Abstract
9,10-Anthraquinone (AQ) wani gurɓataccen abu ne tare da yiwuwar ciwon daji kuma yana faruwa a cikin shayi a dukan duniya. Matsakaicin iyakar ragowar (MRL) na AQ a cikin shayi da Tarayyar Turai (EU) ta saita shine 0.02 mg/kg. An bincika yuwuwar tushen AQ a cikin sarrafa shayi da kuma manyan matakan faruwar sa bisa ga gyare-gyaren hanyar nazari na AQ da bincike na chromatography-tandem mass spectrometry (GC-MS/MS). Idan aka kwatanta da wutar lantarki a matsayin tushen zafi a sarrafa koren shayi, AQ ya karu da sau 4.3 zuwa 23.9 a sarrafa shayi tare da kwal a matsayin tushen zafi, wanda ya wuce 0.02 mg/kg, yayin da matakin AQ a cikin muhalli ya ninka sau uku. An lura da irin wannan yanayin a sarrafa shayin oolong a ƙarƙashin zafi na kwal. Matakan da ke da alaƙa kai tsaye tsakanin ganyen shayi da hayaƙi, kamar gyarawa da bushewa, ana ɗaukar su azaman manyan matakan samar da AQ a sarrafa shayi. Matakan AQ sun karu tare da tashin lokacin tuntuɓar, yana nuna cewa babban matakan gurɓataccen AQ a cikin shayi na iya samuwa daga tururin da gawayi da konewa ke haifarwa. Samfura guda arba'in daga tarurrukan bita daban-daban tare da wutar lantarki ko kwal kamar yadda aka bincika tushen zafi, sun fito daga 50.0% -85.0% da 5.0% -35.0% don ganowa da wuce ƙimar AQ. Bugu da ƙari, an lura da matsakaicin adadin AQ na 0.064 mg / kg a cikin samfurin shayi tare da kwal a matsayin tushen zafi, yana nuna cewa yawan ƙwayar AQ a cikin kayan shayi yana iya taimakawa ta hanyar kwal.
Mahimman kalmomi: 9,10-Anthraquinone, sarrafa shayi, Coal, Tushen gurɓatawa
GABATARWA
Tea da aka kera daga ganyen ciyawar da ba a taɓa gani ba ta Camellia sinensis (L.) O. Kuntze, yana ɗaya daga cikin shahararrun abubuwan sha a duniya saboda ɗanɗanonsa da fa'idodin kiwon lafiya. A cikin 2020 a duniya, samar da shayi ya karu zuwa metric ton miliyan 5,972, wanda ya ninka a cikin shekaru 20 da suka gabata[1]. Dangane da hanyoyin sarrafawa daban-daban, akwai manyan nau'ikan shayi guda shida, waɗanda suka haɗa da koren shayi, baƙar fata, shayi mai duhu, shayin oolong, farin shayi da shayin rawaya[2,3]. Don tabbatar da inganci da amincin samfuran, yana da matukar mahimmanci don saka idanu matakan gurɓataccen abu da ayyana asalin.

Gano tushen gurɓatattun abubuwa, kamar ragowar magungunan kashe qwari, karafa masu nauyi da sauran gurɓata kamar su polycyclic aromatic hydrocarbons (PAHs), shine matakin farko na sarrafa gurɓatawa. Yin feshin sinadarai na roba kai tsaye a cikin gonakin shayi, da kuma tukin iska da ake yi a kusa da lambunan shayi, su ne babban tushen ragowar maganin kashe qwari a cikin shayi[4]. Karafa masu nauyi na iya taruwa a cikin shayi kuma su haifar da guba, wanda galibi ana samun su ne daga kasa, taki da yanayi[5-7]. Dangane da sauran gurbatar yanayi da ke bayyana a cikin shayi ba zato ba tsammani, yana da wuya a gano shi saboda hadaddun hanyoyin samar da shayin da suka hada da shuka, sarrafawa, kunshin, ajiya da sufuri. PAHs a cikin shayi sun fito ne daga jibge abubuwan shaye-shaye na abin hawa da kona man da ake amfani da su yayin sarrafa ganyen shayi, kamar itacen wuta da gawayi[8-10].

A lokacin kona gawayi da itacen wuta, ana samun gurɓatattun abubuwa kamar carbon oxides[11]. A sakamakon haka, yana da sauƙi ga ragowar waɗannan gurɓatattun abubuwan da aka ambata a sama su faru a cikin samfuran da aka sarrafa, kamar hatsi, kayan da aka sha da kuma kifin cat, a cikin matsanancin zafin jiki, suna yin barazana ga lafiyar ɗan adam[12,13]. PAHs da aka haifar ta hanyar konewa sun samo asali ne daga rashin daidaituwa na PAHs da ke kunshe a cikin man fetur kanta, yanayin zafi mai zafi na mahadi masu kamshi da haɗin kai tsakanin free radicals [14]. Zazzaɓin ƙonewa, lokaci, da abun ciki na oxygen sune mahimman abubuwan da ke shafar canjin PAHs. Tare da karuwar zafin jiki, abubuwan da ke cikin PAH sun fara karuwa sannan kuma sun ragu, kuma ƙimar mafi girma ta faru a 800 ° C; Abubuwan da ke cikin PAHs sun ragu sosai don ganowa tare da ƙara lokacin konewa lokacin da yake ƙasa da iyaka da ake kira 'lokacin iyaka', tare da haɓaka abun ciki na oxygen a cikin iska mai ƙonewa, fitar da PAHs ya ragu sosai, amma rashin iskar oxygen da bai cika ba zai samar da OPAHs da sauran abubuwan da aka samo [15] -17].

9,10-Anthraquinone (AQ, CAS: 84-65-1, Fig. 1), wani nau'in oxygen-dauke da PAHs [18], ya ƙunshi nau'i-nau'i guda uku. An jera shi a matsayin mai yiwuwa carcinogen (Group 2B) ta Hukumar Bincike kan Ciwon daji ta Duniya a cikin 2014[19]. AQ na iya guba zuwa topoisomerase II cleavage complex da kuma hana hydrolysis na adenosine triphosphate (ATP) ta DNA topoisomerase II, haifar da DNA ninki biyu karya karya, wanda ke nufin cewa dogon lokaci daukan hotuna a karkashin AQ-dauke da yanayi da kuma kai tsaye lamba tare da babban matakin AQ. na iya haifar da lalacewar DNA, maye gurbi da ƙara haɗarin ciwon daji[20]. A matsayin mummunan tasiri akan lafiyar ɗan adam, Ƙungiyar Tarayyar Turai ta saita iyakar iyakar AQ (MRL) na 0.02 mg/kg a cikin shayi. Dangane da karatunmu na baya, an ba da shawarar ajiyar AQ a matsayin babban tushe yayin shukar shayi[21]. Hakanan, dangane da sakamakon gwaji a sarrafa koren shayi na Indonesiya, a bayyane yake cewa matakin AQ ya canza sosai kuma an ba da shawarar hayaki daga kayan aiki a matsayin ɗayan manyan dalilai[22]. Koyaya, ainihin asalin AQ a cikin sarrafa shayi ya kasance mai wuyar gaske, kodayake an ba da shawarar wasu hasashe na hanyar sinadarai na AQ [23,24], wanda ke nuna cewa yana da matuƙar mahimmanci don tantance mahimman abubuwan da ke shafar matakin AQ a sarrafa shayi.

Hoto 1. Tsarin sinadarai na AQ.

Idan aka ba da bincike kan samuwar AQ yayin konewar kwal da kuma yuwuwar barazanar mai a cikin sarrafa shayi, an gudanar da gwajin kwatancen don bayyana tasirin sarrafa tushen zafi akan AQ a cikin shayi da iska, ƙididdigar ƙima akan canje-canjen abun ciki na AQ. a matakai daban-daban na sarrafawa, waɗanda ke taimakawa don tabbatar da ingantaccen asalin, yanayin faruwa da matakin gurɓataccen AQ a sarrafa shayi.

SAKAMAKO
Tabbatar da hanyar
Idan aka kwatanta da bincikenmu na baya[21], an haɗa hanyar cire ruwa-ruwa kafin allura zuwa GC-MS/MS don inganta hankali da kiyaye maganganun kayan aiki. A cikin siffa 2b, hanyar da aka inganta ta nuna gagarumin ci gaba a cikin tsarkakewa na samfurin, mai narkewa ya zama mai sauƙi a launi. A cikin siffa 2a, cikakken bakan scan (50-350 m/z) ya kwatanta cewa bayan tsarkakewa, layin tushe na bakan MS ya ragu a fili kuma an sami ƙarancin kololuwar chromatographic, yana nuna cewa an cire babban adadin mahadi masu shiga tsakani bayan ruwa-ruwa hakar.

Hoto 2. (a) Cikakken bakan samfurin kafin da bayan tsarkakewa. (b) Tasirin tsarkakewa na ingantacciyar hanya.
Tabbatar da hanyar, ciki har da layi, dawo da, iyakar ƙididdigewa (LOQ) da tasirin matrix (ME), an nuna su a cikin Table 1. Yana da gamsarwa don samun layi tare da ƙididdiga na ƙaddara (r2) sama da 0.998, wanda ya kasance daga 0.005. zuwa 0.2 mg / kg a cikin matrix shayi da kuma acetonitrile ƙarfi, kuma a cikin samfurin iska tare da kewayon 0.5 zuwa 8 μg / m3.

An ƙididdige farfadowa na AQ a cikin ƙididdiga guda uku tsakanin ma'auni da ainihin ƙididdiga a cikin busassun shayi (0.005, 0.02, 0.05 mg / kg), sabon shayi na shayi (0.005, 0.01, 0.02 mg / kg) da samfurin iska (0.5, 1.5, 3). μg/m3). Farfadowar AQ a cikin shayi ya fito daga 77.78% zuwa 113.02% a busasshen shayi kuma daga 96.52% zuwa 125.69% a cikin ruwan shayi, tare da RSD% ƙasa da 15%. Farfadowa na AQ a cikin samfuran iska ya tashi daga 78.47% zuwa 117.06% tare da RSD% ƙasa da 20%. An gano mafi ƙasƙanci spiked taro a matsayin LOQ, wanda ya kasance 0.005 mg/kg, 0.005 mg/kg da 0.5 μg/m³ a shayi harbe, bushe shayi da kuma iska samfurori, bi da bi. Kamar yadda aka jera a cikin Table 1, matrix na busassun shayi da shayi na shayi sun ɗan ƙara yawan amsawar AQ, wanda ke haifar da ME na 109.0% da 110.9%. Dangane da matrix na samfuran iska, ME ya kasance 196.1%.

Matakan AQ yayin sarrafa koren shayi
Tare da manufar gano illolin hanyoyin zafi daban-daban kan shayi da yanayin sarrafa, an raba buhunan ganyayen ganye zuwa wasu takamaiman rukuni biyu kuma an sarrafa su daban a cikin tarurrukan sarrafa abubuwa guda biyu a cikin kamfani ɗaya. An ba wa wata ƙungiya wuta, ɗayan kuma da gawayi.

Kamar yadda aka nuna a cikin hoto 3, matakin AQ tare da wutar lantarki kamar yadda tushen zafi ya kasance daga 0.008 zuwa 0.013 mg / kg. A lokacin aikin gyarawa, bushewar ganyen shayin da aka yi ta hanyar sarrafawa a cikin tukunya mai zafin jiki ya haifar da karuwar kashi 9.5 cikin 100 na AQ. Sa'an nan kuma, matakin AQ ya kasance yayin aikin mirgina duk da asarar ruwan 'ya'yan itace, yana nuna cewa tsarin jiki bazai tasiri matakin AQ a cikin sarrafa shayi ba. Bayan matakan bushewa na farko, matakin AQ ya ƙaru kaɗan daga 0.010 zuwa 0.012 mg / kg, sannan ya ci gaba da tashi zuwa 0.013 mg / kg har zuwa ƙarshen sake bushewa. PFs, waɗanda suka nuna mahimmanci a cikin kowane mataki, sun kasance 1.10, 1.03, 1.24, 1.08 a cikin gyarawa, mirgina, bushewa na farko da sake bushewa, bi da bi. Sakamakon PFs ya nuna cewa sarrafawa a ƙarƙashin makamashin lantarki yana da ɗan tasiri akan matakan AQ a cikin shayi.

Hoto 3. Matsayin AQ yayin sarrafa koren shayi tare da wutar lantarki da gawayi azaman tushen zafi.
A cikin yanayin kwal a matsayin tushen zafi, abun ciki na AQ ya karu sosai yayin sarrafa shayi, yana tashi daga 0.008 zuwa 0.038 mg/kg. 338.9% AQ sun karu a cikin hanyar gyarawa, sun kai 0.037 mg / kg, wanda ya wuce MRL na 0.02 mg / kg da Tarayyar Turai ta kafa. Yayin matakin mirgina, matakin AQ har yanzu ya karu da 5.8% duk da kasancewa nesa da injin gyarawa. A farkon bushewa da sake bushewa, abun cikin AQ ya ƙaru kaɗan ko ya ragu kaɗan. PFs da ke amfani da gawayi azaman tushen zafi a cikin gyarawa, bushewa na farko da sake bushewa sune 4.39, 1.05, 0.93, da 1.05, bi da bi.

Don ƙarin ƙayyade dangantakar dake tsakanin konewar kwal da gurɓataccen AQ, an tattara abubuwan da aka dakatar da su (PMs) a cikin iska a cikin tarurrukan da ke ƙarƙashin duka tushen zafi don kimanta iska, kamar yadda aka nuna a cikin siffa 4. Matsayin AQ na PMs tare da kwal kamar yadda aka nuna. tushen zafi ya kasance 2.98 μg/m3, wanda ya ninka sau uku fiye da na wutar lantarki 0.91 μg/m3.

Hoto 4. Matakan AQ a cikin yanayi tare da wutar lantarki da gawayi azaman tushen zafi. * Yana nuna mahimman bambance-bambance a cikin matakan AQ a cikin samfuran (p <0.05).

Matakan AQ yayin sarrafa shayin Oolong shayi, wanda galibi ana samarwa a Fujian da Taiwan, wani nau'in shayi ne mai ɗanɗano. Don ƙara ƙayyade ainihin matakan haɓaka matakin AQ da tasirin mai daban-daban, nau'in nau'in ganye iri ɗaya an sanya shi cikin shayin oolong tare da garwashi da iskar gas-lantarki azaman tushen zafi, lokaci guda. Ana nuna matakan AQ a cikin sarrafa shayin oolong ta amfani da hanyoyin zafi daban-daban a cikin siffa 5. Don sarrafa shayin oolong tare da matasan iskar gas-lantarki, yanayin matakin AQ yana raguwa ƙasa da 0.005 mg/kg, wanda yayi kama da na kore shayi. da wutar lantarki.

Hoto 5. Matsayin AQ yayin sarrafa shayin oolong tare da gauran iskar gas-lantarki da gawayi azaman tushen zafi.

Tare da kwal azaman tushen zafi, matakan AQ a cikin matakai biyu na farko, bushewa da yin kore, sun kasance da gaske iri ɗaya da gauran iskar gas-lantarki. Koyaya, hanyoyin da suka biyo baya har sai an daidaita su sun nuna rata ta faɗaɗa a hankali, a lokacin matakin AQ ya ƙaru daga 0.004 zuwa 0.023 mg/kg. Matsayin da ke cikin matakan mirgina ya ragu zuwa 0.018 mg/kg, wanda zai iya zama saboda asarar ruwan shayi da ke ɗauke da wasu gurɓataccen AQ. Bayan matakin mirgina, matakin a cikin matakin bushewa ya karu zuwa 0.027 mg / kg. A cikin bushewa, yin kore, gyarawa, mirgina da bushewa, PFs sun kasance 2.81, 1.32, 5.66, 0.78, da 1.50, bi da bi.

Abin da ya faru na AQ a cikin samfuran shayi tare da tushen zafi daban-daban

Don ƙayyade abubuwan da ke tattare da abun ciki na AQ na shayi tare da maɓuɓɓugar zafi daban-daban, samfuran shayi na 40 daga wuraren bitar shayi ta amfani da wutar lantarki ko kwal kamar yadda aka yi nazarin hanyoyin zafi, kamar yadda aka nuna a cikin Tebu 2. Idan aka kwatanta da yin amfani da wutar lantarki azaman tushen zafi, kwal yana da mafi girma. ƙididdigar masu binciken (85.0%) tare da matsakaicin matakin AQ na 0.064 mg / kg, yana nuna cewa yana da sauƙi don haifar da gurɓataccen AQ ta hanyar tururi da aka samar ta hanyar konewa, kuma an lura da adadin 35.0% a cikin samfurori na kwal. Mafi mahimmanci, wutar lantarki tana da mafi ƙarancin bincike da ƙimar ƙimar 56.4% da 7.7% bi da bi, tare da matsakaicin abun ciki na 0.020 mg/kg.

TATTAUNAWA

Dangane da PFs yayin aiki tare da nau'ikan nau'ikan zafi guda biyu, a bayyane yake cewa gyare-gyare shine babban matakin da ya haifar da haɓaka matakan AQ a cikin samar da shayi tare da kwal da sarrafawa a ƙarƙashin makamashin lantarki yana da ɗan tasiri akan abun ciki na AQ. cikin shayi. A lokacin sarrafa koren shayi, konewar kwal yana haifar da hayaki mai yawa a cikin tsarin gyarawa idan aka kwatanta da tsarin dumama wutar lantarki, yana nuna cewa watakila tururi shine tushen tushen gurɓataccen AQ daga tuntuɓar harbe-harben shayi nan take a cikin sarrafa shayi, kama da tsarin fallasa a ciki. samfuran barbecue da aka kyafaffen[25]. Ƙara dan kadan a cikin abun ciki na AQ a yayin da ake yin mirgina ya nuna cewa hayaƙin da ya haifar da konewar kwal ba kawai ya shafi matakin AQ ba a lokacin matakin gyarawa, amma har ma a cikin yanayin sarrafawa saboda yanayin yanayi. An kuma yi amfani da gawayi azaman tushen zafi a farkon bushewa da sake bushewa, amma a cikin waɗannan matakai biyu abun cikin AQ ya ƙaru kaɗan ko ya ragu kaɗan. Ana iya yin bayanin hakan ta hanyar busar da iska mai zafi da aka rufe ta nisanta shayi daga hayakin da konewar kwal ke haifarwa[26]. Don tantance tushen gurɓataccen gurɓataccen abu, an yi nazarin matakan AQ a cikin yanayi, wanda ya haifar da babban rata tsakanin tarurrukan biyu. Babban dalilin wannan shi ne cewa kwal da aka yi amfani da shi a cikin gyaran gyare-gyare, bushewa na farko da sake bushewa zai haifar da AQ a lokacin konewa bai cika ba. Wadannan AQ sun kasance a cikin ƙananan barbashi na daskararru bayan konewar kwal kuma sun watse a cikin iska, suna haɓaka matakan gurɓataccen AQ a cikin yanayin bita[15]. A tsawon lokaci, saboda babban yanki na musamman da kuma ƙarfin tallan shayi, waɗannan ɓangarori sun zauna a saman ganyen shayin, wanda ya haifar da haɓakar AQ a cikin samarwa. Sabili da haka, ana tunanin konewar kwal shine babbar hanyar da ke haifar da gurɓataccen gurɓataccen AQ a cikin sarrafa shayi, tare da tururi shine tushen gurɓata.

Dangane da sarrafa shayin oolong, AQ ya ƙaru a ƙarƙashin sarrafawa tare da hanyoyin zafi guda biyu, amma bambanci tsakanin hanyoyin zafi biyu yana da mahimmanci. Sakamakon ya kuma nuna cewa kwal a matsayin tushen zafi ya taka muhimmiyar rawa wajen haɓaka matakin AQ, kuma an ɗauki gyare-gyare a matsayin babban mataki na haɓaka gurɓataccen AQ a cikin sarrafa shayi na oolong dangane da PFs. A lokacin sarrafa shayin oolong tare da matasan iskar gas-lantarki a matsayin tushen zafi, yanayin matakin AQ yana raguwa ƙasa da 0.005 mg / kg, wanda yayi kama da na kore shayi tare da wutar lantarki, yana ba da shawarar cewa makamashi mai tsabta, kamar wutar lantarki da na halitta. gas, na iya rage haɗarin samar da gurɓataccen AQ daga sarrafawa.

Dangane da gwaje-gwajen samfur, sakamakon ya nuna cewa yanayin gurɓataccen AQ ya fi muni yayin amfani da gawayi a matsayin tushen zafi maimakon wutar lantarki, wanda zai iya zama saboda hayaƙin konewar kwal da ke haɗuwa da ganyen shayi da kuma tsayawa a kusa da wurin aiki. Duk da haka, ko da yake a bayyane yake cewa wutar lantarki ita ce tushen zafi mafi tsabta yayin sarrafa shayi, har yanzu akwai gurɓataccen AQ a cikin kayan shayi ta amfani da wutar lantarki a matsayin tushen zafi. Halin yana da kama da aikin da aka buga a baya wanda aka ba da shawarar amsawar 2-alkenals tare da hydroquinones da benzoquinones a matsayin hanyar sinadarai mai yuwuwar[23], za a bincika dalilan wannan a cikin bincike na gaba.

KAMMALAWA

A cikin wannan aikin, yuwuwar tushen gurɓacewar AQ a cikin kore da shayi oolong an tabbatar da su ta hanyar gwaje-gwajen kwatancen bisa ingantattun hanyoyin nazarin GC-MS/MS. Bincikenmu ya goyi bayan kai tsaye cewa babban tushen gurɓataccen matakan AQ shine hayaƙin da konewa ya haifar, wanda ba kawai ya shafi matakan sarrafawa ba har ma ya shafi yanayin bita. Ba kamar a cikin matakan juyawa da bushewa ba, inda canje-canje a matakin AQ ba su da tabbas, matakan da ke da hannu kai tsaye na gawayi da itacen wuta, irin su gyarawa, shine babban tsari wanda kamuwa da AQ ya tashi saboda yawan haɗuwa tsakanin shayi. da hayaki a lokacin wadannan matakai. Don haka, an ba da shawarar tsaftataccen mai kamar iskar gas da wutar lantarki a matsayin tushen zafi a sarrafa shayi. Bugu da kari, sakamakon gwajin ya kuma nuna cewa, idan babu hayaki da konewa ke haifarwa, akwai sauran abubuwan da ke taimakawa wajen gano AQ yayin sarrafa shayi, yayin da aka kuma lura da kananan adadin AQ a wajen taron tare da tsaftataccen mai, wanda ya kamata a kara bincike. a cikin bincike na gaba.

KAYA DA MATAKAI

Reagents, sunadarai da kayan aiki

An saya daidaitattun Anthraquinone (99.0%) daga Kamfanin Dr. Ehrenstorfer GmbH (Augsburg, Jamus). D8-Anthraquinone na ciki misali (98.6%) an saya daga C/D/N Isotopes (Quebec, Kanada). Anhydrous sodium sulfate (Na2SO4) da magnesium sulfate (MgSO4) (Shanghai, China). Kamfanin Wenzhou Organic Chemical Company (Wenzhou, China) ne ya kawo Florisil. An siyi takarda fiber-gilasi (90 mm) daga kamfanin Ahlstrom-munksjö (Helsinki, Finland).

Samfurin shiri

An sarrafa samfuran shayi na kore tare da gyarawa, mirgina, bushewa na farko da sake bushewa (ta amfani da kayan aiki da aka rufe), yayin da samfuran shayi na oolong aka sarrafa tare da bushewa, yin kore (rocking da tsayawa sabobin ganye a madadin), gyarawa, cushe mirgina, da bushewa. An tattara samfurori daga kowane mataki sau uku a 100g bayan haɗuwa sosai. An adana duk samfuran a -20 ° C don ƙarin bincike.

An tattara samfurori na iska ta takarda fiber gilashi (90 mm) ta amfani da matsakaicin matsakaiciyar samfurin (PTS-100, Qingdao Laoshan Electronic Instrument Company, Qingdao, China) [27], yana gudana a 100 L / min na 4 h.

An ɗora samfurori masu ƙarfi tare da AQ a 0.005 mg / kg, 0.010 mg / kg, 0.020 mg / kg don sabon shayi mai shayi, a 0.005 mg / kg, 0.020 mg / kg, 0.050 mg / kg don busassun shayi kuma a 0.012 mg / kg (0.5 µg / m3 don samfurin iska), 0.036 mg / kg (1.5 µg / m3 don iska mai iska), 0.072 mg / kg (3.0 μg / m3 don samfurin iska) don takarda tace gilashi, bi da bi. Bayan girgiza sosai, an bar duk samfurori na tsawon sa'o'i 12, sannan kuma cirewa da matakan tsaftacewa.

An samo abun ciki na danshi ta hanyar ɗaukar 20 g na samfurin bayan haɗuwa kowane mataki, dumama a 105 ° C na 1 h, sannan aunawa da maimaita sau uku da ɗaukar matsakaicin darajar da rarraba shi da nauyi kafin dumama.

Samfurin cirewa da tsaftacewa

Samfurin shayi: Ana cirewa da tsarkakewa na AQ daga samfuran shayi bisa ga hanyar da aka buga daga Wang et al. tare da daidaitawa da yawa[21]. A taƙaice, 1.5 g na samfuran shayi an fara haxa shi da 30 μL D8-AQ (2 mg / kg) kuma an bar su don tsayawa na minti 30, sannan a haɗe su da ruwa mai ɗorewa na 1.5 mL kuma a bar su tsaya na minti 30. 15 ml 20% acetone a cikin n-hexane an ƙara shi zuwa samfuran shayi kuma ana sonicated don 15 min. Sa'an nan kuma an yi amfani da samfurori tare da 1.0 g MgSO4 na 30 s, kuma an yi amfani da su don 5 min, a 11,000 rpm. Bayan an motsa shi zuwa 100 ml na flasks masu siffar pear, 10 ml na babban tsarin kwayoyin halitta an kwashe shi zuwa kusan bushewa a karkashin injin a 37 ° C. 5 ml 2.5% acetone a cikin n-hexane ya sake narkar da abin da aka cire a cikin flasks masu siffar pear don tsarkakewa. Gilashin gilashi (10 cm × 0.8 cm) ya ƙunshi daga ƙasa zuwa saman gilashin ulu da 2g florisil, wanda ke tsakanin layuka biyu na 2 cm Na2SO4. Sannan 5 ml na 2.5% acetone a cikin n-hexane ya riga ya wanke ginshiƙi. Bayan loda maganin da aka sake narkar da shi, AQ ya sami haske sau uku tare da 5 ml, 10 ml, 10 ml na 2.5% acetone a cikin n-hexane. An mayar da eluates ɗin da aka haɗe zuwa kwalabe masu siffar pear kuma an kwashe su zuwa kusan bushewa a ƙarƙashin vacuum a 37 ° C. Sauran busassun an sake gyara shi tare da 1 ml na 2.5% acetone a cikin hexane sannan kuma tacewa ta hanyar tace girman pore 0.22 µm. Sa'an nan kuma an gauraye maganin da aka sake ginawa tare da acetonitrile a girman rabo na 1: 1. Bayan matakin girgiza, an yi amfani da subnatant don nazarin GC-MS/MS.

Samfurin iska: Rabin takardar fiber, wanda aka zubar da 18 μL d8-AQ (2 mg / kg), an nutsar da shi a cikin 15 mL na 20% acetone a cikin n-hexane, sannan sonicated don 15 min. An raba tsarin kwayoyin halitta ta hanyar centrifugation a 11,000 rpm na 5 min kuma an cire duk babban Layer a cikin flask mai siffar pear. An kwashe dukkan matakan kwayoyin halitta zuwa kusan bushewa a karkashin injin a zazzabi na 37 ° C. 5 ml na 2.5% acetone a cikin hexane ya sake narkar da kayan don tsarkakewa kamar yadda yake a cikin samfuran shayi.

GC-MS/MS bincike

Varian 450 gas chromatograph sanye take da Varian 300 tandem mai gano taro (Varian, Walnut Creek, CA, Amurka) an yi amfani da shi don yin nazarin AQ tare da software na MS WorkStation 6.9.3. Factor Varian Hudu shafi na capillary VF-5ms (30m × 0.25 mm × 0.25 μm) an yi amfani da shi don rabuwar chromatographic. Gas mai ɗaukar kaya, helium (> 99.999%), an saita shi a matsakaicin adadin 1.0 mL / min tare da iskar gas na Argon (> 99.999%). Zazzabi na tanda ya fara daga 80 ° C kuma an riƙe shi don 1 min; ya karu a 15 °C/min zuwa 240 °C, sannan ya kai 260 °C a 20 °C/min kuma a riƙe na 5min. Yanayin zafin jiki na tushen ion ya kasance 210 ° C, da kuma yanayin canja wuri na 280 ° C. Adadin allurar shine 1.0 μl. Ana nuna sharuɗɗan MRM a cikin Tebur 3.

Agilent 8890 gas chromatograph sanye take da Agilent 7000D sau uku quadrupole mass spectrometer (Agilent, Stevens Creek, CA, Amurka) an yi amfani dashi don nazarin tasirin tsarkakewa tare da software na MassHunter 10.1. Agilent J&W HP-5ms GC Column (30m × 0.25 mm × 0.25 μm) anyi amfani dashi don rabuwar chromatographic. An saita iskar gas mai ɗaukar nauyi, Helium (> 99.999%), a matsakaicin adadin 2.25 mL/min tare da iskar gas na Nitrogen (> 99.999%). An daidaita yanayin zafin EI ion a 280 ° C, daidai da zafin layin canja wuri. Yawan zafin jiki na tanda ya fara daga 80 ° C kuma an riƙe shi don 5 min; ya tashi da 15 ° C / min zuwa 240 ° C, sannan ya kai 280 ° C a 25 ° C / min kuma ana kiyaye shi don 5 min. Ana nuna sharuɗɗan MRM a cikin Tebur 3.

Binciken kididdiga
Abubuwan AQ a cikin sabobin ganye an gyara su zuwa busassun abun ciki ta hanyar rarraba abun cikin danshi don kwatantawa da nazarin matakan AQ yayin sarrafawa.

An kimanta canje-canjen AQ a samfuran shayi tare da software na Microsoft Excel da IBM SPSS Statistics 20.

An yi amfani da abubuwan sarrafawa don bayyana canje-canje a cikin AQ yayin sarrafa shayi. PF = Rl/Rf , inda Rf shine matakin AQ kafin matakin sarrafawa kuma Rl shine matakin AQ bayan matakin sarrafawa. PF yana nuna raguwa (PF <1) ko karuwa (PF> 1) a cikin ragowar AQ yayin takamaiman matakin sarrafawa.

ME yana nuna raguwa (ME <1) ko karuwa (ME> 1) don mayar da martani ga kayan aikin nazari, wanda ya dogara da rabon gangara na calibration a cikin matrix da sauran ƙarfi kamar haka:

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

Inda slopematrix shine gangara na lanƙwan daidaitawa a cikin matrix-matched sauran ƙarfi, slopesolvent shine gangara na lanƙwan daidaitawa a cikin sauran ƙarfi.

GABATARWA
Babban aikin Kimiyya da Fasaha na lardin Zhejiang (2015C12001) da Gidauniyar Kimiyya ta kasar Sin (42007354) ne suka tallafa wa wannan aikin.
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Hakkoki da izini
Haƙƙin mallaka: © 2022 ta marubuci(s). Babban Lasisin Matsakaicin Matsakaicin Ilimi, Fayetteville, GA. Wannan labarin buɗaɗɗen samun labarin labarin ne wanda aka rarraba ƙarƙashin lasisin Halayen Haɗin Kan Halittu (CC BY 4.0), ziyarci https://creativecommons.org/licenses/by/4.0/.
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