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苦蕎制品中黃曲霉毒素的污染及控制
本書是研究蕎麥制品的學(xué)術(shù)專著。本書從苦蕎的生理特性、產(chǎn)品研發(fā)、黃曲霉毒素的污染現(xiàn)狀及有關(guān)規(guī)定、相應(yīng)的檢測(cè)方法、生產(chǎn)控制與質(zhì)量認(rèn)證的八個(gè)方面對(duì)苦蕎中黃曲霉毒素的污染相關(guān)情況進(jìn)行了論述,可供檢驗(yàn)部門、政府實(shí)驗(yàn)室和食品企業(yè)品管部門、第三方實(shí)驗(yàn)室的技術(shù)人員參考,也可為與食品安全檢驗(yàn)、相關(guān)的苦蕎食品生產(chǎn)加工企業(yè)提供指導(dǎo)。
前 言蕎麥屬蓼科蕎麥屬雙子葉植物,是我國(guó)傳統(tǒng)的糧食作物,也是國(guó)際糧農(nóng)組織公認(rèn)的優(yōu)秀糧藥兼用糧種。我國(guó)苦蕎的種植面積和產(chǎn)量均位居世界第一,但對(duì)于苦蕎食品的研發(fā)只在最近的十幾年才開(kāi)始緩緩起步,其發(fā)展前景十分廣闊。食用苦蕎有降血壓、血糖、血脂,改善微循環(huán)等作用,在高脂肪、高熱量的食品充斥著現(xiàn)代人的餐桌與購(gòu)物籃的時(shí)候,有“三降”之稱的苦蕎食品異軍突起,成為市場(chǎng)上最為熱銷的產(chǎn)品之一。然而,苦蕎和人們所熟知的花生、玉米等農(nóng)作物一樣,容易遭受黃曲霉毒素的污染,因此,苦蕎食品的生產(chǎn)與銷售受到國(guó)家或地區(qū)設(shè)置黃曲霉毒素限量要求的技術(shù)壁壘,嚴(yán)重制約了苦蕎食品貿(mào)易的發(fā)展。 黃曲霉毒素主要是由黃曲霉真菌、寄生曲霉菌等菌株產(chǎn)生的,是一種毒性很強(qiáng)的肝毒素,其毒性為氰化鉀的10倍、砒霜的68倍,可引起肝臟的急性或慢性損害,也被證實(shí)有致癌、致畸、致突變的“三致”作用。此外,黃曲霉在花生、玉米、苦蕎等農(nóng)產(chǎn)品中廣泛存在,從作物田間種植到食品的生產(chǎn)加工與銷售,最后到消費(fèi)者消費(fèi)的整個(gè)過(guò)程都可能受到黃曲霉毒素的污染,因此,黃曲霉毒素被世界衛(wèi)生組織(WHO)的癌癥研究機(jī)構(gòu)劃定為Ⅰ類致癌物。 目前,對(duì)黃曲霉毒素關(guān)注和研究的意義主要表現(xiàn)在:① 社會(huì)公眾、媒體、政府對(duì)食品安全越來(lái)越關(guān)注;② 黃曲霉毒素對(duì)人類健康的影響及造成的危害很大;③ 國(guó)際貿(mào)易中作為技術(shù)壁壘的調(diào)節(jié)杠桿;④ 在國(guó)際反恐和戰(zhàn)爭(zhēng)中,防止其成為具有大規(guī)模殺傷力的生化武器,等等。 本書編者都是四川省教育廳重點(diǎn)實(shí)驗(yàn)室——苦蕎麥綜合利用實(shí)驗(yàn)室的成員,多年來(lái)一直從事苦蕎食品的研究,深感有必要通過(guò)全面系統(tǒng)地總結(jié)在該領(lǐng)域的工作經(jīng)驗(yàn),編寫一本較全面介紹苦蕎中黃曲霉毒素污染和控制方面的書。本書從苦蕎的生理特性、產(chǎn)品研發(fā)、黃曲霉毒素的污染現(xiàn)狀及有關(guān)規(guī)定、相應(yīng)的檢測(cè)方法、生產(chǎn)控制與質(zhì)量認(rèn)證等方面對(duì)苦蕎中黃曲霉毒素的污染相關(guān)情況進(jìn)行了介紹,可供檢驗(yàn)部門、政府實(shí)驗(yàn)室、食品企業(yè)品管部門、第三方實(shí)驗(yàn)室的技術(shù)人員參考,也可為與食品安全檢驗(yàn)、苦蕎食品生產(chǎn)加工相關(guān)的企業(yè)提供指導(dǎo)。 本書在編寫過(guò)程中,參閱了許多國(guó)內(nèi)外有關(guān)專家學(xué)者的論著,深入西昌航飛苦蕎科技發(fā)展有限公司、正中食品有限公司等苦蕎企業(yè)生產(chǎn)一線,與生產(chǎn)管理質(zhì)檢部門相結(jié)合,希望能從生產(chǎn)與檢測(cè)技術(shù)上為苦蕎產(chǎn)業(yè)的發(fā)展提供一定參考價(jià)值。 在編寫過(guò)程中,編者盡可能采用了最新的資料,但由于編者學(xué)識(shí)水平有限,書中難免存在不當(dāng)之處甚至錯(cuò)誤,懇請(qǐng)專家、讀者批評(píng)指正。
編 者 2015年4月
林巧,理學(xué)碩士,副教授,F(xiàn)任西昌學(xué)院輕化工程學(xué)院辦公室主任,主講食品生物化學(xué)、食品分析與檢驗(yàn)、食品發(fā)酵與釀造、果蔬儲(chǔ)存與加工等多門課程,涼山州學(xué)術(shù)帶頭人后備人選,西昌學(xué)院學(xué)術(shù)帶頭人后備人選。主持了四川省教育廳涼山苦蕎制品中黃曲霉毒素B1監(jiān)測(cè)體系的構(gòu)建、建昌板鴨微生物區(qū)系的研究等多個(gè)科研項(xiàng)目。
目 錄第一章 緒 論···················································································································· 1 第一節(jié) 苦蕎的特性············································································································· 1 第二節(jié) 苦蕎的研究進(jìn)展····································································································· 10 第三節(jié) 苦蕎制品的開(kāi)發(fā)狀況及前景展望·············································································· 15 第二章 產(chǎn)黃曲霉毒素真菌································································································· 30 第一節(jié) 黃曲霉毒素··········································································································· 30 第二節(jié) 產(chǎn)黃曲霉毒素真菌································································································· 31 第三節(jié) 黃曲霉毒素的生物合成機(jī)理···················································································· 38 第三章 國(guó)內(nèi)外黃曲霉毒素的限量標(biāo)準(zhǔn)················································································· 43 第一節(jié) 世界各國(guó)對(duì)食品中黃曲霉毒素總量的限量要求·························································· 43 第二節(jié) 我國(guó)食品中黃曲霉毒素的污染現(xiàn)狀與限量標(biāo)準(zhǔn)·························································· 50 第四章 苦蕎中黃曲霉毒素的污染及風(fēng)險(xiǎn)評(píng)估······································································· 56 第一節(jié) 食品中黃曲霉毒素的危害分析與風(fēng)險(xiǎn)評(píng)估································································· 56 第二節(jié) 黃曲霉毒素的毒理機(jī)制與危害················································································· 59 第三節(jié) 苦蕎中成分與黃曲霉毒素的關(guān)系·············································································· 66 第四節(jié) 苦蕎中黃曲霉毒素的暴露評(píng)估················································································· 72 第五節(jié) 苦蕎食品中黃曲霉毒素的風(fēng)險(xiǎn)評(píng)估··········································································· 75 第五章 苦蕎制品黃曲霉毒素的檢測(cè)···················································································· 81 第一節(jié) 采樣和制樣··········································································································· 81 第二節(jié) 高效液相色譜法····································································································· 83 第三節(jié) 酶聯(lián)免疫吸附法····································································································· 88 第四節(jié) 薄層色譜法··········································································································· 93 第五節(jié) 熒光光度法··········································································································· 97 第六節(jié) 實(shí)驗(yàn)室控制········································································································· 100 第六章 苦蕎制品中黃曲霉毒素的控制··············································································· 115 第一節(jié) 苦蕎種植期的控制································································································ 115 第二節(jié) 苦蕎收獲期的控制································································································ 116 第三節(jié) 苦蕎采購(gòu)期的控制································································································ 118 第四節(jié) 苦蕎制品加工期的控制························································································· 120 第五節(jié) 苦蕎制品儲(chǔ)藏期的控制························································································· 122 第七章 苦蕎制品質(zhì)量品質(zhì)認(rèn)證························································································· 126 第一節(jié) 苦蕎茶生產(chǎn)基本加工工藝與品質(zhì)形成······································································ 126 第二節(jié) HACCP基本原理································································································· 127 第三節(jié) HACCP與其他質(zhì)量控制體系的關(guān)系······································································· 128 第四節(jié) 苦蕎茶生產(chǎn)良好操作規(guī)范······················································································ 130 第五節(jié) HACCP應(yīng)用于苦蕎茶中黃曲霉毒素的控制····························································· 139 第八章 苦蕎制品中黃曲霉毒素的去除··············································································· 149 第一節(jié) 采用物理方法去除苦蕎中的AFB1·········································································· 149 第二節(jié) 采用化學(xué)方法去除苦蕎中的AFB1·········································································· 151 第三節(jié) 采用生物方法去除苦蕎中的AFB1·········································································· 152 第四節(jié) 黃曲霉毒素去除實(shí)例···························································································· 156 附 錄····························································································································· 161 附錄A 食品中黃曲霉毒素B1的測(cè)定················································································· 161 附錄B 食品中黃曲霉毒素M1和B1的測(cè)定······································································· 174 附錄C 免疫親和層析凈化高效液相色譜法和熒光光度法······················································ 180 附錄D 食品中黃曲霉毒素B1,B2,G1,G2的測(cè)定··························································· 188
一、苦蕎的藥用價(jià)值 苦蕎是營(yíng)養(yǎng)豐富的糧食作物,也是很好的藥用作物?嗍w作為一種傳統(tǒng)中藥,在我國(guó)很多醫(yī)書上都有記載,且朝鮮、韓國(guó)、日本、尼泊爾、印度、俄羅斯及歐洲許多國(guó)家的人民都用苦蕎來(lái)治病!侗静菥V目》中記載:“苦蕎麥味苦性品寒、有益氣、續(xù)精神、利耳目、降氣寬腸、磨積滯、消熱中風(fēng)、脾積泄瀉的作用。”《齊名要術(shù)》記有“頭風(fēng)畏冷者,飽食而三日即愈,神效。其秸作薦,可辟臭蟲(chóng)蜈蚣,燒煙熏之亦效。其殼與黑豆皮、菊花裝枕,明目”《植物名實(shí)圖考》稱蕎麥“性能消積,俗呼凈腸草”?嗍w具有降血壓、降血糖、降血脂,改善微循環(huán)等作用,因此又稱“三降”食品。 (一)降血糖 哈爾濱醫(yī)科大學(xué)公共衛(wèi)生學(xué)院一項(xiàng)調(diào)查表明,主食蕎麥地區(qū)的人群,其血糖水平和糖尿病患病率明顯低于不食用蕎麥地區(qū)的人群。其中,主食蕎麥的居民高血糖水平(即高于正常值的血糖水平)的檢出率為1.6%;而非主食蕎麥的人群,高血糖的檢出率為7.33%,后者高于前者近4倍。主食蕎麥和不食用蕎麥地區(qū)的糖尿病患病率也相差約1倍,前者為1.88%,后者為3.84%。這一結(jié)論提示,蕎麥作為有效降低糖尿病發(fā)生的雜糧作物,應(yīng)該更多地出現(xiàn)在老百姓的餐桌上。 (二)降血脂 動(dòng)物實(shí)驗(yàn)表明,用苦蕎麥蛋白復(fù)合物飼料喂小鼠后,小鼠血液中的紅細(xì)胞SOD(超氧物歧化酶,具有對(duì)抗自由基的作用)活性提高了15.1%,CAT(過(guò)氧化氫酶,催化過(guò)氧化氫分解成氧和水)活性提高了13.4%,全血GSH-PX(谷胱甘肽過(guò)氧化物酶,具有抗氧化作用)阻止自由基生成的活性提高了23.2%,肝臟中的SOD,CAT和GSH-Px的活性都提高了19.2%。 (三)降血壓 苦蕎麥有穩(wěn)定血壓的作用,對(duì)中輕度的血壓偏高治療作用明顯。這是因?yàn)榭嗍w麥中含有一般食品中不含有的黃酮類化合物──蘆丁。蘆丁可以清洗血液,軟化血管,改善微循環(huán),同時(shí)還有很好的抗酸化作用,使高血壓患者的毛細(xì)血管的脆性得到恢復(fù),達(dá)到降血壓的功效。 (四)對(duì)心血管系統(tǒng)的作用 1. 對(duì)心肌的作用 苦蕎麥中富含的生物類黃酮主要成分是蘆丁,又名VP,其主要功效是軟化血管、改善微循環(huán)、清熱解毒、活血化疲、拔毒生肌、益氣提神,有降血糖和尿糖的作用。荷蘭米切爾·赫托格牡博士對(duì)805名老年男性膳食中牛物類黃酮的含量進(jìn)行了測(cè)定,結(jié)果發(fā)現(xiàn),每天攝入生物類黃酮的量不小于30 mg的組,比每天攝入生物類黃酮量不大于19 mg的組,死于冠心病的危險(xiǎn)率降低了50%。 2. 對(duì)血管的作用 苦蕎生物類黃酮具有多方面的生理功能,其中的槲皮素和桑色素等具有改善機(jī)體血管平滑肌的收縮和舒張功能;蘆丁具有擴(kuò)張血管,維持毛細(xì)血管的抵抗力,降低其通透性及脆性,促進(jìn)細(xì)胞增生和防止血細(xì)胞的凝集,以及降血脂、擴(kuò)張冠狀動(dòng)脈、增強(qiáng)冠狀動(dòng)脈血流量等作用。同時(shí),蕎麥粉中所含豐富的維生素有降低人體血脂和膽固醇的作用,是治療高血壓和心血管病的重要輔助食(藥)物。蕎麥粉中含有的一些微量元素(如鎂、鐵、銅、鉀等)對(duì)于心血管具有保護(hù)作用。 (五)抗氧化作用 近年來(lái)大量醫(yī)學(xué)研究說(shuō)明,人體的過(guò)早衰老,是由于體內(nèi)過(guò)多的氧自由基將正常細(xì)胞氧化破壞而造成。自由基還可能損害身體內(nèi)的酶類和蛋白質(zhì)等,使器官、機(jī)體加速衰老。研究表明不同濃度的苦蕎多糖能顯著地消除超氧陰離子自由基和羥基自由基,隨著苦蕎麥多糖濃度的升高其抗氧化性作用逐漸增強(qiáng),苦蕎多糖對(duì)消除超氧陰離子自由基的作用效果明顯高于對(duì)羥基自由基的清除效果?嗍w中含有豐富的硒元素,而硒又是谷胱甘肽過(guò)氧化酶(GSH-Px)的必需成分,該酶能將有害的過(guò)氧化物還原成無(wú)害的羥基化合物,從而保護(hù)細(xì)胞膜的結(jié)構(gòu)與功能。硒化合物是人體衰老自由基的捕獲劑,可修復(fù)脫氧核糖核酸,控制細(xì)胞分裂繁殖,因此具有抗衰老作用。另外苦蕎籽粒殼還富含原花青素,該色素具有較強(qiáng)的抗氧化清除自由基的能力。類似的多酚類如原兒茶酸,綠原酸在苦蕎中含量豐富,并且都能清除自由基,因此具有延緩衰老防止氧化的作用?嗍w麥中的蘆丁、槲皮素等黃酮類物質(zhì)特別是槲皮素可清除超氧陰離子、羥基自由基等自由基,提高自由基清除酶SOD、GSH-Px活力,降低脂質(zhì)過(guò)氧化水平?嗍w中還含有豐富的維生素C和維生素E,皆為有效的抗氧化物質(zhì),可清除機(jī)體自由基以及人體代謝產(chǎn)生的肝臟毒物、脂質(zhì)過(guò)氧化物,能維持人體內(nèi)分泌平衡,促進(jìn)細(xì)胞再生,具有防止衰老的作用?梢哉f(shuō)苦蕎抗衰老的保健特性已經(jīng)融在它的骨子里了,是現(xiàn)代生活特別是都市生活中不可多得的清純糧食。 (六)抗癌、防癌作用 苦蕎生物類黃酮所富含的槲皮素等黃酮類化合物主要通過(guò)3個(gè)途徑起到抗癌和防癌作用,即抗自由基作用,直接抑制癌細(xì)胞生長(zhǎng),抗致癌因子。韃靼蕎生物類黃酮所含的5-羥基黃酮是防癌、抗癌、治癌的優(yōu)選藥物?嗍w麥中含有其他谷類作物缺乏的硒,有利于防癌。硒在人體內(nèi)與金屬相結(jié)合形成一種不穩(wěn)定的“金屬-硒-蛋白質(zhì)”復(fù)合物,有助于排解人體中的有毒物質(zhì)(如鉛等)。中國(guó)疾病預(yù)防控制中心陳君石研究員宣布,經(jīng)過(guò)多年研究發(fā)現(xiàn),在各種有免疫調(diào)節(jié)功能的營(yíng)養(yǎng)素(包括VC,VA,鋅和鎂等)中,硒是唯一可以直接抗病毒的營(yíng)養(yǎng)素。 美國(guó)科學(xué)家經(jīng)過(guò)12年試驗(yàn)后證實(shí),每天補(bǔ)充200 μg硒,可使大腸癌發(fā)病率降低48%,肺癌發(fā)病率降低46%,前列腺癌發(fā)病率降低63%。硒是聯(lián)合國(guó)衛(wèi)生組織確定的人體必需的微量元素,而且是該組織目前唯一認(rèn)定的防癌抗癌元素。國(guó)內(nèi)外民學(xué)研究證實(shí),人體缺硒會(huì)造成重要器官的機(jī)能失調(diào),人體有40多種疾病與飲食缺硒有直接關(guān)系。 美國(guó)癌癥研究所醫(yī)學(xué)專家指出,適量的硒幾乎能防止一切癌變。美國(guó)倡導(dǎo)飲食硒的攝入量200 μg/(d·人)。我國(guó)因有71%的地區(qū)處于低硒或缺硒水平,中國(guó)營(yíng)養(yǎng)學(xué)會(huì)建議飲食硒的攝入量為50~250 μg/(d·人)。安全攝入量是400 μg/(d·人),而我國(guó)平均對(duì)硒的攝入量尚不能達(dá)到40 μg/(d·人)的水平。這主要是由于食物里的含硒量不是很高,如小麥的含硒量是34 μg/kg,大米的含硒量是51 μg/kg,而苦蕎麥的含硒量是430 μg/kg。國(guó)外食品硒的來(lái)源多為將無(wú)機(jī)硒添加在飼料中轉(zhuǎn)化,在食物鏈上為二級(jí)吸收,我國(guó)苦蕎麥硒為天然有機(jī)硒,以綠色植物為載體,所以十分珍貴。 (七)清除自由基的作用 自由基(free radical)是引起癌癥、衰老、心腦血管退變等疾病的罪惡之源。因?yàn)榭嗍w類黃酮物質(zhì)易被氧化,才使其具有清除自由基的功能。苦蕎生物類黃酮組成的分子結(jié)構(gòu)符合有效酚羥基理論,具有極強(qiáng)的自由基清除能力,它們具有5個(gè)羥基,可以充足地作為供氫體,使自由基還原,從而起到清除自由基的作用,達(dá)到防衰、抗癌、抗心腦血管病的目的。 (八)對(duì)骨組織的作用 苦蕎生物類黃酮可用于治療骨病和骨質(zhì)疏松等癥,其作用機(jī)理在于:(1)它既可抑制前列腺素E2(PGE2)的膠原蛋白合成增加,又能抑制PGE2的膠原蛋白合成減少,即抑制BH-脯氨酸進(jìn)入可消化的膠原蛋白和非膠原蛋內(nèi),并在低濃度PGE2時(shí)主要作用于非膠原蛋白的合成,高濃度PGE2主要作用于膠原蛋白合成,因此可以用于治療骨病;(2)它能提高甲狀腺對(duì)雌激素的敏感性,使甲狀腺C細(xì)胞分泌降鈣素的作用加強(qiáng),最終抑制骨再吸收而治療骨質(zhì)疏松;(3)它能抑制飲食中因缺鈣和VD而引起的骨密度和骨鈣含量的降低。 (九)護(hù)肝作用 苦蕎生物類黃酮對(duì)急慢性肝炎、肝硬化、脂肪肝以及因半乳糖胺等引起的中毒性肝損傷均有一定的療效。研究表明,苦蕎生物類黃酮對(duì)CCU所致肝臟的丙二醛(MDA肝脂質(zhì)過(guò)氧化終產(chǎn)物)含量的增高有明顯的抑制作用;可減輕肝損傷對(duì)谷胱甘肽(GSH)的消耗;能夠保護(hù)細(xì)胞結(jié)構(gòu)的完整性,防止肝細(xì)胞中的轉(zhuǎn)氨酶等水溶性酶因從細(xì)胞中漏出而活性升高?嗍w生物類黃酮的護(hù)肝機(jī)制最終在于它的抗氧化和抗自由基的作用。 (十)抗紫外線作用 苦蕎生物類黃酮所富含的槲皮素等黃酮類化合物是一種光屏蔽物,具有吸收紫外線的功能?杀Wo(hù)抗壞血酸等生物活性物質(zhì)免除氧化,保持了它們的活性,是一種極好的抗氧化劑,其螯合作用和基因表達(dá)出減少脂肪氧化變性的功能作用明顯,尤其是該黃酮對(duì)皮膚有保健作用以及抗炎癥作用,故有人稱之為“皮膚營(yíng)養(yǎng)素”。 (十一)其他保健作用 苦蕎還具有其他多種保健功能,如抗菌、抗病毒作用。研究證實(shí),苦蕎生物類黃酮所富含的槲皮素、桑色素、莰非醇有抑菌和抗病毒作用。臨床應(yīng)用黃酮,對(duì)各種原因引起的潰瘍有祛腐生肌、消炎鎮(zhèn)痛的奇特療效。對(duì)免疫系統(tǒng)的作用,苦蕎生物類黃酮能明顯有效地增加機(jī)體的非特異免疫功能和體液免疫功能,且能抗疲勞改善記憶?嗍w中多酚物質(zhì)對(duì)改善記憶力、預(yù)防老年癡呆癥有很好的效果,苦蕎醇提取物能夠延長(zhǎng)戊巴比妥鈉閾上的劑量條件下小鼠睡眠的時(shí)間,從而具有鎮(zhèn)靜催眠的功能,起到延長(zhǎng)催眠安神的保健功效。
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