues. =wenty two co/$ounds were identified 1y tri$le Guad H#<5?.S. *olatile sa/$les e@$ressed a wide range of growth inhi1ition acti7ity against 1oth >ra/-$ositi7e and >ra/negati7e 1acteria and fungi, showing the highest inhi1itory effects against E .coli and K. pneumonia strains. Conclusion: Hydrodistilled co/$ounds $resent in the ste/s, lea7es and inflorescence tissues of this culti7ar has a highly $ro/ising anti/icro1ial acti7ity. $ey%ords: 5auliflower, Brassica oleraceae, >lucosinolates, >5?.S, H#<5?.S, Anti/icro1ial acti7ity. (ntroduction >lucosinolates "><Ss%, are a class of nitrogen and sulfur-containing natural $roducts distri1uted in +dicotyledonous fa/ilies of the order 5a$$arales "Al-Sheh1aC et al, +'!,; *erAerA et al; 2 '%. .ostly glucosinolates-containing genera are clustered within Brassicaceae, 5a$$araceae and 5aricaceae fa/ilies "4ahey et al., 2 +%.6$on $lant da/age 1y food $rocessing /ethods, such as $hysical distru$tion of $lant cells, chewing and high te/$erature enhance acti7ation of thioglycoside glucohydrolase for hydrolysis and?or ther/al degradation and 7arious 1reaAdown $roducts are for/ed i.e. Isothiocyanates "I=5s%, thiocyanates, nitriles, etc. "4igure +%. "I7ica et al., 2 + %. ><Ss /eta1olites ha7e 1een recogniCed as anti/icro1ial agents for /any decades "4ahey et al., 2 +% and this acti7ity has 1een re$orted to 1e a $art of a 5ruicifers defense against $athogen attacA against such and related strains of hu/an $athogens and fungi "0adulo7ic et al., 2 ++%, and so/e of these /ay 1e $resent in the soil associated with the $lant s$ecies and hence could re$resent direct $lant $athogens "e.g. Aspergillus fumigates ")H>er/an et al., 2 !%. A/ong the degradation $roducts of ><Ss, I=5s ha7e 1een re$orted to $ossess 1road s$ectru/ 1iological acti7ity against 1acterial and fungal $athogens, ne/atodes, insects and weeds "Brown et al., +'',; <in et al., 2 ; .anici et al., 2 ; 0esa et al., +'''%. It is Anown that nitriles are also 1iologically acti7e, 1ut effects are /ore li/ited than are the I=5s "I7ica et al., 2 + %. Just recently )-glycosidically-1ound 7olatile co/$ounds ha7e also 1een re$orted in $lants to the Brassicaceae fa/ily "BloCe7ic et al., 2 ! a,1 I 2 '; .astelic et al., 2 -% rather than ha7e 1een re$orted in /ore than +, $lants 1elonging to & fa/ilies "5rouCet et al., +'''%. >lycosidically -1ound 7olatiles are considered as storage for/s of aro/a co/$ounds that can cause the anti/icro1ial or /edicinal $ro$erties of $lants "5utler et al., +''-; 3e=o//asi et al., +''-%. =hese co/$ounds are ther/ally sta1le, in contrast to intact &'

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Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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><Ss, and are a1le to release 7olatile )-aglycons only 1y acid or enCy/atic hydrolysis "I7ica et al., 2 + %. 0eleased 7olatiles, such as ali$hatic alcohols, $henyl $ro$ane deri7ati7es, 5+3-Noriso$renoids, /onoter$enes and sesGuiter$enes were Anown for their 1iological acti7ities "I7ica et al., 2 + %. Since the end of the si@ties, /uch worA has 1een carried out on the nature of the fla7or 7olatiles of foodstuffs, using a co/1ination of ca$illary gas chro/atogra$hy and /ass s$ectro/etry as well as in7estigation of their anti/icro1ial acti7ities. 5ruciferous 7egeta1les are Anown to contain organic sulfur. =heir characteristic fla7or and odor ha7e 1een attri1uted to 7olatile sulfur containing co/$ounds. .any studies ha7e 1een re$orted, concerning ca11ages, cauliflowers, Brussels s$routs and 1roccoli "Buttery et al., +',-; 3a@en1ichler et al., +',,; .ac<eod et al., +', c; .arAs, et al., +''2; #earson et al., +'!+; *an <angenho7e et al., +''+; Jall1anA et al., +',-%. 5o/$osition of 7olatile constituents has 1een used to identify new 7olatile fla7oring co/$ounds, such as sulfides or I=5s ".aruya/a et al., +', %, to e7aluate the effects of 7ariations in cooAing /ethods ".ac<eod et al., +', a% and to co/$are the 7olatile fractions fro/ 7arious cruciferous 7egeta1les ".ac<eod et al., +', 1%. Studies to i/$ro7e the storage ti/e of cruciferous 7egeta1les ".ac<eod et al., +','% ha7e e/$hasiCed the need for an i/$ro7ed Anowledge of the co/$ounds contri1uting to their fla7or. .uch worA has 1een carried out on raw /aterial. So/e 7egeta1les ha7e 1een in7estigated, to analyCe $reser7ation effects on 7olatile co/$ound co/$osition. ".ac<eod et al., +', % re$orted analytical data on fla7or 7olatile co/$osition of a nu/1er of dehydrated ca11age sa/$les, indicating the $oor nature of the $reser7ed $roduct as a su1stitute for the fresh 7egeta1le. 6sing the sa/e sa/$ling techniGues ".ac<eod et al., +', 1%, the 7olatile constituents of cooAed 7egeta1les were collected and analyCed 1y gas chro/atogra$hy, in order to co/$are the/ with the fresh /aterial. ".ac<eod et al., +','% ha7e also e@a/ined the effect of freeCing and 1lanching on the nature of the 7olatile fla7or co/$onents of Brussels s$routs9 a$$recia1le 7ariations in the che/ical co/$osition of the fla7or e@tracts were o1ser7ed, and $ossi1le reasons for unusual and unacce$ta1ly strong fla7ors of froCen Brussels s$routs were suggested. In /ost of these studies, the 7egeta1les were $laced in 1oiling water. *olatile co/$ounds were waterdistillated and collected at low te/$erature ".aruya/a et al., +', ; .ac<eod et al., +'-!, +', a, +', 1, +', c, +','%, tra$$ed on an adsor$tion tu1e "*an <angenho7e et al., +''+% or e@tracted with sol7ents "3a@en1ichler et al., +',,%. *ery few studies deal with other techniGues, such as hydrodistillation using a <iAensBNicAerson-ty$e a$$aratus "Buttery et al., +',-% or static and dyna/ic heads$ace analysis "Jall1anA et al., +',-%. In each case, sulfides, $olysulfides, thiols, isothiocyanates, nitriles, alcohols, car1onyl co/$ounds "Aetones, aldehydes, and esters%, furans and ter$ene hydrocar1ones were identified in ca11age, cauliflower, Brussels s$routs and 1roccoli. 3i/ethyl trisulfide was $resent as a /aEor aro/a co/$onent in cooAed Brassicaceous 7egeta1les ".aruya/a et al., +', %. .oreo7er, isothiocyanates and nitriles were also found to 1e $redo/inant in Brussels s$routs, whereas aldehydes were the /ost a1undant cauliflower 7olatiles "*an <angenho7e et al., +''+%. 4or ca11age, e7idence 1ased on >5 retention ti/es was o1tained 1y "Buttery et al., +',-%, 2,3,&-trithiahe@ane, di/ethyl sulfide, di/ethyl trisulfide, 2-$henylethyl cyanide and 2- $henylethyl isothiocyanate were identified as i/$ortant aro/a co/$onents. .uch less interest was de7oted to 8gy$tian cauliflower, as this culti7ar has 1een co//ercialiCed for less than +& years. =his 8gy$tian cauliflower 7ariety has a uniGue fla7or and a ty$ical yellowish white, s/all inflorescence re$resenting only 3&- 4 D of total weight of fresh $lant, while the 1y$roducts "ste/s and lea7es% are considered the /aEor $arts. It is not e@$ensi7e as well as other culti7ars grown in 8gy$t such as Snow 1all and Snow crown culti7ars and is a7aila1le anywhere for long ti/e so it is well a$$reciated 1y consu/ers. No s$ectral e7idence has 1een $re7iously o1tained to analyCe 7olatiles isolated fro/ the ste/s. Actually, no studies concerns 7olatile e@traction fro/ 8gy$tian cauliflower. >enerally, no studies included 7olatile e@traction fro/ 1y$roducts e@ce$t carried 1y "*alette et al., 2 3%9 7olatile co/$ounds fro/ raw, ri$ening and froCen florets were e@tracted 1y hydrodistillation co/$ared 1y lea7es and analyCed 1y >5?.S. =he co/$ounds identified in the e@tract and their i/$ortance were descri1ed9 di/ethyl trisulfide, di/ethyl disulfide and he@-3"F%-enol were re$orted as /ain 7olatile constituents in the raw e@tract of florets, whereas di/ethyl trisulfide and di/ethyl disulfide only were $re7alent in ri$ening and froCen florets, 1ut, he@3"F%-enol re$resented indi7idually -+.+D of the oil distillated fro/ lea7es Howe7er, no identification of s$ecific co/$onents was carried out in the ste/s. =he /assi7e increase in $re7alence of /ulti$le drug resistance has slowed down the de7elo$/ent of new synthetic anti/icro1ial drugs, and it necessitated the search for new anti/icro1ials fro/ alternati7e sources. Natural co/$ounds ha7e 1eco/e a source of nu/erous thera$eutic agents. 0ecent $rogress to disco7er drugs fro/ natural sources has resulted in co/$ounds that are 1eing de7elo$ed to treat resistant 1acteria, 7iruses and i//unosu$$ressi7e disorders "A/ghalia et al., 2 '%. #hytoche/icals fro/ /edicinal $lants showing anti/icro1ial acti7ities ha7e the $otential of filling this need, 1ecause their structures are different fro/ those of the /ore studied /icro1ial sources, and therefore their /ode of action are also 7ery liAely to differ. .uch growing interest correlating the $hytoche/ical constituents of a /edicinal $lant with its $har/acological acti7ity

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is needed "#rachayasittiAul et al., 2 !; Nogueira et al., 2 !%. Screening the acti7e co/$ounds fro/ $lants has lead to the disco7ery of new /edicinal drugs which ha7e efficient $rotection and treat/ent roles against 7arious diseases "0oy et al., 2 '%. >lucosinolates were $re7iously re$orted in 5auliflower "Adarsh et al., 2 '; Jon et al., 2 '; 0oCenn et al., +'''% na/ely, sinigrin, glucora$hanin, , >luco$erin, gluco1rassicin etc. as seen in ta1le +. =o our Anowledge, there is no $re7ious re$orts on the 1iological acti7ity of 7olatiles 1elonging to 5auliflower "Botrytis 7ariety, Brassica oleraceae s$ecies%. =hus, the ai/ of this study was to in7estigate the content of these 7olatiles using different /ethods of isolation, na/ely, hydrodistillation and e@traction. .oreo7er, anti/icro1ial acti7ity of the isolates containing glucosinolate degradation $roducts was e7aluated. )' !aterials and met"ods 2.1. General experimental procedures All che/icals that are co//ercially a7aila1le were used as recei7ed "Aldrich, 6SA; .ercA, >er/any; 4luAa, >er/any%, e@ce$t that the sol7ents were $urified 1y distillation. =he >5?.S analyses were $erfor/ed on a HewlettB #acAard -!' N gas chro/atogra$h eGui$$ed with fused silica ca$illary colu/n 3B-& .S "&D $henyl/ethylsilo@ane, 3 / @ .32 //, fil/ thicAness .2& K/, Agilent =echnologies, 6SA% which is cou$led with a &',&B /ass selecti7e detector fro/ the sa/e co/$any as detailed in 2.4. 2.2. Plant materials 8gy$tian cauliflowers, Brassica oleracea <. var. Botrytis <., were o1tained fro/ a local far/, Bayaad el Ara1, Beni-Sweif, 8gy$t%. Botanical identification of the 7egeta1le was authenticated 1y #rof. 3r. A1delhali/ .oha//ed; 4lora de$art/ent, Agricultural /useu/, 5airo, 8gy$t. *oucher s$ecie/en were Ae$t in the Agricultural /useu/. =he studied s$ecies is co//ercialiCed under the na/e LL8gy$tian SoultanyHH culti7ar. All cauliflower sa/$les were cut at the sa/e ti/e. )ne set was analyCed i//ediately; and one set was froCen at 2 o5 until analyCed. 2.3. Extraction of volatiles =his /ethod of $re$aring the oil e@tracts was $rocessed as 1een re$orted in "*alette et al., 2 3% in 0o/anesco cauliflower 1ut with few changes as following9 standard conditions were used for all e@$eri/ents. 4resh lea7es, ste/s and inflorescences "+Ag% were cut into s/all $ieces and $laced in a 2 liters flasA with +.& liter of water. =he /i@ture was su1Eected to hydrodistillation using a <iAensBNiAerson ty$e a$$aratus. =he 7olatile fraction was e@tracted under at/os$heric $ressure during 4 h, using diethyl ether "& /l% as e@tracting sol7ent. =his e@tract was dried o7er anhydrous sodiu/ sul$hate and carefully concentrated to s/all 7olu/e "~ .& /l%, with a rotary e7a$orator at at/os$heric $ressure. 4urther, this oil was stored at low te/$erature $rior to analysis. =he froCen inflorescences were hydrodistilled, following the /ethod $re7iously descri1ed. =he froCen 7egeta1les were first thawed using a /icrowa7e o7en during + /in "defroCen /ode%. Before analysis, the e@tracts were treated as indicated a1o7e. 2. . !apillary G!"mass spectral analysis 8ach concentrated e@tract sa/$le was analyCed 1y >5?.S, using HewlettB #acAard-Agilent -!' N syste/, with a 3B-& "&D $henyl/ethylsilo@ane, 3 / @ .32 // fused silica ca$illary colu/n; fil/ thicAness, .2& K/% which is cou$led with a &',&B /ass selecti7e detector. >5 o7en initial te/$erature was 4 o5 and was $rogra//ed to 2! o5 at a rate of 2o5 ? /in., and finally held at 2! o5 during +2 /in. )$erating conditions of the >5 were as follows9 heliu/ was used as carrier gas "+ /l?/in%; the te/$erature of inEector and detector was 2& o5 and 2! o5, res$ecti7ely; the 7olu/e inEected was +Kl "diethyl ether solution% in s$litless /ode. 0etention indices were deter/ined co/$ared co/$ared with "Ada/s et al., 2 '; Mo7atNs inde@ $hero1ase., 2 +2; *alette et al., 2 3 I 2 &; 4ati/a et al., 2 + I 2 '; Al->endy et al., 2 + ; I7ica et al., 2 + ; Nico et al., 2 +2% as references. =he /ass s$ectra were $erfor/ed at , e* of the /ass range of & B' . No re$licates were $erfor/ed for any sa/$le. =he 7alues were deter/ined for each co/$onent identified "=a1le +%. 2.#. $P%!&'A'&().().()* 2.#.1. !+emicals* .ethanol, acetonitrile H#<5 gradient grade were o1tained fro/ BaAer "J.=. BaAer, 3e7enter, =he Netherlands%. 4or/ic acid was $urchased fro/ Sig/aBAldrichB4luAa ".adrid, S$ain%. 2.#.2. )ample treatment* 5auliOower inflorescences, lea7es and ste/s of B. oleracea were $acAed in #8?alu/inu/ 1ags under 7acuu/, froCen at -! o5 and analysed within /onths. Sa/$le e@traction was carried out following "Song et al., 2 ,%, with /odiPcations to taAe into account the Oa7onoids and the sina$ic acid deri7ati7es. 4roCen cauliOower sa/$les were /inced in a 1lender /i@er 0o1ot-cou$e Bli@er 3 "0o1ot 5ou$e "6M% <td., Isleworth 6M%. An aliGuot of +.& g of froCen sa/$le was $laced in centrifuge tu1e, followed 1y ,.& /l of cold "4o5% /ethanol to /ini/iCe enCy/atic acti7ities. Sa/$les were then $ut in a ther/ostatic water 1ath ".e//ert >/1HQ 5o., M>, Schwa1ach, >er/any% and /aintained at , o5 for +& /in. 8@tracts were refrigerated in an ice water 1ath and centrifuged at + , r$/ for + /in at 4o5 in a BecA/an J2-.5 centrifuge "BecA/an Instru/ents IN5., #alo

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Alto, 5A, 6SA%. An aliGuot of 2 /< of the clean su$ernatant was e7a$orated to dryness with nitrogen and reconstituted with + /< of /o1ile $hase A "+ D /ethanol in .2 D for/ic acid%, Pltered through a #=48 .2 K/ Plter and inEected into the H#<5-.S?.S?.S syste/ under the conditions re$orted. 2.#.3. $P%!"mass spectral analysis =he chro/atogra$hic syste/ consisted of a 3ione@ 6lti/ate 3 H#<5 "Bre/en, >er/any% co/$osed of a Guaternary $u/$ with an on line degasser, a ther/ostated colu/n co/$art/ent, a $hotodiode array detector "3A3%, an auto sa/$ler, and 5hro/elon software. =he H#<5 se$aration was $erfor/ed on Fo1ra@ SB-5+! colu/n "+& //R4.- //, +.! K/, Agilent 5o/$any, 6SA%, at a Oow rate of .! /l?/in. =he colu/n o7en te/$erature was set at 3 o5 and the inEected 7olu/e was &, + and + K< of lea7es, inflorescences and ste/s res$ecti7ely. .o1ile $hase consisted of two sol7ents, "A% /ethanol and "B% .2 D for/ic acid. Se$aration of co/$ounds was carried out with gradient elution $rofile9 /in, A9 B + 9' ; 3- /in, A9 B , 93 ; & /in, A9 B + 9 ; - /in. =he H#<5B.S syste/ consisted of electros$ray ioniCation "8SI% interfaced BruAer 3altoniA 8sGuire-<5 A/aCon S< Ion =ra$ .ass s$ectro/eter "Bre/en, >er/any% and 3ione@ 6lti/ate 3 ">er/any%, /entioned a1o7e. =he ioniCation $ara/eters were as follows9 $ositi7e ion /ode; ca$illary 7oltage 4 *, end $late 7oltage S& *; ne1uliCing gas of nitrogen at& $.s.i.; drying gas of + l?/in nitrogen at 3& T5. .ass analyCer scanned fro/ +& to + u. =he .S?.S?.S s$ectra were recorded in auto-.S?.S?.S /ode. =he frag/entation a/$litude was set to +. *. .S3 data were acGuired in negati7e /ode. 2.,. -n vitro antimicro.ial activity 2.,.1. /est microorganisms Nine 1acterial strains and four fungal strains used in the $resent study. =he 1acteria used were )tap+ylococcus aureus "05.B + 2,%, )tap+ylococcus epidermidis "05.B + 24% )treptococcus pyogenes "05.B + +&% as >ra/-$ositi7e and >ra/-negati7e 0eisseria gonorr+oeae "05.B + ,-%, Proteous vulgaris "05.B + !&%, Kle.siella pneumoniae "05.B + '3%, )+igella flexneri "05.B + &42%, Pseudomonas aeruginosa "05.B + 43% and Esc+eric+ia coli "05.B + &-%. =he fungal strains used were Aspergillus fumigates ""05.B 2&-4%, !andida al.icans ""05.B & 3&%, Geotricum candidum "05.B & '-% and /ric+op+yton mentagrop+ytes "05.B '2&%. =hey were o1tained fro/ the 0egional 5enter of .ycology and Biotechnology Anti/icro1ial 6nite "05.B%, 5airo, 8gy$t. 2.,.2. !ulture media and anti.iotics for antimicro.ial assay* Brain Heart Infusion as liGuid and solid /edia "Hi.edia%, .ueller- Hinton agar "Hi.edia% and #otato 3e@trose Agar /ediu/ "Hi.edia% were used. A/$icilin ()@oid, 6M) and >enta/ycin ()@oid, 6M) were used as standard anti1acterial agents and A/$hotricin B "Sig/a 5he/ical 5o., St. <ouis, .o.% was used as a standard antifungal agent. 2.,.3. 'etermination of t+e antimicro.ial activity* =he anti/icro1ial acti7ity of oil e@tracti7es of inflorescences, lea7es and ste/s of 8gy$tian cauliflowers, Brassica oleracea was screened 1y the agar disc diffusion /ethod descri1ed 1y Bauer et al., +'--, N55<S, +''3 and Sahoo et al., 2 - with slight /odification. =he 1acterial cultures were grown in Brain Heart Infusion 1roth at 3,*5. After - h of growth, + Ul of each /icroorganis/ at a concentration of +@+ -cells?/l was inoculated on the surface of .ueller-Hinton agar $lates for 1acteria and #otato 3e@trose Agar $lates for fungi. 3.S) with a concentration u$ to 2D was used to dissol7e the oil e@tracts. 4ilter $a$er discs "- // in dia/eter% saturated with 2 U< of the tested essential oils or 3.S) "sol7ent control% were $laced on the surface of the inoculated $lates. =o e7aluate the efficiency of the /ethodology; & Ul of each essential oil was inserted si/ultaneously in a hole /ade in new $lates. =he $lates were incu1ated at 3, T5 for 24 h. =he dia/eter of the inhi1ition Cone was /easured in /illi/eter, and was recorded as /ean V S3 of a tri$licate e@$eri/ent. A/$icillin "+ Ug%, genta/icin "+ Ug% discs for 1acteria, and a/$hotericin B "& Ug% for fungi were used as $ositi7e standard. 5ultured s$ecies $roducing halos eGual to or greater than , // were considered susce$ti1le to the tested essential oil. =he results are shown in ta1le 3. +' #esults and discussion 3.1. !+emical composition of volatiles* *olatile co/$ounds and?or constituents that are $resent in trace a/ounts /ay contri1ute significantly to the characteristic fla7or and aro/a of fresh, ri$ening or cooAed foods. =his in7estigation was $erfor/ed to identify the che/ical structure of i/$ortant aro/a co/$onents in the 8gy$tian cauliflower WSoultany culti7arHH. =he 7olatile fraction was in7estigated 1y a >5?.S co/1ination. All the constituents were identified 1y co/$aring their /ass s$ectra with those stored in .S data1ases "Jiley ,N; .ass 4inder , Jilly, Nist &; .ass 4inder <i1raries%, with literature data "Ada/s, 2 '; *alette, 2 3%, and with ho/e-/ade /ass s$ectra li1raries "1uilt u$ fro/ $ure su1stances and .S literature data% in addition to retention indices concurrently inEected under the sa/e conditions. Identification was confir/ed 1y co/$arison of their s$ectra

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frag/entation $attern with >5 data $re7iously $u1lished "3a7ies, +'' ; 8S) 2 , *alette, 2 3%. =he co/$onents of the 8gy$tian cauliflower e@tracts, the $ercentage of each constituent and the retention indices are gi7en in =a1le +. 5o/$ounds are listed in order of their elution on the 3#-& colu/n. =he chro/atogra$hic $rofile of fresh lea7es oil re7eals that it contains &+ constituents, re$resenting '!.,'D of the e@tract. =he studied sa/$le was do/inated 1y the unsaturated alcohol fraction that a/ounted to 3 .--D of the oil. In $articular, he@-3"F%-enol "+!.!-D% was found to 1e the /aEor constituent of the e@tract. )ther /inor co/$ounds were o1ser7ed, such as di/ethyl trisulfide " .,,D%, anethol"F% "3.'3D%, 3"F%-he@en-+-ol-acetate "+.3+D% and 1utylated hydro@y toluene "3.''D% , although their $ro$ortions were insignificant co/$ared with the alcohols. =he sa/e e@$eri/ent was conducted with fresh inflorescences. A total of 4& 7olatile constituents were detected in the sa/$le oil, re$resenting '3.3,D of the e@tract. =he 7olatiles of inflorescences were a1undant in alAanes and aro/atic Aetonic co/$ounds, /ore e@actly in nonacosane "+,.,D%, ++-/etho@y1enCXaY-anthracene,,+2-dione "!.3+D% and 1utylated hydro@y toluene "2.,,D%. It is noteworthy that 3 sulfur co/$ounds " .+'B .,4D% were also identified, according to the literature, and their /ass s$ectra data. In addition he@-3"F%-enol was not detected a/ong the 7olatile constituents of the studied sa/$le co/$ared to the lea7es "+!.!-D%. 5o/$aring the chro/atogra$hic $rofiles o1tained with fresh lea7es and fresh inflorescence, an interesting fact is noteworthy9 the alAane and aro/atic fractions were $redo/inant in 8gy$tian cauliflower inflorescence, whereas unsaturated alcohols are the /aEor co/$onents of the lea7es of this 7egeta1le. =his has not already 1een re$orted 1y .ac<eod et al. "+', a, +', 1%9 the sulfur 7olatiles are rather $roduced 1y the young $art of the ca11age whereas the older lea7es li1erate a greater $ro$ortion of alcohols, aldehydes and Aetones or 1y *alette, 2 39 the sul$hide fraction is $redo/inant in 0o/anesco cauliflower inflorescences, whereas unsaturated alcohols were the /aEor co/$onents of the lea7es of this 7egeta1le. .ost of the 7olatiles li1erated fro/ the cauliflower are $roduced 1y the inflorescence rather than the outer lea7es. 4roCen inflorescence was in7estigated under the sa/e conditions. A total of 4& co/$ounds were detected, re$resenting ''D of the 7olatiles. =his sa/$le was do/inated 1y the sulfur and the $henolic fractions, re$resenting, res$ecti7ely, 24.4, and '.-'D of the oil. 3i/ethyl trisulfide has already 1een re$orted as the /aEor aro/a co/$onent in cooAed Brassicaceous 7egeta1les 1y .aruya/a "+', %. =he fla7our $rofile of fresh cauliflower is Guite different fro/ that of the $reser7ed 7egeta1le. =he decrease of unsaturated alcohols, o1ser7ed in froCen 7egeta1les, is co/$ensated 1y a huge increase in $olysulfide 7olatiles. =he increased a/ount of such co/$ounds $ro1a1ly contri1utes to the characteristic cauliflower fla7our, distinguishing the fresh 7egeta1le fro/ the froCen one. An i/$ortant 7ariation concerns the ratio of aldehydes $roduced. It is considera1ly reduced in fresh cauliflower, whereas it increased in froCen one. =he 7olatile constituents of fresh ste/s were identified under the sa/e conditions. A total of 4& co/$onents were detected, re$resenting '3.'D of the 7olatiles. =his sa/$le was do/inated 1y the sulfur fraction re$resenting 3 .-2D of the 7olatiles. As froCen inflorescences, di/ethyl trisul$hide was considered to 1e the /aEor constituent, re$resenting 24. -D of the oil. =his $ercentage is /ore than $resent in froCen inflorescence "+&.!!D% whereas the $henolic fraction is reduced in ste/ tissues. >lucosinolate deco/$osition leads to the for/ation of isothiocyanates or cyanides, .ac<eod "+', c% suggested that glucosinolates are deco/$osed enCy/atically, to gi7e isothiocyanates and ther/ally, to afford the cyanides. =he decrease in isothiocyanate ratio is $ro1a1ly due to the inacti7ation of the enCy/e /yrosinase during the $reser7ation stage. =he increased a/ount of cyanides is /ost liAely the result of a ther/al deco/$osition of the sinigrin during the $rocessing. =he /ain differences o1ser7ed 1etween fresh and froCen 8gy$tian cauliflower are not in good agree/ent with those already descri1ed 1y .ac<eod et al., "+', 1% in B. oleracea <. 7ar. ge//ifera <. and 7ar. Botrytis < and 1y *allete, 2 3 in 0o/anesco cauliflower, in that the cyanides and isothiocyanates were identified only in fresh inflorescences and ste/s of 8gy$tian cauliflower. Allyl isothiocyanate, 4"/ethylthio% 1utyl cyanide, 2-$henylethyl cyanide, 1utane-+- isothiocyanate and 2-$henylethyl isothiocyanate were identified in fresh ste/s, whereas 4-"/ethylthio% 1utyl cyanide alone was identified in fresh inflorescences. =hese co/$onents were not identified a1solutely in lea7es and inflorescences of 8gy$tian cauliflower with unAnown reasons. 3.2. $P%!"mass spectral analysis )n the 1asis of their tri$le Guad frag/entation ions and 6**is s$ectra, H#<5-3A3-.S?.S?.s /ethod was used for deter/ination of the /aEor glucosinolates, $henol co/$ounds and $hytoale@ines in B. oleraceae sa/$les and e@traction $rocedure 1y an H#<5 se$aration with tri1le detection "3A3 and .S?.S?.S% were o$ti/iCed to allow a suita1le e7aluation of these nutritionally rele7ant co/$ounds in cauliflower sa/$les. =wel7e glucosinolates were identified as Sinigrin "2-#ro$enyl glucosinolate% X&Y, >lucona$in "But-3-enyl glucosinolate% X)Y, #rogoitrin "2-Hydro@y1ut-3-enyl glucosinolate% X+Y, >lucoerucin "4.ethylthio1utyl glucosinolate% X,Y, >lucoi1erin "3-.ethylsulfinyl$ro$yl glucosinolate% X-Y, >lucora$hanin "4.ethylsulfinyl1utyl glucosinolate% X.Y, >lucotro$aeolin "BenCyl glucosinolate% X/Y, >luco1rassicin "Indol-3-

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yl/ethyl glucosinolate% X0Y, Neo-gluco1rassicin "+-.etho@yindol-3-yl/ethyl glucosinolate% X1Y, 4.etho@ygluco1rassicin "4-.etho@yindol-3-yl/ethyl glucosinolate% X&2Y, >lucoi1er7erin "2-thia$entayl glucosinolate% X&&Y and n-he@yl glucosinolate "/ethyl$entyl glucosinolate% X&)Y "4igure 3A%. In addition, si@ fla7onoids were also identified, na/ely Mae/$ferol-3-diglucoside-,-glucoside X&+Y, Mae/$ferol-3-triglucoside,-glucoside X&4Y, Mae/$ferol -,-o-glucoside X&-Y, Mae/$ferol -3-o-"sina$oyl%-so$horoside-,-o-glucoside X&-Y, 0utin X&,Y and A$igenin X&0Y "4igure 3B%.. In a si/ilar way, three #henolic acids and?or glycosides were identified, na/ely +-sina$oyl-2-feruloyldiglucoside X&'Y, +,2,2N-trisina$oyldiglucoside X)2Y and chlorogenic acid X)&Y "4igure 35% 1eside one $hytoale@ine na/ely 5aulile@in 5 "+-/etho@yindol-3-ylacetonitrile% X))Y "4igure 3 3%. =he concentrations of glucosinolates, $henolic co/$ounds and $hytoale@ines found were deter/ined and found to 1e 7aria1le slightly with $re7iously $u1lished data of Song et al., 2 & and =ian et al., 2 &. =his slight 7ariation can 1e attri1uted to the en7iron/ental conditions and the conseGuence of the different origin of our sa/$les' =he glucosinolate $rofile of B. oleracea ste/s was /arAed 1y high relati7e le7els of glucoi1erin "2.2D%, sinigrin "+.2D%, glucoi1er7erin "3.+D%, glucona$in "3.2D%, glucoerucin "+.2D%, gluco1rassicin "+.3D%, 4/etho@y-gluco1rassicin "3.2D%, neogluco1rassicin ",.3D% and n-he@yl glucosinolate "3.!D% whilst the lea7es were richer in neogluco1rassicin "-.,D% and n-he@yl glucosinolate "2. D% and /ainly rich in fla7onoids Mae/$ferol-3-diglucoside-,-glucoside "2.'D%, Mae/$ferol-3-triglucoside-,-glucoside "2.,D%, Mae/$ferol -,-oglucoside ",.4D%, Mae/$ferol -3-o-"sina$oyl%-so$horoside-,-o-glucoside "+.2D% and 0utin "2.2D% /eanwhile, inflorescence were $oor in glucosinolates and contained "+.4D% a$igenin fla7onoid and this in agree/ent with $re7iously re$orted studies "Branca et al., 2 2; *olden et al., 2 '%. =he results are shown in ta1le 2 and figures 2, 3. 3.3. Antimicro.ial activity =he anti/icro1ial acti7ity of B. oleraceae var..otrytis, 8gy$tian grou$, Soultany culti7ar 7olatile sa/$les, o1tained 1y hydrodistillation followed 1y e@traction e7aluated 1y /easuring the dia/eters of the inhi1ition Cones for all sa/$les at the 7olu/e of 2 Kl $er disc. =he results $resented in ta1le 4 show that the 7olatile sa/$les isolated 1y hydrodistillation followed 1y e@traction e@hi1ited anti/icro1ial acti7ity, 1ut not against all /icroorganis/s tested. =hese 7olatile sa/$les show the highest inhi1itory effects against E. coli and K. pneumonia, which are well Anown as naturally resistant to /any anti/icro1ial agents. =he 7olatile sa/$le of ste/s showed the strongest anti/icro1ial acti7ity against E. coli with a /ean Cone of inhi1ition dia/eter of +'.,V+.3, //. All other tested >ra/-negati7e 1acteria also dis$layed 7aria1le degrees of susce$ti1ility against all 7olatile sa/$les, with a range of ++.'V .!3- +-.,V+.+& // Cone of inhi1ition dia/eter with so/e e@ce$tions9 P. aeruginosa did not dis$lay any degree of susce$ti1ility against all 7olatile sa/$les; neither inflorescences nor lea7es 7olatile sa/$les e@hi1ited anti/icro1ial acti7ity against P. vulgaris; lea7es 7olatile oil indi7idually had no anti/icro1ial acti7ity against 0. gonorr+oeae. All tested >ra/-$ositi7e 1acteria were also sensiti7e against 7olatile sa/$les isolated 1y hydrodistillation followed 1y e@traction e@ce$t for lea7es sa/$le with Cone of inhi1ition dia/eter ranging fro/ +3.!V+.33 to +'.2V+.2' //. >ra/-$ositi7e ). epidermidis was the /ost sensiti7e to 7olatile sa/$le of ste/s containing the highest $ercentage of sulfur co/$ounds o1tained 1y hydroditillation of fresh $lant /aterial, with a /ean Cone of inhi1ition of +'.2V+.2' // "Bauer et al., +'--% was in a good agree/ent with these results u$on using 1enCoyl $ero@ide-sulfur crea/ for acne 7ulgaris against ). epidermidis strain. =hese 7olatile sa/$les were also found to inhi1it the growth of fungal organis/s "!. al.icans, G. candidum, /. mentagrop+ytes and A. fumigatus% with Cone of inhi1ition dia/eter 7alues ranging fro/ '.3V .&2 to +'.,V+.2' //. Since B. oleraceae s$ecies, Botrytis variety 7olatile isolates contain high $ercentages of glucosinolates and glucosinolate degradation $roducts, as also do /any $lants 1elonging to fa/ily Brassicaceae, this in7estigation confir/s the i/$ortance of studying the correlation 1etween che/ical co/$osition of 7olatiles and anti/icro1ial acti7ity. ,' Conclusion =his is the first re$ort concerning the hydrodistilled co/$ounds $resent in the ste/s, lea7es and inflorescence tissues of this culti7ar. =o our Anowledge, there are no $re7ious re$orts on the 1iological acti7ity of 7olatiles 1elonging to 5auliflower "B. oleraceae s$ecies, Botrytis variety%. =he $re7ious re$orts showed anti/icro1ial acti7ities of 7olatile isolates o1tained fro/ other 7arieties of Brassica oleraceae s$ecies such as re$orted 1y "Sousa et al., 2 !%, or s$ecies of different genus that 1elong to the sa/e fa/ily, such as Eruca sativa "Mhoo1chandani et al., 2 + %, 1asa.ia 2aponica "Shin et al., 2 4%, Pseudocytisus integrifolius "Bendi/erad et al., 2 &%, 3ap+anus sativus "Bee7i et al., 2 '; 0ani et al., 2 !% and Erysimum c+orintum "Al->endy et al., 2 + %. 0egardless of the isolation /ethod "hydrodistillation and e@traction% and differences in 7olatiles

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co/$osition, all isolates e@hi1ited different degrees of anti/icro1ial acti7ity against >ra/-$ositi7e and >ra/negati7e 1acteria and fungi. Also, there were no significant differences in anti/icro1ial acti7ity a/ong all 7olatile sa/$les against /ost /icroorganis/s tested. An e@ce$tion was noticed with lea7es 7olatiles which were inacti7e against /ost /icroorganis/s tested. >lucosinolate degradation $roducts, isothiocyanates and nitriles /ay 1e res$onsi1le for the o1ser7ed anti/icro1ial $otency, as they constitute /ost of the 7olatile sa/$les of B. oleraceae. Ackno%ledgment =he authors would liAe to thanA #har/acognosy 3e$art/ent, 5airo 6ni7ersity for $ro7iding facilities and eGui$/ent for the research. =he authors inde1t to 0egional 5enter for .ycology and Biotechnology Anti/icro1ial unit "05.B%, 5airo, 8gy$t for great hel$ in conduction of anti/icro1ial acti7ities. #eferences Ada/s 0#. Identification of essential oil co/$onents 1y gas chro/atogra$hy?/ass s$ectrosco$y. 5arol Strea/, I<9 Allured #u1l. 5or$; 2 '. Al->endy AA, 8l-gindi )3, HafeC AlS, Ateya A.. >lucosinolates, 7olatile constituents and 1iological acti7ities of Erysimum corint+ium Bioss, 4Brassicaceae5. 4ood che/. 2 + ; ++!9 &+'-24. Al-Sheh1aC IA, Al-Sha//ary MI. 3istri1ution and che/ota@ono/ic significance of glucosinolates in certain .iddle-eastern !ruciferae. Bioche/ Syst 8colo J.+'!,; +&9 &&'B-'. A/ghalia 8, Nagi AA, Sha/sudin .N, 0adu S, 0osli 0, Neela *, 0ahi/ 0A, et al. .ulti$le@ #50 assays for the detection of clinically rele7ant anti1iotic resistance genes in )tap+ylococcus aureus isolated fro/ .alaysian hos$itals. 0es J Bio Sci. 2 '; 4"4%9 444-!. Bauer AJ, Mir1y .3M, Sherris J5, =urcA .. Anti1iotic susce$ti1ility testing 1y standard single disc diffusion /ethod. A/ J 5lin #athol. +'--; 4'3--. Bee7i SS, .anga/oori <N, Ana1rolu N. 5o/$arati7e acti7ity against $athogenic 1acteria of the root, ste/, and leaf of 3ap+anus sativus grown in India. Jorld J .icro1iol Biotechn. 2 '; 2&9 4-&B,3. Bendi/erad N, =ale1 Bendia1 SA, Bena1adEi AB, 4ernandeC (, *alette <, <iCCani-5u7elier <, et al. 5o/$osition and anti1acterial acti7ity of Pseudocytisus integrifolius "Salis1.% essential oil fro/ Algeria. J Agric 4ood 5he/. 2 &; &39 2'4,B&2. BlaCZe7ic I, .astelic J. 4ree and 1ound 7olatiles of garlic /ustard "Alliaria petiolata%. 5roatica 5he/ Acta. 2 !1; !+9 - ,B+3. BlaCZe7ic I, .astelic J. 4ree and 1ound 7olatiles of rocAet "Eruca sativa .ill.%. 4la7our 4rag J. 2 !a; 239 2,!B !&. BlaCZe7ic I, .astelic J. >lucosinolate degradation $roducts and other 1ound and free 7olatiles in the lea7es and roots of radish "3ap+anus sativus <.%. 4ood 5he/. 2 '; ++39 '-B+ 2. BlaCZe7ic I, 0adonic A, .astelic J, FeAic ., SAoci1u[ic ., .ara7ic A, et al. >lucosinolates, glycosidically 1ound 7olatiles and anti/icro1ial acti7ity of Aurinia sinuate 4Brassicaceae5. 4ood che/. 2 + ; +2+9 + 2 -!. Branca 4, <i >, >oyal S and \uiros 54. Sur7ey of ali$hatic glucosinolates in Sicilian wild and culti7ated Brassicaceae. #hytoche/istry, 2 2; &', ,+,B,24. Brown #3, .orra .J. 5ontrol of soil-1orne $lant $ests using glucosinolate-containing $lants. Ad7 Agron. +'',; -+9 +-,B23+. Buttery 0>, >uadagni 3>, <ing <5, Seifert 0., <i$ton J. Additional 7olatile co/$onents of ca11age, 1roccoli, and cauliflower. J Agric 4ood 5he/. +',-; 24"4%9 !2'B32. 5rouCet J, 5hassagne 3. >lycosidically 1ound 7olatiles in $lants. in9 IAan 0, editor. Naturally occurring glycosides. New ]orA9 John Jiley and Sons; +'''. #. 22&B,4. 5utler H>, Hill 0A, Jard B>, 0ohitha H, Stewart A. Anti/icro1ial, insecticidal and /edicinal $ro$erties of natural $roducts, fla7ors and fragrances. in9 =aAeoAa >0, =eranishi 0, Jillia/s #J, Mo1ayashi A, editors. Biotechnology for i/$ro7ed foods and fla7ors . Jashington 359 J A/ 5he/ Soc; +''-. 3a7ies NJ. >as chro/atogra$hic retention inde@es of /onoter$enes and sesGuiter$enes on /ethyl silicone and car1owa@ 2 . $hases. J 5hro/atogr. +'' ; & 39 +B24. 3a@en1ichler .8, *an 8tten 5H, S$encer >4. >lucosinolates and deri7ed $roducts in cruciferous 7egeta1les. Identification of organic nitriles fro/ ca11age. J Agric 4ood 5he/. +',,; 2&"+%9 +2+B4. 3e =o//asi N, 0astrelli <, 5u/anda J, S$eranCa >, #iCCa 5. Aryl and triter$enic glycosides fro/ (argyricarpus setosus. #hytoche/. +''-; 429 +-3B,. 8S), =he co/$lete data1ase of essential oils. =he Netherlands9 Boelens Aro/a 5he/ Inf Ser7; 2 . 4ahey JJ, Falc/ann A=, =alalay #. =he che/ical di7ersity and distri1ution of glucosinolates and isothiocyanates a/ong $lants. #hytoche/. 2 +; &-9 &B&+ 4ernandeC 4, #eriera 3., de #inho #>, *alentao #, #eriera JA, Bento A, Andrade #B, et al. .eta1olic fate of

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dietary 7olatile co/$ounds in Pieris .rassicae. .icroche/ J. 2 '; '39 ''B+ '. 4ernandeC 4, #eriera 3., de #inho #>, *alentao #, #eriera JA, Bento A, Andrade #B, et al. Heads$ace solid$hase /icroe@traction and gas chro/atogra$hy?ion tra$-/ass s$ectro/etry a$$lied to a li7ing syste/9 Pieris .rassica fed with Aale. 4ood che/. 2 + ; ++'9 +-!+-'3. Mhoo1chandani ., )Eeswi BM, >anesh N, Sri7asta7a .., >a11anini S, .atera 0. Anti/icro1ial $ro$erties and analytical $rofile of traditional Eruca sativa seed oil9 5o/$arison with 7arious aerial and root $lant e@tracts. 4ood 5he/. 2 + ; +2 9 2+,B24. Mo7ats retention inde@ of organic co/$ounds, New co7er retention inde@ of /ore than +3 con$ounds in 32 records Xho/e $age on the enternetY. #uNer9 =he $hero1ase9 8@tensi7e 3ata1ase of Insect #hero/ones and Se/ioche/icals, Inc.; c2 3-+2 Xcited 2 +2 .ay ++Y. A7aila1le fro/9 htt$9??www.$hero1ase.co/?data1ase?Ao7ats?Ao7ats-inde@.$h$. <in 5., Mi/ J., 3u J(, Jei 5I. Bactericidal acti7ity of isothiocyanate against $athogens on fresh $roduce. J 4ood #rot. 2 ; -39 2&B3 . .ac<eod AJ, .ac<eod >. 8ffects of 7ariations in cooAing /ethods on the fla7or 7olatiles of ca11age. J 4ood Sci. +', a; 3&9 ,44B& . .ac<eod AJ, .ac<eod >. 4la7or 7olatiles of so/e cooAed 7egeta1les. J 4ood Sci. +', 1; 3&9 ,34-!. .ac<eod AJ, .ac<eod >. *olatiles of cooAed ca11age. J Sci 4ood Agric. +'-!; +'"&%9 2,3B,. .ac<eod AJ, #iAA H8. *olatile fla7or co/$onents of fresh and $reser7ed 1russels s$routs grown at different cro$ s$acings. J 4ood Sci. +','; 449 ++!3B&, ++' . .ac<eod J, .ac<eod >. =he fla7or 7olatiles of dehydrated ca11age. J 4ood Sci. +', c; 3&9 ,3'-43. .anici <., <aCCeri <, BaruCCi >, <eoni ), >alletti S, #al/ieri S. Su$$ressi7e acti7ity of so/e glucosinolate enCy/e degradation $roducts on Pyt+ium irregular and 3+i6octonia solani in sterile soil. #est .anag Sci. 2 ; &-9 '2+B-. .arAs HS, J Hilson, A, <eichtweis H.5, Stoewsand >.S. S-.ethylcysteine sulfo@ide in Brasssica 7egeta1les and for/ation of /ethyl/ethanethiosulfinate fro/ Brussels s$routs. J Agric 4ood 5he/. +''2; 4 "++%9 2 '!B + +. .aruya/a 4=. Identification of di/ethyl trisulfide as a /aEor aro/a co/$onent of cooAed Brassicaceous 7egeta1les. J 4ood Sci. +', ; 3&9 &4 B3. .astelic J, BlaCZe7ic I, JerAo7ic I. 4ree and 1ound sul$hur containing and other 7olatile co/$ounds fro/ e7ergreen candytuft "-.eriss empervirens <.%. 5roatica 5he/ Acta. 2 -; ,'9 &'+B,. .inard 0, <arue J, Silue 3, =hou7enot 3. >lucosinolates in cauliflower as 1ioche/ical /arAers for resistance against downy /ildew. #hytoche/. +'''; &29 2'-3&. National 5o//ittee for 5linical <a1oratory Standards "N55<S%. #erfor/ance standards for anti/icro1ial susce$ti1ility testing. 8le7enth infor/ational su$$le/ent. 2 3; 3ocu/ent .9 + -S++. Nogueira J5, 3iniC .de 4, <i/a 8). -n vitro anti/icro1ial acti7ity of $lants of acute otitis e@terna. BraC J )torhinolaryngol. 2 !; ,4"+%9++!B24. )H>or/an 5., 4uller H=, 3yer #S. 3isco7ery of a se@ual cycle in the o$$ortunistic fungal $athogen Aspergillus fumigates. Nature. 2 !; 4&,9 4,+B4. #ace J8. A 1enCoyl $ero@ide-sulfur crea/ for acne 7ulgaris. 5an .ed Assoc J. +'-&; '3"-%9 2&2-4. #earson =J, 3awson HJ, <acAey HB. Natural occurring le7els of di/ethyl sulfo@ide in selected fruits, 7egeta1les, grains and 1e7erages. J Agric 4ood 5he/. +'!+; 2'"&%9 + !'B'+. #rachayasittiAul S, Bura$aruangsang #, Jorachartcheewan A, IsaranAura-Na-Ayudhya 5, 0uchirawat S, #rachayasittiAul *, et al. Anti/icro1ial and antio@idant acti7ity of 1ioreacti7e constituents fro/ $ydnop+ytum formicarum. JacA .ol. 2 !; +39' 4B2+. 0adulo7ic NS, 3iAic .S, StoEano7ic-0adic FF. A new anti/icro1ial glucosinolate autolysis $roduct, 4isothiocyanato1utanoic acid, fro/ the diffuse Jallflower 4Erysimum diffusum59 .ethyl 4isothiocyanato1utanoate, a long unrecogniCed artifact of the isolation $rocedure^. 4ood che/. 2 ++; +2'9 +2&-3 . 0adulo7ic NS, 3iAic .S, StoEano7ic-0adic FF. Anti/icro1ial 7olatile glucosinolate autolysis $roducts fro/ $ornangia petraea "<.% 0ch1. "Brassicaceae%. #hytoche/ <ett. 2 +2; &9 3&+-,. 0ani I, AAhund S, A1ro H. Anti/icro1ial $otential of seed e@tract of 3ap+anus sativus. #aA J Bot. 2 !; 4 9 +,'3B!. 0asA <, Andreasson 8, 8A1o/ B, 8riAsson S, #onto$$idan B, .eiEer J, et al. .yrosinase9 >ene fa/ily e7olution and her1i7ore defense in Brassicaceae. #lant .ol Biol. 2 ; 429 '3++3. 0osa 8AS, 0odriges #.4. =owards a /ore sustaina1le agriculture syste/9 =he effect of glucosinolates of the control of soil1orne diseases. =he J Hort Sci Biotechnol. +'''; ,49 --,B,4. 0oy S#, NiranEan 5., Jyothi =., ShanArayya .., *ishawanath M., #ra1hu M, et al. Antiulcer and antiinfla//atory acti7ity of aerial $arts of Enicostemma littorale Blu/e. J ]oung #har/. 2 + ; 2"4%9 3-'-,3. Sahoo S, Mar 3., .oha$atra S, 0out S#, 3ash SM. Anti1acterial acti7ity of $y.ant+us enneaspermus against

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selected 6=I $athogens. Indian J #har/ Sci. 2 -; -!"&%9 -&3-&. -&3 Shin IS, .asuda H, Naohide M. Bactericidal ctericidal acti7ity of wasa1i "1asa.ia 2aponica% against $elico.acter pylori. Int J 4ood .icro1iol. 2 4; '49 2&&B B-+. Song, <, .orrison, JJ, Botting, N#, I =hornalley, #J. Analysis of glucosinolates, isothiocyanates, and a/ine degradation $roducts in 7egeta1le ta1le e@tracts and 1lood $las/a 1y <5B.S?.S. <5 .S?.S. Analytical Bioche/istry. 2 &; 34,, 234B243. Sousa 5, =a7eira ., *alentao #, 4ernandes 4, #ereira JA, 8ste7inho <. Inflorescences of Brassicaceae s$ecies as source of 1ioacti7e co/$ounds9 A co/$arati7e study. 4ood 5he/. 2 !; ++ 9 '&3--+. '&3 =ian \, 0osselot, 0A and SchwartC, SJ. \uantitati7e deter/ination of intact glucosinolates in 1roccoli, 1roccoli s$routs, Brussels s$routs, and cauliflower 1y high-$erfor/ance hig liGuid chro/atogra$hyBelectros$ray electros$ray ioniCationioniCation tande/ /ass s$ectro/etry. Analytical Bioche/istry, 2 &; 343, '3B''. '3 *alette <, 4ernandeC (, #olain S, <iCCani-5u7elier <iCCani 5u7elier <, <oiseau A. 5he/ical co/$osition of the 7olatile e@tracts fro/ Brassica oleraceae var,.otrytis N0o/anescoN cauliflower seeds. 4la7our 4rag J. 2 &; 2+9 + ,-+ , . *alette <, 4ernandeC (, #olain S, <oiseau A, <iCCani-5u7elier <iCCani 5u7elier <, <e7ieil 0, 0estier <, et al. *olatile constituents fro/ 0o/anesco cauliflower. 4ood 5he/. 2 3; ! 9 3&3-!. 3& *an <angenho7e HJ, 5ornelis 5#, Scha/$ N.. Identification of 7olatiles e/itted during the 1lanching $rocess of 1russels s$routs and cauliflower. J Sci 4ood Agric. +''+; &&"3%9 4!3B,. 4!3 *erAerA 0, Schreiner ., Mru/1ein A, 5isAa 8, Holst B, 0owland I, de SchriE7er 0, Hansen ., >erha_user 5, .ithen 0, 3eAAer ., et al. >lucosinolates in Brassica 7egeta1les9 the influence of the food su$$ly chain on intaAe, 1ioa7aila1ility and hu/an health. .ol Nut 4ood 0es. 2 '; &39 S2+'B-& S2+' *ig A#, 0a/$al >, =hind =S, Arora rora S. Bio-$rotecti7e Bio effects of glucosinolates B A re7iew. <J= - 4ood Sci =echnol. 2 '; 429 +&-+B,2. *olden J, Bengtsson >.B, Hansen ., Borge >IA, JicAlund =. >lucosinolates, <-ascor1ic < ascor1ic acid, total $henols, anthocyanins, antio@idant ca$acities and color colo in cauliflower "Brassica oleracea %. ssp. .otrytis%; effects of longter/ freeCer storage. 4ood 5he/. 2 '; ++29 '-,B,-. '-, Jall1anA B8, Jheatley >A. *olatile constituents fro/ cauliflower and other crucifers. #hytoche/. +',-; +&9 ,-3B-.

3igure &: =he general structure of glucosinolates and their enCy/atic degradation $roducts "ada$ted fro/ 0asA et al. "2 %.

,4

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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3igure ): H#<5?.S charts of Brassica oleraceae 7ar. 1otrytis "Soultany culti7ar%

,&

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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A' Glucosinolates: Basic structure9 G4S "`-thioglucoside-N- hydro@yl sulfate /oiety%

OH HO S R C N O OH CH2OH G4S OSO3
OH CH2 & GLS O S GLS GLS

#7

GLS ) O S . +

H2C CH2 GLS GLS GLS

S ,

GLS /

0 N H

O N

GLS

O S GLS N H && &) &2

OH OH OH OH HO O HO OH HO OH OH &, O OH O O O O OH OH OH OH OH OH OH OH O O OH OH O O O OH O OH

GLS

GLS

B'
HO HO OH &+ HO HO O

3ree 3lavonoid and5or glycosoids:

O OH O O O OH OH OH O O O

HO &HO

O OH

O HO OH OH O HO OH OH OH O O O O &. O O O

OH OH HO O O HO &/ HO HO O HO O O

OH OH O HO OH OH O OH OH O &0 HO O O HO O

O O

OH

HO OH OH

OH

C'

3ree 6"enolic acids and5or glycosides:

HO HO HO HO O O O OO O &1 OH HO HO OH O HO O HO OH O )& OH OH O OH OH O HO HO OH O O O OH O O O O O O OH O )2 O HO O HO HO OH OH O OH

O HO

,-

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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8' 6"ytoale9ins: Caulile9in C

NC )) N H3CO 3igure +: H#<5?.S identified constituents of Brassica oleraceae 7ar. 1otrytis "Soultany culti7ar%

:able &: *olatile co/$onents of Brassica oleraceae 7ar. 1otrytis "Soultany culti7ar%
5o/$onents 0I1 .Q "/?C% .ain frag/ents "/?C% <ea7es "D% Inflorescences "D% 4resh "D% .24 .24 .& .+! .!2 2.3 4roCen "D% .2+ tr tr .+2 3.!! +&.!! .&3 .++ +. + .2 .'' Ste/s "D%

3"8%-He@en-+-ol 4urfural #-@ylene 2"8%-He@anal +-$ro$yl-cyclo$roane )-@ylene 3"F%-He@en-+-ol Allylisothiocyanate Butane-+-isothiocyanate =risul$hide di/ethyl 2,4"8,8%-He$tadienal 2-#entyl furan 3"F%-He@ene-+-ol-acetate BenCene acetaldehyde ,"8%-.ethyl-+,--dio@as$iroX4.&Y decane +-".ethylthio%-3-$entanone )cta-3"8%,&"8%-dien-2-one --".ethylthio%he@-&-ene-3-ol 2-/ethyl-+H-indene +-o@o-+.2.4-trithiolane 3-.ethylthio-he@anal 3eca/ethyl-cyclo$entasilo@ane2 3-8thyl1enCaldehyde 2,-,--=ri/ethyl-+-cyclohe@ene-+acetaldehyde 4-.ethyl-3-"2-/ethyl$ro$-+-enyl%$yridine ,-/ethyl-2.3-dihydroisoindole-+-one 3odeca/ethyl- cyclohe@asilo@ane2

,-' ,'c !23 c c !3& !!, '+' '3& '4, ',3 ',' + 4 c + 4& + &, c c c c c c c c c c

+ '+ + !4 + + '' ++& +2++ +3! +42 +2 +&! +32 +24 +4+3 +4 +43, +34 +-+4, +4, 444

-,.2 -'.+ '+.+ &&.+ &-.2 '+.+ &&.2 ''.+ ++&. +2-. !+.+ !+.+ -,.2 '+.+ !4.+ +32. '&.+ ,3.+ !2.+ -+.+ , .2 3&&.2 ,&.+ !'.+ +4,.+ +4,.+ 34+.+

&-.2 '&.+ -,.2 -'.+ &&.2 + -.2 -,.2 '+.+ -+.+ ,'.+ &3.+ &,.2 !2.2 ,3.2 !,.+ &,.2 !+.+ !!.+ &4.2 +4 .+ &&.2 2-,.+ +34.+ +&+.2 ++-.+ ++-.+ 42'.2

&&.2 '4.+ &&.2 '&.+ -'.2 + &.2 !2.2 ,2.+ !&.+ +++. ++ .2 + &.+ !+.2 '2.+ '+.+ ,&.+ ,'.2 &,.2 +3 .+ ,'.+ +4-.+ ,3.+ +33.+ + ,.2 +32.+ +32.+ ,3.+

+!.!.,, +.3+ .+.+' .32 .+2 .+ -

.++ .3, .&' tr 24. tr .34 .+4 tr +.3, .+3 +.'&.,'

,,

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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:able &: (Contin')

5yclohe$tane 6nAnown "+% I/idaCole-4-car1o@a/ide "/ethylthio%-"chloro/ethylthio% /ethane Nonanal +-/ethyl-N-nitro-2"+H%-$yridini/ine =etradeca/ethyl Bcyclohe$tasilo@ane2 Butylated hydro@yl toluene 2,3,&-=rithiahe@ane 4-".ethylthio%1utyl cyanide 2-#henylethyl cyanide 3i/ethyl tetrasul$hide Beta-5yclocitral Indole Anethol "F% 4-.etho@y-BenCaldehyde Anethol "8% 3eca-2"8%,4"8%-dienal Anisyl acetone Beta-3a/ascone =etradecane 2-#henylethyl isothiocyanate =rans-1-ionone !,!-3i/ethyls$iro"4.-%undecane--,+ -dione &-iso$ro$enylo@y/ethylene-3,3-di/ethyl-cyclohe@anone He@adecane +-"iso$entylsulfinyl% $entane +.3.&.,-=etraethyl-+-ethyl1uto@ysilo@ycyclotetrasilo@ane2 3-/ethyl$yraColo1is"'-1ora1icycloX3.+.+Ynonane%o@ide >uaiol 2,3-dihydro-+,+,3-tri/ethyl-3-$henyl-+-H-Indene BH=-Guinone-/ethide )ctadecane 2.4-3i$henyl-4-/ethyl-+"8%-$entene 4-"3.4-di/etho@y1enCylidine%-+-"4-nitro$henyl%-3-$henyl-2$yraColin-&-one 2.4-3i$henyl-4-/ethyl-2"8%-$entene -.+ .+4-tri/ethyl-2-$entadecanone 6nAnown "2% 4-5u/yl-$henol 5is,cis,cis-,,+ ,+3-he@adecatrienal 4-t-1utyl-2-"di/ethyl1enCyl%$henol .ethyl $al/itate #hytol 3i1utyl iso$hthalate ester 3,4-dihydro-,,+2-di/ethyl1enCXaYanthracene 2-"+,+-di/ethylethyl%-4-/ethyl-+-$henylethyl-$henol 8icosa/ethyl-cyclodecasilo@ane2 6nAnown "3% 3ihydro-cis-al$ha-co$aene-!-ol He@adeca/ethyl-he$tasilo@ane &.,- 3iiso$ro$yl @anthone-2-car1o@ylic acid '.+ -3i/etho@y-2.3-dihydroanthracene-+.4-dione BisX4-diethyla/inoY$henyl /ethanone --aCa-&,,,+2,+4-tetrathia$entacene =etracosa/ethyl-cyclododecasilo@ane2 c e c c + ,& c c c + '4 ++42 ++!3 ++!, ++'4 +24! +24' +2&+ +2!2 +2!2 +2' +3'2 +4 +4+' +4-+ c c +c c c ++,+c +! c c c c f c c c +' 4 +'42 2 2& c c c g c c c c c c '! +++ +42 +42 +&3 &+' 22 +4 +2' +3+ +&! +&2 ++, +4! +3+4! +&2 +,! +'2 +'! +-3 +'2 2 ! +'4 22+' 442 34 222 23& 2+! 2&4 2342' 232-! 2+2 234 2-! 2, 2'2,! 2&! 2-! ,4+ 222 &33 324 2, 324 3&& !!' &&.2 +++.2 -+.+ '&.+ '+.+ 2!+.+ 2 &.3 -+.+ -+.+ '+.+ +&!. +3,.2 ++,.+ +4!.2 +3&.+ +4!.2 !+.+ +2+.+ +,,.2 &,.2 '+.+ +,,.2 !3.2 +-3.+ &,.2 , .2 3&&.2 ,3.2 2 +.2 22+.2 +-+.+ &,.2 ++'.2 42'.2 +43.2 &!.2 +',.2 ,'.+ +,&.2 ,4.+ ,+.2 +4'.+ +' .2 2&3.2 2!+.+ 2 +.2 3&&.2 3 '.3 2&3.3 3 '.3 22+.+ 3&&.2 '!.2 &,.2 '3.+ &,.2 ,&.+ ,3.+ 22 .3 +4 .+ ++&.+ +3+.+ ,'.+ +&2.2 ' .+ +4,.2 +3-.+ +4,.2 ++,.+ +-4.2 +'2.2 ,+.2 +-3.+ +23.2 &&.2 +3&.+ ,+.2 &&.2 ,3.+ 2!+.+ +-+.2 +43.2 2 3.2 ,+.2 '+.+ ,3.2 22+.2 ,+.2 2+2.2 -,.2 2&3.2 !,.+ &,.2 +& .+ 2&!.3 +4'.+ ,3.2 +-+.2 22+.+ 2&2.3 2-!.3 3+ .3 3&&.+ 2!+.+ !,.+ ,3.2 + ,.+ &&.2 +&3.+ 32,. 2 -.2 ,'.+ -2.+ ,'.+ -4. + '.2 !'.+ +2+.+ ,,.+ ++,.+ +2+.+ +22.+ +23.2 !&.2 + &.2 +3&.2 +-4.2 +'3.+ !&.2 +4-.2 3&-.2 22+.2 +4'.+ 222.2 2+-.2 !&.2 +2 .2 3&&.+ 23-.2 &,.2 &&.2 '+.+ 2-!.3 &&.2 +23.2 223.2 '&.2 2-!.3 +4,.+ !+.2 +4,.+ 324.3 2&4.2 324.3 2!+.+ ,3.2 3.'' .3! .++ 3.'3 tr .4+ .4+ .&3 +.2& tr .2.3, +.,+ .4& .4! tr +.&3 3. ! &.+ .+3 tr .-4 2.43 2.,, .+' .!+ .22 .2, .+3 tr 2. , -. +.2, .43 .2! 2.3! .2+ 2.,+ &.+! -.2+ .3' 4.'2 '.-' .-, .-.3 ..3! .+& tr -.&, .! .+! .23 ,. ' tr .+, -.,&.-3 .2& tr 2.,& .34 .34 .+tr .+' tr tr -. ! .+4 tr &.!+ tr tr tr .2! &.3+

+.4+ -.!! 3.!2

tr 4.+' 4.-&

,!

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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:able &: (Contin')

!1-/ethyl-2-$ro$yl-2,3,4,&,-,,,!,!1octahydro-+H-3a-ACa-as-indacene 2.3.4.&.--$entachloro-BenCa/ine Iron, /onocar1onyl-"+.3-1utadiene-+.4dicar1onic acid,diethyl ester%a,aa di$yridyl Heneicosane Stearic acid 3.3adichloro-1enCidene 3ocosane -+,3-di/ethyl-$yridoX3,2-dY$yri/idine2,4-"+H,3H%-dione BenCo"a% $henaCine &,+4-diol-8,8,F-+,3,+2-nonadecatriene '-"3-1utenyl%-anthracene +4-1eta-$regnane =ricosane BicycloX3.3.+Ynonan-+-ol =etracosane #arasiticol 3-"4-N,N3i/ethyla/ino$henyl%$ro$onoic acid2-"dietho@y$hos$hinyl%-ethyl ester 3ehydro a1ietic acid #entacosane +--al$ha-/ethyl$regnenolone 2-"di$henyl/ethyl%-+-$henyl---he$ten2-ol He@acosane +-al$ha-/ethyltestoterone -,!-dichloro-2-X4-chloro$henylY-4acetylGuinoline He$tacosane 2,4-di"tri/etho@ysilo@y%--,,-"/ethylene dio@y%-2H-+,4-1enCo@aCin-3-one-,-N-1iGuinoline 3-al$ha-aceto@y-+,-hydro@y androstane )ctacosane SGualene ++-/etho@y 1enC"a%anthracene-,,+2dione Nonacosane )-ter$henyl-+35 =riacontane Annotinine 2.4.--tris"+-/ethyl-+-$henyl ethyl%$henol =ra$eCifoli@anthone di/ethyl ether 3,-,!-tri-tert-1utyl-&/etho@y$henanthrene-+,4-dione .usco$yridine N-/ethyl-3i1enC"8,>% isoindole )ctadeca/ethyl-cyclononasilo@ane2 8thyl 2-X"+-hydro@y-2-iso$ro$yl-&/ethyl%cyclohe@ylYhe@anoate 2,-+ ,+4-tetra/ethyl-$entadecane 3,4-dihydro@y/andelic acid 4=.S Bis"2-ethylhe@yl%$hthalate =etradeca/ethyl-he@asilo@ane2 4-"BenCyla/ino%-+,3-di$henyl-&,-,,,!tetrahydro-2"+H%-Guinolinone +&-nonacosanone 3odeca/ethyl-$entasilo@ane2 S;ectra found (<) (dentified com;onents d Sul;"er containing com;onents (<) c c c 2+' 2 ' 43! 2 4.2 &&.2 3&&.2 +'+.2 -'.2 42'.2 &&.2 2--.3 ,3.2 +.43 .-4.-&.3 4.&-

2+ 22 c 22 c c c c c 23 c 24 c c

2'2!4 2&2 3+ +'+ 23 2'4 232 2!! 324 +4+ 33! 3 2 3&&

&,.2 &&.2 2&2.3 &,.2 +'+.2 23 .3 &&.2 ++'.2 '&.2 &,.2 ',.+ &,.2 2!,.2 22+.2

,+.2 &,.2 +!&.2 ,+.2 2 4.2 &,.2 '&.2 + -.2 &&.2 ,+.2 ++,.2 ,+.2 3 2.3 3&&.2

!&.2 !2.2 +!4.2 !&.2 + -.+ &&.2 -'.2 2+!.3 !+.2 !&.2 ++&.+ !&.2 2 '.2 ,3.2

+.'' 2. 3 .+& .2& 3.!, &. & -

.+, .!4 .4.4& .34 .!+ .&' !.+! -

.+3

.+2 .2! 4.!

tr &.4&

c 2& c c 2c c 2, c c c 2! c c 2' c 3 c c c c c c c c c c c c c c c

3 3&2 33 3&3-3+4 3'4 3! 4-3 2&334 3'4 4+ 2!! 4 ! 234 422 2,& 44! 4 4 23+ 23+ --, 2'! 2-! 4,2 3' 4&! 4 422 3!4

2!&.3 &,.2 3+&.3 2'3.3 &,.2 +3&.+ 22+.2 &,.2 34+.+ +4+.2 2 &.2 &,.2 -'.2 2 &.2 &,.2 23+.2 &,.2 ++,.2 433.4 3'+.3 3'+.3 23+.2 23+.2 3&&.+ 3&&.+ ,+.2 3&&.+ +4'.+ 3&&.+ 3'+.3 22&.3 22+.2

23'.2 ,+.2 3+-.3 3,+.4 ,+.2 +'+.2 2!+.+ ,+.2 42'.2 2&-.3 2,4.3 ,+.2 !+.2 2!!.3 ,+.2 23 .2 ,+.2 ++!.2 44!.4 4 -.3 4 -.3 ++,.+ ++,.+ ,3.2 22+.+ &,.2 ,3.+ +-,.+ 22+.+ 4 -.3 24+.3 3&&.+

!,.2 !&.2 33 .3 3,2.4 !&.2 2- .3 +4,.2 !&.2 ,3.+ &&.2 2+!.3 !&.2 '&.2 2+'.3 !&.2 ++'.2 !&.2 ++&.+ 434.4 3'2.3 3'2.3 23 .2 23 .2 42'.2 2!+.+ !&.2 2!+.+ 2,'.2 42'.2 3'2.3 &,.+ 2!+.+

.4+ &.+' ,.,4 ++.44.+, 2.!2 .2+.3! .2' .!2 .3+ .2! .!4 .+,

2.3, +.22 .&& +.4 +. 4 !.3+ +,. , .2+ 4.-+.43 1+'+/ ,+ (-'.&)

.3, .44 3. ! .+ 2. ' .34 &.23 .2& &.+2 .4+ 2.4, .34 .++ .3 11'&+ ,. -(),',/)

4.3.+4 =r .&3 1+'/) ,. &2 (++'.,)

.4! .&2 10'/1 ,1 , (&',,)

,'

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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:able &: (Contin')

Isothiocyanates "D% 5yanides "D% Alcohols "D% Aldehydes "D% Metones "D% #henols "D% 8sters "D% 8thers "D% 5ar1o@ylic acids "D% .iscellaneous "D%
a

4"3 .--% 4" .'2% !"+,.&,% & "&.'4% 2 "3. 2% + "3.'3% 2 "+.'4% +'
"34.--%

+ " .+'% 2"+.-+% +" .22% ,"+-.,' + "2.,,% &"+4.'! 2 "+. !% + " .+,% 22
"4'.-!%

&"4.4,% -"2.,+% !"++.&3% + "'.-'% 3"+ .33% + "&.+2% +,

"3 .!+%

3 " .!3% 2 " .2,% + " . 3% 3 " .4&% ! "+2.&!% + "&.-3% 3 "4.!-% 2 " .& % +3
"3&.2%

5o/$ounds are listed in order of their elution ti/e fro/ a 3B-& colu/n. 5o/$ositional 7alues less than .+D are denoted as traces "tr% 1 0I b retention indices as deter/ined on 3B-& colu/n. c Structure confir/ed 1y "Jilley,N; .ass 4inder and Jilley , Nist &; .ass 4inder% li1raries without deter/ination of their retention indices due to their high /olecular weights "c 4 %; d Identified co/$onents less than .&D in each organ are not re$orted. e 6nAnown "+%9 2 + ".Q%, +++ "+ %, &, "!2.2%, ,3 "!2%, '& "&3.4%, -' "44.4% f 6nAnown "2%9 3!, ".Q%, 243 "+ %, +!& ",,.3%, && ",,.2%, '& ",3.,%, -' "-,.3% g 6nAnown "3%9 3+- ".Q%, 23! "+ %, +,+ "'4.!%, +, "-!%, ++' "&3.3%, +&, "&+.'% 29 #ossi1le artifact due to instru/entation.

:able ): H#<5?.S identified co/$onents and conditions of detection of Brassica oleraceae "Soultany culti7ar%
<ist of co/$ounds e7aluated 1y 8SI 0t "/in% 8SI transition "/?C% .S+?.S2? ".S3% 43-? 3,2? "34+? 2&!% 423?2-+?2+&?++4 3&!?2&'?24+?2 ' 4 -? 3, ? "334? 2,2% 3,2? 33-? "22-?+!!% ,,+?- '? "42'?2!4% '33?,,+? "- '?42'?2!4% 42 ? 2!&? "+'!% 44,?4 +? "32,?3 '% ',,?!+&?- '?42'?2!4 44,?2&'? "23!?+-'?+3'% 4,,?2&'?+-!?+3' - '?42'?2&4?+&+ 4,,? 44-?4+,?3!3 ,23?4''?2&'?+'3 +!-?+-!? "-% 4 !? 3,2? "34+? 3++% 4 2? +'4?+,' 3&4?+,-?+-3 2, ?22&? "+'-% 3!!?2! ? "2&3?23,% '- ? '+3? "-3&? 3,'% =i/e window "/in% .&3B&. .&+B4. .&2B4. 2.+ B,. .&4B!. 2. B+-. 2. B+-. 2.+ B!. 2.+3B!. ,. B++. ,.4 B++. + . B +3. '. B+-. +3. B +,. +3.2 B +,. 3.3 B!. 3.- B!. ,. B++. ,.2 B++. .&&B4. .&&B4. .&-B4. 3well ti/e "s% .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ .+ 5one 7oltage "*% 34 44 3! 4 4 3& 3& 4! 4! 3& 3& 3& 44 3& 3& 3, 42 3& 4& 3& 3& 4+ 5ollision energy "e*% 32 24 22 2& 2 3 3 23 3 3 2& 2& 4 2& 2, 2 23 2& 23 2& 2' 0elati7e D "<?4?S%

>lucora$hanin ".% >lucoi1erin "-% Sinigrin "&% >lucoi1er7erin "&&% >lucona$in ")% Mae/$ferol-3-diglucoside-,-glucoside "&+% Mae/$ferol-3-triglucoside-,-glucoside "&,% >lucoerucin ",% Mae/$ferol -,-o-glucoside "&-% Mae/$ferol -3-o-"sina$oyl%-so$horoside,-o-glucoside "&.% >luco1rassicin "0% 4-/etho@y-gluco1rassicin "&2% 0utin "&/% Neogluco1rassicin "1% +-sina$oyl-2-feruloyldiglucoside "&1% 5aulile@in 5 "))% >lucotro$aeolin "/% n-He@yl?/ethyl$entyl glucosinolates "&)% 5hlorogenic acid ")&% A$igenin "&0% #rogoitrin "+% +,2,2N-=risina$oyldiglucoside ")2%

2 2.+ 2.3 2., 3.3 3.& 4.4 4.& 4., & &.+ &.! &.' ,.3 '.4 '.+-.2'.& 2'.32.' 3&.! 4!.-

=5=5tr =52',5)') tr5=5&') =5tr5+'& =5tr5+') )'15=5= )'/5=5= =5=5&') /',52'&5)') &')5=5= =5tr5&'+ )')5=5+') )'&5=5= .'/5=5/'+ &'+5=5= =52'-52'& =5tr5= )52'15+'0 +'&52'15+'& =5&',5= =5tr5tr =5tr5=

"8SI% B .S-scan /ode "ca$illary 7oltage B 2.& A*; cone 7oltage B 3 *%. 3aughtersN /ode "cone 7oltage B 3& *, collision energy B 3 e*%. "<?4?S%9 <ea7es?Inflorescence?Ste/s. tr9 traces; = 9 A1sent.

Journal of Biology, Agriculture and Healthcare ISSN 2224-32 ! "#a$er% ISSN 222&- '3( ")nline% *ol.3, No.+,, 2 +3

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:able +: Anti/icro1ial acti7ity of 7olatiles of inflorescences, lea7es and ste/s of Brassica oleraceae.
Sam;le :ested microorganisms 3ungi Aspergillus fumigatus "05.B 2&-4% !andida al.icans "05.B & 3&% Geotricum candidum "05.B & '-% /ric+op+yton mentagrop+ytes "05.B '2&% Gram=;ositive bacteria: )tap+ylococcus aureus "05.B + 2,% )tap+ylococcus epidermidis "05.B + 24% )treptococcus pyogenes "05.B + +&% Gram=negative bacteria: 0eisseria gonorr+oeae "05.B + ,-% Proteous vulgaris "05.B + !&% Kle.siella pneumoniae "05.B + '3% )+igella flexneri "05.B + &42% Pseudomonas aeruginosa "05.B + 43% Esc+eric+ia coli "05.B + &-% (nflorescences (fres"> fro?en) +!.4 V +.3' +2.+ V +.42 +3.3 V .'& +&.+V +.2+ +3.! V +.33 +&.3 V .'2 +-.3 V +.3' +&.'V .,' NA +,.- V .!' +-.3 V +.+, NA +!.& V .'2 4eaves Stems Standard Am;"otericin B 24.-V .+ 2+.!V .+2 2-.4V .2 2&.4V .+Am;icillin 2!.-V .+4 2&.2V .+! 2-.4V .34 Gentamycin +'.4V .+! 23.4V .3 2-.2V .+& 24.!V .24 +,.3V .+2 2&.3V .+!

NA '.3 V .&2 + .+ V .,& +2.!V .'+ NA NA NA NA NA +!.& V +.32 +-., V +.+& NA +,.2 V +.+,

+'., V +.2' +4.4 V +.-! +2.4 V +.3+ +!.2 V +.+' +-.- V .,+ +'.2 V +.2' +,.+ V +.+2 +&.& V +. + ++.'V .!3 +,.4 V +. ' +&.4 V +.2! NA +'., V +.3,

=he test was done using the agar diffusion techniGue, Jell dia/eter9 -. //, 05.B9 0egional 5enter for .ycology and Biotechnology Anti/icro1ial unit test organis/s. NA9 No acti7ity, data are e@$ressed in the for/ of /eanV S3.

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