Ibisobanuro
9,10-Anthraquinone (AQ) ni umwanda ufite ibyago bishobora gutera kanseri kandi biboneka mu cyayi ku isi. Umubare ntarengwa usigaye (MRL) wa AQ mu cyayi washyizweho n’ubumwe bw’ibihugu by’Uburayi (EU) ni 0,02 mg / kg. Inkomoko zishoboka za AQ mugutunganya icyayi nibyiciro byingenzi byabayeho byakozweho iperereza hashingiwe ku buryo bwahinduwe bwa AQ bwo gusesengura hamwe na gazi chromatografiya-tandem mass spectrometrie (GC-MS / MS). Ugereranije n’amashanyarazi nkisoko yubushyuhe mugutunganya icyayi kibisi, AQ yiyongereyeho inshuro 4.3 kugeza 23.9 mugutunganya icyayi hamwe namakara nkisoko yubushyuhe, irenga kure 0.02 mg / kg, mugihe urwego rwa AQ mubidukikije rwikubye gatatu. Ikintu kimwe cyagaragaye no gutunganya icyayi cya oolong munsi yubushyuhe bwamakara. Intambwe zifitanye isano itaziguye hagati yamababi yicyayi numwotsi, nko gutunganya no gukama, bifatwa nkintambwe nyamukuru yumusaruro wa AQ mugutunganya icyayi. Urwego rwa AQ rwiyongereye hamwe nigihe cyo guhura kwinshi, byerekana ko urugero rwinshi rwa AQ ihumanya icyayi rushobora guturuka kumyotsi iterwa namakara no gutwikwa. Ingero mirongo ine ziva mu mahugurwa atandukanye afite amashanyarazi cyangwa amakara nk’isesengura ry’ubushyuhe, yavuye kuri 50.0% −85.0% na 5.0% −35.0% yo gutahura no kurenza igipimo cya AQ. Byongeye kandi, AQ ntarengwa ya 0,064 mg / kg yagaragaye mu bicuruzwa byicyayi hamwe namakara nkisoko yubushyuhe, byerekana ko urugero rwinshi rwa AQ rwanduye mubicuruzwa byicyayi rushobora gutangwa namakara.
Ijambo ryibanze: 9,10-Anthraquinone, Gutunganya icyayi, Amakara, isoko yanduye
IRIBURIRO
Icyayi gikozwe mu bibabi by'icyatsi kibisi cyose cyitwa Camellia sinensis (L.) O. Kuntze, ni kimwe mu binyobwa bizwi cyane ku isi kubera uburyohe bushya kandi bifite akamaro ku buzima. Mu 2020 ku isi hose, umusaruro w'icyayi wariyongereye ugera kuri toni miliyoni 5.972, zikubye kabiri mu myaka 20 ishize [1]. Ukurikije uburyo butandukanye bwo gutunganya, hari ubwoko butandatu bwingenzi bwicyayi, harimo icyayi kibisi, icyayi cyirabura, icyayi cyijimye, icyayi cya oolong, icyayi cyera nicyayi cyumuhondo [2,3]. Kugirango hamenyekane ubuziranenge n'umutekano by'ibicuruzwa, ni ngombwa cyane gukurikirana urwego rwanduye no gusobanura inkomoko.

Kumenya inkomoko yanduye, nkibisigisigi byica udukoko, ibyuma biremereye nibindi byangiza nka hydrocarbone ya polycyclic aromatic hydrocarbone (PAHs), nintambwe yambere yo kurwanya umwanda. Gutera mu buryo butaziguye imiti y’ubukorikori mu gihingwa cy’icyayi, ndetse no gutwarwa n’umwuka uterwa n’ibikorwa hafi y’ubusitani bw’icyayi, ni byo soko nyamukuru y’ibisigazwa byica udukoko mu cyayi [4]. Ibyuma biremereye birashobora kwegeranya mu cyayi kandi biganisha ku burozi, bukomoka ahanini ku butaka, ifumbire ndetse n’ikirere [5-7]. Ku bijyanye n’indi myanda igaragara mu buryo butunguranye mu cyayi, byari bigoye kuyimenya bitewe nuburyo bugoye bwurwego rwicyayi rutanga umusaruro harimo guhinga, gutunganya, gupakira, kubika no gutwara. PAHs mu cyayi yavuye mu gushira imyuka y’ibinyabiziga no gutwika ibicanwa byakoreshejwe mu gutunganya amababi y’icyayi, nk'inkwi n’amakara [8-10].

Mu gihe cyo gutwika amakara n’ibiti, habaho umwanda nka okiside ya karubone [11]. Kubera iyo mpamvu, birashoboka cyane ko ibisigazwa by’ibi bihumanya byavuzwe haruguru biboneka mu bicuruzwa bitunganijwe, nk'ingano, ibigega byanyweye hamwe n’amafi y’injangwe, ku bushyuhe bwinshi, bikaba bibangamira ubuzima bw’abantu [12,13]. PAHs iterwa no gutwikwa ikomoka ku guhindagurika kwa PAHs ikubiye mu bicanwa ubwabyo, ubushyuhe bwo hejuru cyane bwangirika bw’ibintu bya aromatiya hamwe n’imikorere ivanze hagati ya radicals yubuntu [14]. Ubushyuhe bwo gutwika, igihe, hamwe na ogisijeni nibintu byingenzi bigira ingaruka kumihindagurikire ya PAHs. Hamwe n'ubwiyongere bw'ubushyuhe, ibirimo PAHs byabanje kwiyongera hanyuma bigabanuka, kandi agaciro k'impanuka kabaye kuri 800 ° C; Ibigize PAHs byagabanutse cyane kugirango bikurikirane igihe cyo gutwika igihe cyari munsi yurugero rwiswe 'igihe cyimbibi', hamwe no kwiyongera kwa ogisijeni mu kirere cyaka, imyuka ya PAHs yagabanutse cyane, ariko okiside ituzuye byabyara OPAH nizindi nkomoko [15 −17].

9,10-Anthraquinone (AQ, CAS: 84-65-1, Igishushanyo 1), ogisijeni irimo ogisijeni ikomoka kuri PAHs [18], igizwe ninzinguzingo eshatu zegeranye. Yashyizwe ku rutonde nka kanseri ishobora gutera (Itsinda 2B) n'ikigo mpuzamahanga gishinzwe ubushakashatsi kuri kanseri mu 2014 [19]. AQ irashobora kwangiza uburozi bwa topoisomerase II kandi ikabuza hydrolysis ya adenosine triphosphate (ATP) na ADN topoisomerase II, bigatuma ADN imeneka kabiri, bivuze ko kumara igihe kirekire mubidukikije birimo AQ no guhura kurwego rwo hejuru rwa AQ irashobora gutuma ADN yangirika, ihinduka kandi ikongera ibyago byo kurwara kanseri [20]. Nkingaruka mbi ku buzima bwabantu, AQ ntarengwa y’ibisigisigi (MRL) ya 0,02 mg / kg yashyizwe mu cyayi n’umuryango w’ubumwe bw’ibihugu by’Uburayi. Dukurikije ubushakashatsi bwatubanjirije, kubitsa AQ byatanzwe nkisoko nyamukuru mugihe cyo guhinga icyayi [21]. Na none kandi, hashingiwe ku ngaruka z’ubushakashatsi bwakozwe muri Indoneziya icyatsi kibisi n’umukara, biragaragara ko urwego rwa AQ rwahindutse ku buryo bugaragara kandi umwotsi w’ibikoresho bitunganyirizwa watanzwe nkimwe mu mpamvu zingenzi [22]. Nyamara, inkomoko nyayo ya AQ mugutunganya icyayi yakomeje kuba ingorabahizi, nubwo hari ibitekerezo bimwe na bimwe byerekana inzira ya chimique ya AQ [23,24], byerekana ko ari ngombwa cyane kumenya ibintu byingenzi bigira ingaruka kurwego rwa AQ mugutunganya icyayi.

Igishushanyo 1. Imiti yimiti ya AQ.

Bitewe n'ubushakashatsi bwakozwe ku ishyirwaho rya AQ mu gihe cyo gutwika amakara ndetse n’akaga gashobora guterwa n’ibicanwa mu gutunganya icyayi, hakozwe ubushakashatsi bugereranya bwo gusobanura ingaruka zituruka ku masoko y’ubushyuhe kuri AQ mu cyayi no mu kirere, isesengura ryinshi ku ihinduka ry’ibirimo AQ ku ntambwe zitandukanye zo gutunganya, zifasha kwemeza inkomoko nyayo, imiterere yibibaho hamwe n urugero rwumwanda wa AQ mugutunganya icyayi.

IBISUBIZO
Uburyo bwo kwemeza
Ugereranije nubushakashatsi bwatubanjirije [21], uburyo bwo kuvoma amazi-bwamazi bwahujwe mbere yo gutera inshinge GC-MS / MS hagamijwe kunoza ibyiyumvo no gukomeza amagambo yibikoresho. Igishushanyo cya 2b, uburyo bunoze bwerekanye iterambere ryinshi mugusukura icyitegererezo, umusemburo wabaye mwiza mubara. Mu gishushanyo cya 2a, icyerekezo cyuzuye cya scan (50−350 m / z) cyerekanaga ko nyuma yo kwezwa, umurongo wibanze wumurongo wa MS wagabanutse kuburyo bugaragara kandi impinga nke za chromatografique zari zihari, byerekana ko umubare munini wibintu bivangavanze byakuweho nyuma ya kuvoma amazi.

Igishushanyo 2. (A) Gusikana byuzuye byerekana urugero mbere na nyuma yo kwezwa. (b) Ingaruka yo kweza uburyo bunoze.
Kwemeza uburyo, harimo umurongo, kugarura, imipaka yo kubara (LOQ) n'ingaruka za matrix (ME), bigaragara mu mbonerahamwe ya 1. Birashimishije kubona umurongo hamwe na coefficente yo kwiyemeza (r2) hejuru ya 0.998, kuva kuri 0.005 kugeza 0.2 mg / kg muri matrix yicyayi hamwe na acetonitrile solvent, no mubyitegererezo byikirere bifite intera ya 0.5 kugeza 8 μg / m3.

Isubiranamo rya AQ ryasuzumiwemo ibintu bitatu byibanze hagati yibipimo byapimwe kandi bifatika mu cyayi cyumye (0.005, 0.02, 0.05 mg / kg), icyayi gishya (0.005, 0.01, 0.02 mg / kg) hamwe nicyitegererezo cyikirere (0.5, 1.5, 3 μg / m3). Kugarura AQ mu cyayi kuva kuri 77,78% kugeza kuri 113.02% mu cyayi cyumye naho kuva kuri 96.52% kugeza kuri 125.69% mu cyayi, naho RSD% iri munsi ya 15%. Kugarura AQ mu byitegererezo by'ikirere byari hagati ya 78.47% na 117.06% hamwe na RSD% munsi ya 20%. Ikigereranyo cyo hasi cyane cyamenyekanye nka LOQ, ni 0.005 mg / kg, 0.005 mg / kg na 0.5 μg / m³ mu cyayi, icyayi cyumye hamwe n’icyitegererezo cy’ikirere. Nkuko bigaragara mu mbonerahamwe ya 1, matrix yicyayi cyumye nicyayi cyicyayi byongereye gato igisubizo cya AQ, biganisha kuri ME ya 109.0% na 110.9%. Kubijyanye na matrix yicyitegererezo cyindege, ME yari 196.1%.

Urwego rwa AQ mugihe cyo gutunganya icyayi kibisi
Mu ntumbero yo kumenya ingaruka zituruka ku bushyuhe butandukanye ku cyayi no gutunganya ibidukikije, icyiciro cy’amababi mashya cyagabanyijwemo amatsinda abiri yihariye kandi gitunganyirizwa ukundi mu mahugurwa abiri yo gutunganya mu kigo kimwe. Itsinda rimwe ryahawe amashanyarazi, irindi ryahawe amakara.

Nkuko bigaragara ku gishushanyo cya 3, urwego rwa AQ rufite amashanyarazi nkuko isoko yubushyuhe yari hagati ya 0.008 na 0.013 mg / kg. Mugihe cyo gutunganya, gutobora amababi yicyayi biterwa no gutunganyirizwa mu nkono ifite ubushyuhe bwinshi byatumye AQ yiyongera 9.5%. Hanyuma, urwego rwa AQ rwagumye mugihe cyo kuzunguruka nubwo gutakaza umutobe, byerekana ko inzira zumubiri zidashobora kugira ingaruka kurwego rwa AQ mugutunganya icyayi. Nyuma yintambwe yambere yo kumisha, urwego rwa AQ rwiyongereyeho gato kuva 0.010 rugera kuri 0.012 mg / kg, hanyuma rukomeza kuzamuka rugera kuri 0.013 mg / kg kugeza rwongeye gukama. PFs yerekanaga cyane itandukaniro muri buri ntambwe, yari 1.10, 1.03, 1.24, 1.08 mugukosora, kuzunguruka, kubanza gukama no kongera gukama. Ibisubizo bya PF byagaragaje ko gutunganya ingufu z'amashanyarazi byagize ingaruka nke kurwego rwa AQ mu cyayi.

Igishushanyo 3. Urwego rwa AQ mugihe cyo gutunganya icyayi kibisi hamwe namashanyarazi namakara nkisoko yubushyuhe.
Ku bijyanye n’amakara nkisoko yubushyuhe, ibirimo AQ byiyongereye cyane mugihe cyo gutunganya icyayi, biva kuri 0.008 bigera kuri 0.038 mg / kg. 338.9% AQ yariyongereye muburyo bwo gukosora, igera kuri 0.037 mg / kg, yarenze kure MRL ya 0,02 mg / kg yashyizweho n’umuryango w’ubumwe bw’ibihugu by’Uburayi. Mugihe cyo kuzunguruka, urwego rwa AQ rwiyongereyeho 5.8% nubwo ari kure yimashini ikosora. Mubanza gukama no kongera gukama, ibirimo AQ byiyongereyeho bike cyangwa byagabanutseho gato. PF ikoresha amakara nkisoko yubushyuhe mugukosora, kuzunguruka mbere yumisha no kongera gukama byari 4.39, 1.05, 0.93, na 1.05.

Kugirango hamenyekane isano iri hagati yo gutwika amakara n’umwanda wa AQ, ibibazo byahagaritswe (PMs) mu kirere mu mahugurwa munsi y’amasoko yombi byakusanyirijwe hamwe kugira ngo hasuzumwe ikirere, nk'uko bigaragara ku gishushanyo cya 4. Urwego rwa AQ rwa PM hamwe n’amakara nk inkomoko yubushyuhe yari 2,98 μg / m3, yikubye inshuro zirenga eshatu ugereranije n’amashanyarazi 0,91 μg / m3.

Igicapo 4. Urwego rwa AQ mubidukikije hamwe namashanyarazi namakara nkisoko yubushyuhe. * Yerekana itandukaniro rikomeye murwego rwa AQ murugero (p <0.05).

Urwego rwa AQ mugihe cyo gutunganya icyayi cya oolong Icyayi cya Oolong, gikorerwa cyane cyane muri Fujian na Tayiwani, ni ubwoko bwicyayi gisembuye igice. Kugirango urusheho kumenya intambwe nyamukuru yo kongera urwego rwa AQ n'ingaruka za lisansi zitandukanye, icyiciro kimwe cyamababi mashya cyakozwe mucyayi cya oolong hamwe namakara hamwe na gaze gasanzwe-amashanyarazi nkisoko yubushyuhe, icyarimwe. Urwego rwa AQ mu gutunganya icyayi cya oolong ukoresheje amasoko atandukanye y’ubushyuhe rwerekanwe ku gishushanyo cya 5. Ku gutunganya icyayi cya oolong hamwe n’imvange ya gaze isanzwe y’amashanyarazi, icyerekezo cya AQ cyari gihagaze munsi ya 0.005 mg / kg, cyari kimeze nkicyayi kibisi n'amashanyarazi.

Igicapo 5. Urwego rwa AQ mugihe cyo gutunganya icyayi cya oolong hamwe na gaze gasanzwe-amashanyarazi hamwe namakara nkisoko yubushyuhe.

Hamwe namakara nkisoko yubushyuhe, urwego rwa AQ mubyiciro bibiri byambere, byumye kandi bigakora icyatsi, mubyukuri byari bimwe nkibisanzwe bivangwa na gaze-amashanyarazi. Nyamara, inzira yakurikiyeho kugeza gukosorwa yerekanaga icyuho cyagutse buhoro buhoro, icyo gihe urwego rwa AQ rwavuye kuri 0.004 rugera kuri 0.023 mg / kg. Urwego rwapakiye intambwe yagabanutse rugera kuri 0.018 mg / kg, ibyo bikaba bishobora guterwa no gutakaza umutobe wicyayi utwara bimwe mubihumanya AQ. Nyuma yo kuzunguruka, urwego murwego rwo kumisha rwiyongereye kugera kuri 0.027 mg / kg. Mu gukama, gukora icyatsi, gutunganya, gupakira no gukama, PF yari 2.81, 1.32, 5.66, 0.78, na 1.50.

Kugaragara kwa AQ mubicuruzwa byicyayi hamwe nubushyuhe butandukanye

Kugirango hamenyekane ingaruka kuri AQ yibirimo byicyayi hamwe nubushyuhe butandukanye, icyitegererezo cyicyayi 40 kiva mumahugurwa yicyayi hifashishijwe amashanyarazi cyangwa amakara nkamasoko yubushyuhe, nkuko bigaragara mu mbonerahamwe ya 2. Ugereranije no gukoresha amashanyarazi nkisoko yubushyuhe, amakara yari afite menshi ibipimo by'iperereza (85.0%) hamwe na AQ ntarengwa ya 0.064 mg / kg, byerekana ko byari byoroshye gutera umwanda AQ umwotsi ukomoka ku gutwika amakara, kandi igipimo cya 35.0% cyagaragaye mu ngero z'amakara. Ikigaragara cyane ni uko amashanyarazi yari afite igipimo cyo hasi cyane cy’iperereza no hejuru ya 56.4% na 7.7%, hamwe na 0.020 mg / kg.

IKIGANIRO

Hashingiwe kuri PF mugihe cyo gutunganya hamwe nubwoko bubiri buturuka ku bushyuhe, byaragaragaye ko gukosora ari intambwe nyamukuru yatumye kwiyongera kwa AQ mu musaruro w’icyayi hamwe n’amakara no gutunganya ingufu z’amashanyarazi byagize ingaruka nke ku bikubiye muri AQ mu cyayi. Mugihe cyo gutunganya icyayi kibisi, gutwika amakara byatumaga imyotsi myinshi mugikorwa cyo gutunganya ugereranije nuburyo bwo gushyushya amashanyarazi, byerekana ko ahari imyotsi niyo soko nyamukuru yanduza AQ ituruka kumasoko yicyayi ako kanya mugutunganya icyayi, bisa nuburyo bwo kwerekana muri ingero za barbecue zanyweye [25]. Ubwiyongere bworoheje bwibintu bya AQ mugihe cyo kuzunguruka byerekanaga ko imyotsi iterwa no gutwikwa kwamakara itagize ingaruka gusa kurwego rwa AQ mugihe cyo gukosora, ahubwo no mubidukikije bitunganyirizwa kubera ikirere. Amakara nayo yakoreshejwe nkisoko yubushyuhe mugukama kwambere no kongera gukama, ariko muri izi ntambwe zombi ibirimo AQ byiyongereyeho gato cyangwa byagabanutseho gato. Ibi birashobora gusobanurwa nuko icyuma cyumuyaga gishyushye cyabujije icyayi kure yumwotsi uterwa no gutwika amakara [26]. Kugirango hamenyekane inkomoko yanduye, urwego rwa AQ mu kirere rwasesenguwe, bituma habaho itandukaniro rikomeye hagati yaya mahugurwa yombi. Impamvu nyamukuru yabyo nuko amakara akoreshwa mugukosora, kubanza gukama no kongera gukama byabyara AQ mugihe cyo gutwika kutuzuye. Izi AQ noneho zamamajwe mu tuntu duto duto duto nyuma yo gutwikwa n’amakara hanyuma zikwirakwizwa mu kirere, bizamura urugero rw’umwanda wa AQ mu mahugurwa [15]. Igihe kirenze, bitewe nubuso bunini bwubuso hamwe nubushobozi bwa adsorption yicyayi, utwo duce noneho twatuye hejuru yamababi yicyayi, bigatuma AQ yiyongera mubikorwa. Gutyo rero, gutwika amakara niyo nzira nyamukuru iganisha ku kwanduza AQ gukabije mu gutunganya icyayi, imyotsi ikaba intandaro y’umwanda.

Kubijyanye no gutunganya icyayi cya oolong, AQ yariyongereye mugutunganya hamwe nubushyuhe bwombi, ariko itandukaniro riri hagati yubushyuhe bwombi ryari rifite akamaro. Ibisubizo byanagaragaje ko amakara nk’isoko ry’ubushyuhe yagize uruhare runini mu kongera urwego rwa AQ, kandi gukosorwa byafashwe nkintambwe nyamukuru yo kongera kwanduza AQ mu gutunganya icyayi cya oolong gishingiye kuri PFs. Mugihe cyo gutunganya icyayi cya oolong hamwe na gaze isanzwe ya gazi-amashanyarazi nkisoko yubushyuhe, icyerekezo cyurwego rwa AQ cyari gihagaze munsi ya 0.005 mg / kg, ibyo bikaba byari bimeze nkicyayi kibisi gifite amashanyarazi, byerekana ko ingufu zisukuye, nkamashanyarazi nibisanzwe gaze, irashobora kugabanya ibyago byo kubyara AQ ibyanduye bitunganijwe.

Ku bijyanye n'ibizamini by'icyitegererezo, ibisubizo byagaragaje ko ikibazo cyo kwanduza AQ cyarushijeho kuba bibi iyo ukoresheje amakara nk'isoko ry'ubushyuhe aho gukoresha amashanyarazi, ibyo bikaba bishobora guterwa n'umwotsi uva mu gutwika amakara uhura n'amababi y'icyayi kandi ugatinda ku kazi. Nyamara, nubwo byagaragaye ko amashanyarazi ari isoko yubushyuhe isukuye mugihe cyo gutunganya icyayi, haracyari umwanda wa AQ mubicuruzwa byicyayi ukoresheje amashanyarazi nkisoko yubushyuhe. Ibintu bisa nkaho bisa nkibikorwa byasohotse mbere aho igitekerezo cya 2- alkenal hamwe na hydroquinone na benzoquinone cyatanzwe nkinzira ishobora guterwa imiti [23], impamvu yabyo izakorwaho ubushakashatsi mubushakashatsi buzaza.

UMWANZURO

Muri iki gikorwa, inkomoko zishobora kwanduza AQ mu cyayi kibisi na oolong byemejwe n’ubushakashatsi bugereranije bushingiye ku buryo bunoze bwo gusesengura GC-MS / MS. Ibyavuye mu bushakashatsi byashigikiye mu buryo butaziguye ko isoko nyamukuru ihumanya y’urwego rwo hejuru rwa AQ yari umwotsi watewe no gutwikwa, ibyo bikaba bitagize ingaruka ku cyiciro cyo gutunganya gusa ahubwo byanagize ingaruka ku mahugurwa. Bitandukanye no kuzunguruka no gukama, aho impinduka zo murwego rwa AQ zitagaragara, ibyiciro birimo uruhare rutaziguye rwamakara ninkwi, nko gukosora, ninzira nyamukuru aho AQ yanduye bitewe nubunini bwihuza hagati yicyayi n'umwotsi muri ibi byiciro. Kubwibyo, ibicanwa bisukuye nka gaze gasanzwe n amashanyarazi byasabwe nkisoko yubushyuhe mugutunganya icyayi. Byongeye kandi, ibyavuye mu bushakashatsi byerekanye kandi ko mu gihe hatabayeho umwotsi uturuka ku gutwikwa, hakiri izindi mpamvu zagize uruhare mu gushakisha AQ mu gihe cyo gutunganya icyayi, mu gihe umubare muto wa AQ nawo wagaragaye mu mahugurwa hamwe n’ibicanwa bisukuye, bigomba kurushaho gukorwaho iperereza mubushakashatsi buzaza.

IMIKORESHEREZE N'UBURYO

Reagents, imiti nibikoresho

Igipimo cya Anthraquinone (99.0%) cyaguzwe na Dr. Ehrenstorfer GmbH Company (Augsburg, Ubudage). D8-Anthraquinone yimbere (98,6%) yaguzwe muri C / D / N Isotopes (Quebec, Kanada). Anhydrous sodium sulfate (Na2SO4) na sulfate ya magnesium (MgSO4) (Shanghai, Ubushinwa). Florisil yatanzwe na Wenzhou Organic Chemical Company (Wenzhou, Ubushinwa). Impapuro z'ikirahure cya Mircro (90 mm) zaguzwe muri sosiyete ya Ahlstrom-munksjö (Helsinki, Finlande).

Icyitegererezo

Icyitegererezo cyicyayi kibisi cyatunganijwe no gukosorwa, kuzunguruka, kubanza gukama no kongera gukama (ukoresheje ibikoresho bifunze), mugihe icyayi cya oolong cyatunganijwe no gukama, gukora icyatsi (kunyeganyega no guhagarara amababi mashya ukundi), gukosora, kuzinga bipfunyitse, na kumisha. Ingero kuri buri ntambwe zegeranijwe inshuro eshatu kuri 100g nyuma yo kuvanga neza. Ingero zose zabitswe kuri −20 ° C kugirango zisesengurwe.

Ingero zo mu kirere zegeranijwe n'impapuro z'ibirahure (mm 90) hakoreshejwe urugero ruciriritse (PTS-100, Isosiyete ikora ibikoresho bya elegitoroniki ya Qingdao Laoshan, Qingdao, mu Bushinwa) [27], ikora 100 L / min kuri 4 h.

Ingero zikomeye zashyizwemo AQ kuri 0.005 mg / kg, 0.010 mg / kg, 0,020 mg / kg ku cyayi gishya, kuri 0.005 mg / kg, 0.020 mg / kg, 0.050 mg / kg ku cyayi cyumye no kuri 0.012 mg / kg . Nyuma yo kunyeganyega neza, ibyitegererezo byose byasigaye kuri 12 h, bikurikirwa no gukuramo intambwe zo gusukura.

Ibirungo byabonetse mu gufata g 20 yicyitegererezo nyuma yo kuvanga buri ntambwe, gushyushya kuri 105 ° C kuri 1 h, hanyuma gupima no gusubiramo inshuro eshatu no gufata agaciro kagereranijwe no kugabana nuburemere mbere yo gushyushya.

Icyitegererezo cyo gukuramo no gusukura

Icyitegererezo cy'icyayi: Gukuramo no kweza AQ mu cyitegererezo cy'icyayi byakozwe hashingiwe ku buryo bwatangajwe na Wang n'abandi. hamwe n'imihindagurikire y'ikirere [21]. Muri make, 1.5 g yicyitegererezo cyicyayi cyabanje kuvangwa na 30 μL D8-AQ (2 mg / kg) hanyuma bigasigara bihagarara kuminota 30, hanyuma bivangwa neza namazi 1.5 mL ya deionion hanyuma bigasigara bihagarara 30 min. 15 mL 20% acetone muri n-hexane yongewemo icyitegererezo cyicyayi hanyuma ihabwa min 15. Noneho ingero zashizwe hamwe na 1.0 g MgSO4 kuri 30 s, hanyuma zishiramo iminota 5, saa 11,000 rpm. Nyuma yo kwimurirwa muri flasque 100 mL, ama mL 10 yicyiciro cyo hejuru kama yahindutse umwuka hafi yumye kuri 37 ° C. 5 mL 2,5% acetone muri n-hexane yongeye gushonga ibivuye mumashanyarazi ameze nk'isaro kugirango bisukure. Inkingi yikirahure (cm 10 × 0.8 cm) yari igizwe kuva hasi kugeza hejuru yubwoya bwikirahure na 2g florisil, yari hagati yibice bibiri bya cm 2 Na2SO4. Noneho 5 mL ya 2,5% acetone muri n-hexane yahinduye inkingi. Nyuma yo gupakira igisubizo cyongeye gukemuka, AQ yavuzwe inshuro eshatu hamwe na mL 5, mL 10, mL 10 ya acetone 2,5% muri n-hexane. Intore zahujwe zimuriwe mumashanyarazi ameze nk'amapera hanyuma zihumuka hafi yumye munsi ya vacu kuri 37 ° C. Ibisigara byumye noneho byongeye gushyirwaho hamwe na mL 1 ya 2,5% ya acetone muri hexane ikurikirwa no kuyungurura binyuze kuri 0.22 µm yubunini bwa pore. Noneho igisubizo cyasubiwemo kivanze na acetonitrile ku kigereranyo cya 1: 1. Ukurikije intambwe yo kunyeganyega, subnatant yakoreshejwe mu gusesengura GC-MS / MS.

Icyitegererezo cy'ikirere: Kimwe cya kabiri cy'impapuro za fibre, gitonyanga hamwe na 18 μL d8-AQ (2 mg / kg), cyinjijwe muri mL 15 ya 20% ya acetone muri n-hexane, hanyuma ikabyara iminota 15. Icyiciro kama cyatandukanijwe na centrifugation saa 11,000 rpm kuminota 5 hanyuma igorofa yose yo hejuru ikurwaho mumashanyarazi ameze. Ibyiciro byose kama byavumbuwe hafi yumye munsi ya 37 ° C. 5 mL ya acetone 2,5% muri hexane yongeye gukuramo ibivuyemo kugirango bisukure kimwe nicyitegererezo cyicyayi.

Isesengura rya GC-MS / MS

Varian 450 ya chromatograf ifite ibikoresho bya Varian 300 tandem mass detector (Varian, Walnut Creek, CA, USA) yakoreshejwe mugukora isesengura rya AQ hamwe na software ya MS WorkStation 6.9.3. Ibintu Varian Ibintu bine capillary inkingi VF-5ms (30 m × 0,25 mm × 0,25 μ mm) yakoreshejwe mugutandukanya chromatografique. Gazi itwara, helium (> 99,999%), yashyizwe ku gipimo gihoraho cya 1.0 mL / min hamwe na gaze yo kugongana ya Argon (> 99,999%). Ubushyuhe bw'itanura bwatangiriye kuri 80 ° C kandi bufata iminota 1; yiyongereye kuri 15 ° C / min kugeza kuri 240 ° C, hanyuma igera kuri 260 ° C kuri 20 ° C / min ikomeza kuri 5min. Ubushyuhe bw'inkomoko ya ion bwari 210 ° C, kimwe n'ubushyuhe bwo kohereza umurongo wa 280 ° C. Ingano yo gutera inshinge yari 1.0 μL. Imiterere ya MRM igaragara mu mbonerahamwe ya 3.

Agilent 8890 gas chromatograf ifite ibikoresho bya Agilent 7000D triple quadrupole mass spectrometer (Agilent, Stevens Creek, CA, USA) yakoreshejwe mu gusesengura ingaruka zo kwezwa hamwe na software ya MassHunter 10.1. Agilent J&W HP-5ms GC Inkingi (30 m × 0,25 mm × 0,25 μ m) yakoreshejwe mugutandukanya chromatografique. Gazi itwara, Helium (> 99,999%), yashyizwe ku gipimo gihoraho cya 2,25 mL / min hamwe na gaze yo kugongana ya Azote (> 99,999%). Ubushyuhe bw'isoko rya EI bwahinduwe kuri 280 ° C, kimwe n'ubushyuhe bwo kwimura. Ubushyuhe bw'itanura bwatangiriye kuri 80 ° C kandi bumara iminota 5; yazamutseho 15 ° C / min kugeza kuri 240 ° C, hanyuma igera kuri 280 ° C kuri 25 ° C / min ikomeza iminota 5. Imiterere ya MRM igaragara mu mbonerahamwe ya 3.

Isesengura mibare
Ibiri muri AQ mumababi mashya byakosowe kubintu byumye bigabanywa nubushuhe kugirango ugereranye kandi usesengure urwego rwa AQ mugihe cyo gutunganya.

Impinduka za AQ mubyitegererezo byicyayi byasuzumwe hamwe na software ya Microsoft Excel hamwe na IBM SPSS Ibarurishamibare 20.

Ikintu cyo gutunganya cyakoreshejwe mugusobanura impinduka muri AQ mugihe cyo gutunganya icyayi. PF = Rl / Rf, aho Rf ni urwego rwa AQ mbere yintambwe yo gutunganya naho Rl ni urwego rwa AQ nyuma yintambwe yo gutunganya. PF yerekana kugabanuka (PF <1) cyangwa kwiyongera (PF> 1) mubisigisigi bya AQ mugihe cyintambwe yihariye yo gutunganya.

NJYE yerekana kugabanuka (ME <1) cyangwa kwiyongera (ME> 1) hasubijwe ibikoresho byisesengura, bishingiye ku kigereranyo cyimisozi ya kalibrasi muri matrix na solvent kuburyo bukurikira:

NJYE = (slopematrix / slopesolvent - 1) × 100%

Aho slopematrix ari umusozi wa kalibrasi ya curve muri matrix-ihuye na solvent, slopesolvent ni umusozi wa kalibrasi yo kugabanuka.

ICYEMEZO
Uyu murimo watewe inkunga n’umushinga w’ubumenyi n’ikoranabuhanga mu Ntara ya Zhejiang (2015C12001) na Fondasiyo y’ubumenyi y’Ubushinwa (42007354).
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