張艷東,程翠林,樊梓鸞,王振宇,*

(1.哈爾濱工業(yè)大學化工與化學學院,黑龍江哈爾濱 150090;2.東北林業(yè)大學林學院,黑龍江哈爾濱 150040)

植物精油的提取、成分分析以及抗氧化活性研究

張艷東1,程翠林1,樊梓鸞2,王振宇1,*

(1.哈爾濱工業(yè)大學化工與化學學院,黑龍江哈爾濱 150090;2.東北林業(yè)大學林學院,黑龍江哈爾濱 150040)

植物精油近年來得到越來越多的關(guān)注,人們對于精油的功效寄予厚望,對多種植物精油進行了研究,包括唇形科、菊科、桃金娘科以及姜科等植物的精油。不同植物精油在成分和含量上存在較大差異,因此在生物活性方面也存在一定差異。文章主要對近幾年來關(guān)于植物精油的提取、成分分析以及抗氧化活性的研究進行總結(jié)與概述,為精油的開發(fā)利用奠定研究基礎(chǔ)。

精油,提取,成分分析,抗氧化

植物精油因其在各領(lǐng)域的廣泛應(yīng)用而被大眾熟知,如化妝品、制藥以及食品工業(yè)等[1]。由于化學合成食品添加劑存在的潛在危害,人們把目光更多的投入到天然植物提取物上,植物精油就是越來越熱的研究之一。越來越多的方法被用于植物精油的提取以及成分分析,關(guān)于植物精油功能活性的研究也越來越多。本文將對近年來關(guān)于植物精油抗氧化活性的研究進行總結(jié)概述,希望對植物精油的開發(fā)利用起到一定的指導(dǎo)作用。

1 植物精油的提取

精油是指從植物的葉、花、根、莖或者果實中以一定工藝提取出來的具有揮發(fā)性的芳香類物質(zhì)。根據(jù)文獻記載,到2011年為止,世界上有17500種高等芳香植物可以用來提取精油,它們基本上都屬于少數(shù)幾個科,如桃金娘科、樟科、唇形科和菊科等[2]。提取植物精油的方法有很多,包括水蒸氣蒸餾法、蒸汽蒸餾法、索氏提取法、溶劑萃取法等傳統(tǒng)提取方法,以及超聲波輔助提取法、微波輔助提取法、超臨界流體萃取法、亞臨界流體萃取法等新方法。與新技術(shù)相比,傳統(tǒng)提取技術(shù)普遍存在能耗大、耗時長、得率低等缺陷[3-4]。除此之外,水蒸氣蒸餾和蒸汽蒸餾由于提取溫度高會導(dǎo)致精油中熱敏性成分的失活,水溶性成分溶解,而且對于親脂性成分的提取效果不佳[5-7]。而索氏提取與溶劑萃取存在有機溶劑殘留以及有機溶劑消耗過多等問題[8-9]。因此,為了解決傳統(tǒng)提取技術(shù)存在的缺陷,越來越多的新技術(shù)出現(xiàn)并且應(yīng)用在植物提取物的生產(chǎn)方面。

目前,超聲波輔助提取主要用于獲得親脂性和親水性的植物提取物[10]。與傳統(tǒng)提取技術(shù)相比,超聲波輔助提取技術(shù)是清潔的,而且超聲波的超聲空化效應(yīng)能夠加快溶劑滲透到植物細胞中,從而提高萃取效率并且防止提取物降解,相對來說減少了溶劑的使用量并且節(jié)約原料[11]。Aarti Singh等[4]采用水蒸氣蒸餾法和超聲波輔助提取法對桉樹葉子精油進行提取,實驗結(jié)果顯示兩種方法對該精油的得率分別為2.0%和2.6%,超聲波輔助提取法在得率上的優(yōu)勢不是特別突出,但是在提取時間上確實有較大優(yōu)勢:該實驗中水蒸氣蒸餾法的提取時間為5 h,而超聲波輔助提取法的提取時間只有30 min。

微波提取過程是通過高頻電磁波穿透介質(zhì)直達植物原料內(nèi)部實現(xiàn)的。微波輻射可以轉(zhuǎn)換成熱能迅速加熱,使細胞內(nèi)部溫度快速上升,細胞內(nèi)部水分大量蒸發(fā),從而對植物細胞壁產(chǎn)生壓力。當該壓力超過細胞壁承受能力時,細胞壁破裂,細胞內(nèi)的化學組分擴散到周圍的溶劑中,從而完成萃取的過程[12]。因此,與超聲波輔助提取相似,微波輔助提取也是清潔高效的新型提取技術(shù)。Burcu Uysal等[13]通過無溶劑微波輔助提取法和水蒸氣蒸餾法提取蕙花葉子精油,實驗結(jié)果顯示兩種方法對該精油的得率分別為2.5%和2.4%,雖然無溶劑微波輔助提取法在得率上沒有太大優(yōu)勢,但是在提取時間上無溶劑微波輔助提取法為20 min,而水蒸氣蒸餾法為3 h。

超臨界流體萃取是通過加熱、加壓的方式使溶劑(一般是CO2)達到超臨界狀態(tài),從而具有良好的溶劑特性,通過相似相容原理使植物細胞中的活性成分擴散到溶劑中,最后在降低溫度和壓強的情況下快速徹底的除去溶劑得到萃取物[14-15]。因此,超臨界流體萃取同樣是一項清潔高效的提取技術(shù),而且由于操作溫度較低,該技術(shù)對植物原料中的熱敏性成分具有良好的保護作用,因此超臨界流體萃取技術(shù)也是提取天然活性物質(zhì)的溫和方法。在植物精油萃取方面也得到了應(yīng)用[16-17]。Masoud Nejad-Sadeghi等[18]采用超臨界CO2萃取技術(shù)提取了青蘭地上部分的精油,在最佳實驗條件(萃取壓力240 bar,萃取溫度60 ℃,樣品粒度500 μm,CO2流速10 mL/s,萃取時間100 min)下該精油的最大得產(chǎn)率為2.72%。

由于植物組織和化學成分的差異,不同植物精油適合的提取方法也不盡相同,新型提取技術(shù)不一定在產(chǎn)率和生物活性等方面均優(yōu)于傳統(tǒng)提取技術(shù),因此在精油的提取過程中應(yīng)該通過完善的實驗設(shè)計探究最合適的提取方法。

2 植物精油成分分析

植物精油是多種組分的混合物,如單萜、倍半萜烯、醇類、酯類、醛類和酮類等,對于植物防御害蟲、食草動物、真菌以及細菌的侵襲有一定作用[22]。由于植物組織中各種化合物的理化性質(zhì)存在一定的差異,因此,選擇不同提取技術(shù)以及工藝參數(shù)對精油的成分具有較大的影響。為了得到品質(zhì)優(yōu)良的植物精油,成分分析是至關(guān)重要的。精油成分分析的方法有很多,例如薄層層析(TLC)、高效薄層色譜(HPTLC)、高效液相色譜(HPLC)、傅里葉變換紅外光譜(FTIR)、氣相色譜-氫火焰離子檢測器(GC-FID)、氣相色譜-質(zhì)譜聯(lián)用(GC-MS)和超臨界流體色譜(SFC)等等。

目前,在植物精油成分分析中GC-MS技術(shù)最為常用?；旌衔飿悠纷⑷肷V儀后,在色譜柱上分離,分離后的組分經(jīng)分子分離器除去載氣后,進入離子源,電離后,分子離子和碎片離子被加速并射向質(zhì)量分析器,從而達到根據(jù)樣品離子的質(zhì)量和強度進行定性定量和結(jié)構(gòu)分析的目的。Jie Chen等[23]通過GC-MS分析鑒定了水蒸氣蒸餾法得到的日本五針松松針精油,共鑒定出35種化合物,占精油總成分的96.34%。該松針精油富含單萜烯和倍半萜烯,主要為(+)-α-蒎烯(20.72%)、乙酸龍腦酯(19.74%)、大根香葉烯D(13.89%)和β-石竹烯(11.74%)。Ahmed M. Mustafa等[24]采用頂空固相微萃取(HS-SPME)耦合GC-MS技術(shù)提取分析野生種、栽培種和商業(yè)化黃龍膽根的精油,結(jié)果在22個樣品(野生種取自5個不同地區(qū),共11個樣品;四年栽培種取自同一地區(qū),共2個樣品;商業(yè)化黃龍膽根購于6個公司,共9個樣品)中共鑒定出150種化合物,主要為萜類化合物、醇類、醛類、酮類、酯類、烷類、脂肪酸衍生物和苯衍生物等等,不同樣品中各類物質(zhì)的含量略有不同。野生種和栽培種精油成分組成基本一致,但是與商業(yè)化品種差別較大。

HPTLC采用粒度分布特別窄的微粒硅劑(5～10 μm)制備成高效薄層板,用程序多級展開或園形展開技術(shù)使薄層色譜的靈敏度和分辨率大大提高;FTIR是由于物質(zhì)吸收電磁輻射后,分子振動-轉(zhuǎn)動能級的躍遷而產(chǎn)生的。FTIR光譜分析具有靈敏度高、試樣用量少、掃描時間極快、能分析各種狀態(tài)的試樣等特點,因此常用于物質(zhì)的定性鑒別和定量分析。Aarti Singh等[4]采用HPTLC和FTIR對提取的藍桉葉子精油進行定性評估,采用GC-MS技術(shù)對分離的精油進行半定量分析,通過HPTLC指紋識別發(fā)現(xiàn)在254 nm處混合物實現(xiàn)最大程度的分離,通過GC-MS分析發(fā)現(xiàn)該精油中共含有53種成分。

由此可見,植物精油成分定性鑒別和定量分析的技術(shù)多種多樣,在應(yīng)用時應(yīng)該根據(jù)精油的特征進行選擇,但是可以肯定的一點是GC-MS技術(shù)在植物精油成分分析中的應(yīng)用十分普遍。

3 植物精油抗氧化活性研究

氧化應(yīng)激對人體健康的影響已經(jīng)被越來越多的人關(guān)注,并且進行了大量的研究,各種天然抗氧化劑的研發(fā)也越來越多[35,36]。植物作為食物和藥物使用已經(jīng)有幾千年的歷史,中國有很多藥食同源的植物[37]。這些藥食同源的植物大多含有促進人體健康或者緩解疾病的植物素,很多具有抗氧化活性,如多酚類物質(zhì),存在于大多數(shù)芳香植物中,能夠清除體內(nèi)的自由基,預(yù)防和治療由于氧化應(yīng)激而引起的疾病[38-40]。植物精油作為芳香植物的“獨有標志”,其抗氧化活性被認為是許多應(yīng)用的基礎(chǔ),如食物保藏、制藥、非傳統(tǒng)醫(yī)學和自然療法等[41-42]。近年來,由于合成抗氧化劑在使用上的限制以及人們對天然無毒害抗氧化劑興趣的增加,植物精油已經(jīng)成為天然抗氧化劑的潛在來源之一,因此催生了很多關(guān)于植物精油抗氧化活性的研究[43]。

Soumaya Bourgou等[44]采用水蒸氣蒸餾法提取了四種不同起源的艾草地上部分的精油并進行體外抗氧化研究。其DPPH自由基清除實驗中四種精油的IC50值分別為1300、1300、110和400 μg/mL,而同等條件下BHT的IC50值為16 μg/mL;總還原力實驗中四種精油的EC50值(吸光值達到0.5時的濃度)分別為2、2.9、2.6和1.2 mg/mL,而同等條件下抗壞血酸的EC50值為0.032 mg/mL,說明四種精油均具有一定的抗氧化活性并且存在明顯差異。M. C. Scur等[45]采用水蒸氣蒸餾法提取番石榴新鮮葉子中的精油并通過DPPH自由基清除實驗來研究其抗氧化活性。實驗結(jié)果顯示該精油濃度為100 mg/mL時在該實驗條件下對DPPH自由基清除率僅為16.19%,其IC50值為171.14 mg/mL,而同等條件下BHT對DPPH自由基清除率為95.84%,其IC50值為11.51 mg/mL。實驗表明番石榴新鮮葉子精油對DPPH自由基的抑制活性較差,表現(xiàn)出較低的抗氧化活性。Qiao-Hui Zeng等[46]通過精油對DPPH自由基、ABTS自由基的清除實驗以及FRAP實驗研究了水蒸氣蒸餾法提取的野生大薊和小薊地上部分精油的抗氧化活性。確定在最適條件下(大薊:提取時間6 h,提取溫度160 ℃,料液比1∶15;小薊:提取時間6 h,提取溫度140 ℃,料液比1∶10)提取出的大薊和小薊精油具有較高的FRAP值和有效的ABTS自由基清除能力,并且隨著濃度的增大,精油對DPPH自由基和ABTS自由基的清除率逐漸增大,FRAP值也逐漸增大,在50～800 μg/mL的濃度范圍內(nèi)呈現(xiàn)良好的劑效關(guān)系。

以上研究都是圍繞不同來源的植物精油進行的,包括菊科的艾草精油和大薊、小薊精油,桃金娘科的番石榴葉精油,姜科的山奈精油,小檗科的南天竹果實精油以及蘭科的金線蓮精油。這些研究充分證明了植物精油來源的廣泛性,同時也說明抗氧化活性對于植物精油具有一定的普遍性。這對于植物精油能否成為天然抗氧化劑的潛在來源具有重要意義,同時對于同科不同種屬植物精油的開發(fā)也具有一定的指導(dǎo)作用。

4 結(jié)論與展望

本文從植物精油的提取技術(shù)、成分分析以及抗氧化活性研究三個方面入手,著重介紹了近幾年對于植物精油抗氧化活性的研究現(xiàn)狀,為植物精油的開發(fā)與應(yīng)用提供了一定的指導(dǎo)。植物精油因其獨特的功效一直受到廣大消費者的熱捧,近年來對于植物精油抗氧化活性的研究也越來越多,但是從目前發(fā)表的文獻來看,科研工作者對于植物精油抗氧化的研究僅限于簡單的體外抗氧化研究,對于植物精油抗氧化作用機制的研究尚未完善。而且由于芳香植物種類的多樣性,植物精油的種類也存在多樣性,因此,植物精油在提取、成分鑒定以及功效機制等方面具有廣闊的研究空間。

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Extraction,component analysis and antioxidant activity of plant essential oils

ZHANG Yan-dong1,CHENG Cui-lin1,FAN Zi-luan2,WANG Zhen-yu1,*

(1.School of Chemistry and Chemical Engineering,Harbin Institute of Technology,Harbin 150090,China;2.School of Forestry,Northeast Forestry University,Harbin 150040,China)

Plant essential oils have getting more and more attentions in recent years. People expect much of the effects of essential oils. A variety of plant essential oils were studied,including labiatae,compositae,myrtle and zingiberaceae. There are significant differences in chemical composition and content between different plant essential oils,which results in different biological activity. Different plant essential oils have biggish difference in chemical composition and content. Therefore,there also exist differences in biological activity. The researches about extraction,component analysis and antioxidant activity of plant essential oils in recent years were summarized in this paper,and this can help to lay a research foundation for the development and utilization of plant essential oils.

essential oil;extraction;composition analysis;antioxidant

2016-08-25

張艷東(1990-)，女，博士研究生，主要從事植物活性物質(zhì)的分離和功能研究，E-mail:15604805165@163.com。

*通訊作者:王振宇(1957-)，男，博士，教授，研究方向：天然產(chǎn)物化學、極端環(huán)境營養(yǎng)，E-mail:wzy219000@yeah.net。

十三五國家重點研發(fā)計劃(2016YFC0500305)。

TS201.2

A

:1002-0306(2017)03-0390-05

10.13386/j.issn1002-0306.2017.03.069