牛肉产地同位素与矿物元素指纹溯源技术研究
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摘要
稳定性同位素比率与矿物元素含量分析是用于食品产地溯源的有效方法之一。为了探索同位素与矿物元素指纹技术对我国牛肉产地溯源的可行性与有效性，本研究利用同位素比率质谱仪(IRMS)检测了不同地域来源的牛组织和饲料中δ~(13)C、δ~(15)N值，利用连续流动的同位素比率质谱仪(CF-IRMS)检测了牛尾毛中的δD值，利用等离子体质谱仪(ICP-MS)检测了脱脂牛肉中的~(206)Pb／~(207)Pb和~(208)Pb／~(207)Pb比值及Na、Mg、Al、K、Ca、V、Mn、Fe、Co、Ni、Cu、Zn、Ga、As、Se、Sr、Zr、Mo、Sn、Sb、Ba、Pb 22种矿物元素的含量，利用热电离质谱(TIMS)测定了脱脂牛肉中~(87)Sr／~(86)Sr比值，并借助SAS和DPS多元统计分析软件对检测结果进行了方差分析、多重比较分析、主成分分析、聚类分析和判别分析，系统研究了不同地域来源牛组织中同位素比率、矿物元素含量的差异，脱脂牛肉、粗脂肪、牛尾毛中碳、氮同位素组成的相关关系，各项同位素指标、矿物元素含量对牛肉产地的判别效果，以及它们与养殖环境的变化关系等。
此外，针对实际研究中牛肉样品采样困难及成本问题，用牛尾毛代替牛肉进行追溯研究的可能性，设计了以牛尾毛为采样材料的牛肉产地同位素溯源技术模型试验。比较了牛前部、中部、后部肌肉及心脏、肝脏、肺脏之间δ~(13)C、δ~(15)N值的差异，分析了牛尾毛生长的不同段位δ~(13)C、δ~(15)N值随喂养饲料的变化趋势，以及牛尾毛与肌肉组织、饲料中同位素的相关关系，探讨了利用牛尾毛为材料追溯牛肉产地及其生活史的可行性。
结果表明，不同地域来源的牛各组织中同位素比率与矿物元素含量有很大差异，并且有其各自的指纹特征：脱脂牛肉、粗脂肪、牛尾毛中碳、氮同位素呈极显著相关性，它们对牛肉产地的判别结果基本相同，均可用于牛肉产地溯源研究。在本研究的取样范围内，碳、氮、氘、铅和锶同位素对牛肉产地的判别效果由好到劣的次序依次为δ~(13)C＞δD＞~(206)Pb／~(207)Pb＞δ~(15)N＞~(208)Pb／~(207)Pb、~(87)Sr／~(86)Sr，判别较好的矿物元素指标依次为Se、Sr、Fe、Ni和Zn，两类指标组合能显著提高对牛肉产地的正确判别率。不同部位肌肉、不同内脏器官中碳、氮同位素无显著差异，即取样部位不影响碳、氮同位素比值的分析结果。饲料对牛组织中δ~(13)C、δ~(15)N值的影响远大于品种和个体的影响，尤其与主饲料成分密切相关，而且品种与个体对牛组织中δ~(15)N值的影响大于对δ~(13)C值的影响：在牛饲料中C_4植物与C_3植物比例差异较大的地域，δ~(13)C指标对牛肉产地的判别效果比较好：δ~(15)N指标可有效区分牧区与农区喂养的牛。牛组织中的δD值与地理条件密切相关，有随着纬度的增加而减小的趋势。牛组织对饲料中~(13)C的富集比例约在3‰左右，对~(15)N的富集比例约在3‰-5‰之间，饲料种类、牛品种、个体对富集比例均有一定影响；牛尾毛对~(13)C的富集比例大于脱脂牛肉，而对~(15)N的富集比例小于脱脂牛肉，脂肪组织对~(13)C有贫化作用。牛尾毛与脱脂肌肉、粗脂肪组织中碳、氮同位素呈极显著相关性，但随着饲料的改变，不同段位牛尾毛中碳、氮同位素与肌肉组织的相关性有很大差异；一般牛尾毛距离根部0—3cm与肌肉组织中δ~(13)C、δ~(15)N值相关性最高。
从上述研究结果中可以得出，同位素与矿物元素指纹技术对牛肉产地的追溯是行之有效的，以牛尾毛为研究材料追溯牛肉产地及其生活史是可行的。
The analysis of stable isotope ratio and mineral element content is one of effective methods for food geographical origin traceability. The objective of this article was to probe into the feasibility and validity using the analysis of isotope and minernal element fingerprint for cattle geographical origin traceability. Theδ~(13)C andδ~(15)N values of cattle tissues and feeds were determined using isotope ratio mass spectrometry (IRMS), the 8D values of cattle tail hair were determined using continuous flow-isotope ratio mass spectrometry (CF-IRMS), the ratio of ~(206)Pb/~(207)Pb, ~(208)Pb/~(207)Pb and the content of Na、Mg、Al、K、Ca、V、Mn、Fe、Co、Ni、Cu、Zn、Ga、As、Se、Sr、Zr、Mo、Sn、Sb、Ba、Pb were determined using the inductively coupled plasma mass spectrometry (1CP-MS), and the ratio of ~(87)Sr/~(86)Sr were determined using the thermal ionization mass spectrometry (TIMS). Then the data were performed using multivariate analysis including ANOVA, principal component analysis (PCA), cluster analysis (CA), linear discrimant analysis (LDA) with the help of SAS and DPS package for windows. The research were mainly focused on different analysis of stable isotope ratio and mineral element content of cattle from different regions, correlation analysis of carbon and nitrogen isotopic composion between de-fatted beef, crude fat and cattle tail hair, discrimant analysis using stable isotope ratios and mineral contents for cattle geographical origin assessment, and the relationship between stable isotope ratios, mineral contents of cattle tissues and their breeding environment.
In addition, the model experiment for cattle geographical origin traceability using stable isotope analysis was designed because of the difficulties in beef samples taking, higher cost, and the probability of using cattle tail hair instead of beef in the study of cattle traceability. The difference ofδ~(13)C,δ~(15)N values among muscle from foreside, central section, rump of cattle and cattle organs including heart, liver and lung were compared. The changing trends ofδ~(13)C,δ~(15)N values in different segment of cattle tail hair with the change of feeds composition were analyzed, and the correlationship between cattle tail hair, muscle, and feeds were studied to probe into the feasibility of tracing the cattle geographical origin and their feeding history using cattle tail hair instead of muscle.
The results showed that there were very significant differences according to region in the mean values of carbon, nitrogen, lead, hydrogen isotope ratios and minernal element content in the cattle tissues. The fingerprint of these indicators in cattle tissues from the different regions was unique. A significant correlationship was found in carbon and nitrogen isotope ratios between de-fatted beef, crude fat and cattle tail hair. The results of discrimant ananlysis were consistent using these indicators, which indicated that all of them could be used for cattle geographical origin treaceablity. The order of isotopic indicators wereδ~(13)C>δD>~(206)Pb/~(207)Pb>δ~(15)N>~(208)Pb/~(207)Pb and ~(87)Sr/~(86)Sr according to the rate of correct classification from high to low for cattle geographical origin, and the order of mineral element were Se>Sr>Fe>Ni>Zn according to the rate of correct clissification. Furthermore, the rate of correct classification could be improved by combination of these indicators. The model experiment showed that there was no significant difference inδ~(13)C andδ~(15)N values between muscles from different position and different organs, i.e. theδ~(13)C andδ~(15)N values of cattle tissues was not affected by sample from different position. The feeds, especially the staple feeds, had more effect on theδ~(13)C andδ~(15)N values of cattle tissues, while the variety and individual had less effect on them. The variety and individual had more effect onδ~(15)N thanδ~(13)C. Theδ~(13)C value in cattle tissues was very good in classifying the beef coming from the different regions where cattle feeds were significantly different in the proportion of C_4 and C3. Theδ~(15)N value was good to distinguish the cattle from the farm land and the pasture. The 8D in cattle tissues was closely related to the geographical conditions and was decreasing as the latitude was increasing. De-fatted beef and cattle tail hair enriched in ~(13)C to their feeds in the proportion of 3%o, and enriched in ~(15)N in the proportion of 3‰- 5‰, plus, cattle variety and individual had effect on the proportion of enrichment. Cattle tail hair enriched in ~(13)C much more than de-fatted beef, but less than de-fatted beef in ~(15)N, while fat tissue depleted in ~(13)C relative to feeds. Though a significant correlation was found between cattle tail hair, de-fatted beef and crude fat, there was a different correlation between cattle tail hair of different segment and muscle in carbon and nitrogen isotope ratio. The results showed that theδ~(13)C andδ~(15)N values of cattle tail hair in the segment which distance from root (0-3cm) was highly correlated with that of the muscle.
The above results showed that the technique of isotope and mineral element fingerprint was an effective method for cattle geographical origin traceability, and was feasible to trace the cattle geographical origin and their feeding histoty using cattle tail hair.
此外，针对实际研究中牛肉样品采样困难及成本问题，用牛尾毛代替牛肉进行追溯研究的可能性，设计了以牛尾毛为采样材料的牛肉产地同位素溯源技术模型试验。比较了牛前部、中部、后部肌肉及心脏、肝脏、肺脏之间δ~(13)C、δ~(15)N值的差异，分析了牛尾毛生长的不同段位δ~(13)C、δ~(15)N值随喂养饲料的变化趋势，以及牛尾毛与肌肉组织、饲料中同位素的相关关系，探讨了利用牛尾毛为材料追溯牛肉产地及其生活史的可行性。
结果表明，不同地域来源的牛各组织中同位素比率与矿物元素含量有很大差异，并且有其各自的指纹特征：脱脂牛肉、粗脂肪、牛尾毛中碳、氮同位素呈极显著相关性，它们对牛肉产地的判别结果基本相同，均可用于牛肉产地溯源研究。在本研究的取样范围内，碳、氮、氘、铅和锶同位素对牛肉产地的判别效果由好到劣的次序依次为δ~(13)C＞δD＞~(206)Pb／~(207)Pb＞δ~(15)N＞~(208)Pb／~(207)Pb、~(87)Sr／~(86)Sr，判别较好的矿物元素指标依次为Se、Sr、Fe、Ni和Zn，两类指标组合能显著提高对牛肉产地的正确判别率。不同部位肌肉、不同内脏器官中碳、氮同位素无显著差异，即取样部位不影响碳、氮同位素比值的分析结果。饲料对牛组织中δ~(13)C、δ~(15)N值的影响远大于品种和个体的影响，尤其与主饲料成分密切相关，而且品种与个体对牛组织中δ~(15)N值的影响大于对δ~(13)C值的影响：在牛饲料中C_4植物与C_3植物比例差异较大的地域，δ~(13)C指标对牛肉产地的判别效果比较好：δ~(15)N指标可有效区分牧区与农区喂养的牛。牛组织中的δD值与地理条件密切相关，有随着纬度的增加而减小的趋势。牛组织对饲料中~(13)C的富集比例约在3‰左右，对~(15)N的富集比例约在3‰-5‰之间，饲料种类、牛品种、个体对富集比例均有一定影响；牛尾毛对~(13)C的富集比例大于脱脂牛肉，而对~(15)N的富集比例小于脱脂牛肉，脂肪组织对~(13)C有贫化作用。牛尾毛与脱脂肌肉、粗脂肪组织中碳、氮同位素呈极显著相关性，但随着饲料的改变，不同段位牛尾毛中碳、氮同位素与肌肉组织的相关性有很大差异；一般牛尾毛距离根部0—3cm与肌肉组织中δ~(13)C、δ~(15)N值相关性最高。
从上述研究结果中可以得出，同位素与矿物元素指纹技术对牛肉产地的追溯是行之有效的，以牛尾毛为研究材料追溯牛肉产地及其生活史是可行的。
The analysis of stable isotope ratio and mineral element content is one of effective methods for food geographical origin traceability. The objective of this article was to probe into the feasibility and validity using the analysis of isotope and minernal element fingerprint for cattle geographical origin traceability. Theδ~(13)C andδ~(15)N values of cattle tissues and feeds were determined using isotope ratio mass spectrometry (IRMS), the 8D values of cattle tail hair were determined using continuous flow-isotope ratio mass spectrometry (CF-IRMS), the ratio of ~(206)Pb/~(207)Pb, ~(208)Pb/~(207)Pb and the content of Na、Mg、Al、K、Ca、V、Mn、Fe、Co、Ni、Cu、Zn、Ga、As、Se、Sr、Zr、Mo、Sn、Sb、Ba、Pb were determined using the inductively coupled plasma mass spectrometry (1CP-MS), and the ratio of ~(87)Sr/~(86)Sr were determined using the thermal ionization mass spectrometry (TIMS). Then the data were performed using multivariate analysis including ANOVA, principal component analysis (PCA), cluster analysis (CA), linear discrimant analysis (LDA) with the help of SAS and DPS package for windows. The research were mainly focused on different analysis of stable isotope ratio and mineral element content of cattle from different regions, correlation analysis of carbon and nitrogen isotopic composion between de-fatted beef, crude fat and cattle tail hair, discrimant analysis using stable isotope ratios and mineral contents for cattle geographical origin assessment, and the relationship between stable isotope ratios, mineral contents of cattle tissues and their breeding environment.
In addition, the model experiment for cattle geographical origin traceability using stable isotope analysis was designed because of the difficulties in beef samples taking, higher cost, and the probability of using cattle tail hair instead of beef in the study of cattle traceability. The difference ofδ~(13)C,δ~(15)N values among muscle from foreside, central section, rump of cattle and cattle organs including heart, liver and lung were compared. The changing trends ofδ~(13)C,δ~(15)N values in different segment of cattle tail hair with the change of feeds composition were analyzed, and the correlationship between cattle tail hair, muscle, and feeds were studied to probe into the feasibility of tracing the cattle geographical origin and their feeding history using cattle tail hair instead of muscle.
The results showed that there were very significant differences according to region in the mean values of carbon, nitrogen, lead, hydrogen isotope ratios and minernal element content in the cattle tissues. The fingerprint of these indicators in cattle tissues from the different regions was unique. A significant correlationship was found in carbon and nitrogen isotope ratios between de-fatted beef, crude fat and cattle tail hair. The results of discrimant ananlysis were consistent using these indicators, which indicated that all of them could be used for cattle geographical origin treaceablity. The order of isotopic indicators wereδ~(13)C>δD>~(206)Pb/~(207)Pb>δ~(15)N>~(208)Pb/~(207)Pb and ~(87)Sr/~(86)Sr according to the rate of correct classification from high to low for cattle geographical origin, and the order of mineral element were Se>Sr>Fe>Ni>Zn according to the rate of correct clissification. Furthermore, the rate of correct classification could be improved by combination of these indicators. The model experiment showed that there was no significant difference inδ~(13)C andδ~(15)N values between muscles from different position and different organs, i.e. theδ~(13)C andδ~(15)N values of cattle tissues was not affected by sample from different position. The feeds, especially the staple feeds, had more effect on theδ~(13)C andδ~(15)N values of cattle tissues, while the variety and individual had less effect on them. The variety and individual had more effect onδ~(15)N thanδ~(13)C. Theδ~(13)C value in cattle tissues was very good in classifying the beef coming from the different regions where cattle feeds were significantly different in the proportion of C_4 and C3. Theδ~(15)N value was good to distinguish the cattle from the farm land and the pasture. The 8D in cattle tissues was closely related to the geographical conditions and was decreasing as the latitude was increasing. De-fatted beef and cattle tail hair enriched in ~(13)C to their feeds in the proportion of 3%o, and enriched in ~(15)N in the proportion of 3‰- 5‰, plus, cattle variety and individual had effect on the proportion of enrichment. Cattle tail hair enriched in ~(13)C much more than de-fatted beef, but less than de-fatted beef in ~(15)N, while fat tissue depleted in ~(13)C relative to feeds. Though a significant correlation was found between cattle tail hair, de-fatted beef and crude fat, there was a different correlation between cattle tail hair of different segment and muscle in carbon and nitrogen isotope ratio. The results showed that theδ~(13)C andδ~(15)N values of cattle tail hair in the segment which distance from root (0-3cm) was highly correlated with that of the muscle.
The above results showed that the technique of isotope and mineral element fingerprint was an effective method for cattle geographical origin traceability, and was feasible to trace the cattle geographical origin and their feeding histoty using cattle tail hair.
引文
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