摘要
建立测定畜肉中卡拉胶的高效液相色谱-质谱法。生鲜肉样品用1 mol/L盐酸处理,其中的卡拉胶降解生成[A-G4S]-等特征性寡糖片段,采用高效液相色谱-三重四极杆质谱进行定性和定量测定。可选择高分辨质谱用于确证。结果表明,对于正常畜肉样品不出现假阳性,对于标识添加卡拉胶的调理牛排等阳性样品均能有效识别。对于m/z 403>241离子对,检出限为20 mg/kg(卡拉胶/肉),线性范围为0.1～0.4 g/kg,线性相关系数在0.99以上,相对回收率为90%～120%。该方法前处理简便、检测指标明确、灵敏度高,可作为检测添加卡拉胶畜肉的有效手段。
This study aimed to establish a method for the determination of added carrageenan in livestock meat by high performance liquid chromatography-mass spectrometry(HPLC-MS). Carrageenan in fresh meat was digested with 1 mol/L hydrochloric acid, yielding characteristic oligosaccharide fragments such as [A-G4 S]-, which were detected by HPLC-triple quadrupole mass spectrometry. A high resolution mass spectrometer was employed for validation of the method. The results showed that no false positive results were found for natural meat samples, while positive samples such as steak labelled to contain carrageenan were identi?ed effectively by the proposed method. The limit of detection(LOD) of carrageenan was20 mg/kg(at m/z 403 > 241). A good linear relationship was observed in the concentration range of 0.1–0.4 g/kg with correlation coef?cients of more than 0.99. The relative recoveries were in the range of 90%–120%. In summary, this method, thanks to its simplicity, clear target and high sensitivity, could be used as an effective means to detect carrageenan-injected livestock meat.
引文
[1]PRAJAPATI V D,MAHERIYA P M,JANI G K,et al.Carrageenan:a natural seaweed polysaccharide and its applications[J].Carbohydrate Polymers,2014,105(1):97-112.DOI:10.1016/j.carbpol.2014.01.067.
[2]杨波.海洋硫酸半乳聚糖特异性降解、寡糖和糖脂的制备与序列分析及其寡糖芯片的构建[D].青岛:中国海洋大学,2002:2-3.
[3]郑炜,王君玮,赵思俊.生猪屠宰前违规使用卡拉胶的检测研究进展[J].中国动物检疫,2015,32(9):69-71.
[4]国家质量监督检验检疫总局.畜禽肉水分限量:GB 18394-2001[S].北京:中国标准出版社,2001.
[5]邓咏梅.关于我国猪肉水分限量的研究[J].肉类工业,2016(11):24-27.DOI:10.3969/j.issn.1008-5467.2016.11.008.
[6]王小军,佴梅.生猪屠宰注水监管中的问题分析[J].中国动物检疫,2014,31(9):31-32.
[7]HASSAN S S M,MEYERHOFF M E,BADR I H A,et al.Determination of carrageenan in food products using potentiometric polyion sensors[J].Electroanalysis,2002,14(6):439-444.
[8]王欣,王志永,陈利华,等.注胶肉糜的低场核磁弛豫特性及与近红外光谱技术判别效果的比较[J].食品与发酵工业2016,42(2):150-156.DOI:10.13995/j.cnki.11-1802/ts.201602027.
[9]吴艺影,章倩汝,韩剑众,等.基于低场核磁共振技术的注胶肉快速检测[J].肉类研究,2013,27(3):26-29.
[10]孟一,张玉华,许丽丹,等.近红外光谱技术对猪肉注水、注胶的快速检测[J].食品科学,2014,35(8):299-303.DOI:10.7506/spkx1002-6630-201408060.
[11]杨志敏.应用近红外光谱技术快速检测原料肉新鲜度及掺假的研究[D].杨凌:西北农林科技大学,2011:28-55.
[12]杨红菊,姜艳彬,侯东军,等.注胶肉的近红外光谱快速判别分析[J].肉类研究,2008,22(11):62-64.
[13]马悦欣,董双林,牟海津,等.κ-卡拉胶寡糖的酶解制备及其体外抗病毒活性[J].中山大学学报(自然科学版),2009,48(6):105-108.
[14]Z H O U G,S U N Y,X I N H,e t a l.I n v i v o a n t i t u m o r a n d immunomodulation activities of different molecular weight lambdacarrageenans from Chondrus ocellatus[J].Pharmacological Research,2004,50:47-53.
[15]EKEBERG D,KNUTSEN S H,SLETMOEN M.Negativeion electrospray ionization-mass spectrometry(ESI-MS)as a tool for analyzing structural heterogeneity in kappa-carrageenan oligosaccharides[J].Carbohydrate Research,2001,334:49-59.
[16]迟连利,于广利,任玮娜,等.κ-卡拉胶五糖的制备及结构研究[J].中国海洋药物,2002,21(3):22-24.DOI:10.3969/j.issn.1002-3461.2002.03.007.
[17]ANASTYUK S D,BARABANOVA A O,CORREC G,et al.Analysis of structural heterogeneity ofκ/β-carrageenan oligosaccharides from Tichocarpus crinitus by negative-ion ESI and tandem MALDI mass spectrometry[J].Carbohydrate Polymers,2011,86(2):546-554.DOI:10.1016/j.carbpol.2011.04.081.
[18]SUN Y J,YANG B Y,WU Y M,et al.Structural characterization and antioxidant activities ofκ-carrageenan oligosaccharides degraded by different methods[J].Food Chemistry,2015,178:311-318.DOI:10.1016/j.foodchem.2015.01.105.
[19]SUN Y J,LIU Y,JIANG K,et al.Electrospray ionization mass spectrometric analysis ofκ-carrageenan oligosaccharides obtained by degradation withκ-carrageenase from Pedobacter hainanensis[J].Journal of Agricultural and Food Chemistry,2014,62(11):2398-2405.DOI:10.1021/jf500429r.
[20]杨波.海洋硫酸半乳聚糖特异性降解、寡糖和糖脂的制备与序列分析及其寡糖芯片的构建[D].青岛:中国海洋大学,2009:84.
[21]BLAKEMORE W R,DAVIS S R,HRONCICH M M,et al.Carrageenan analysis.Part 1:characterisation of the carrageenan test material and stability in swine-adapted infant formula[J].Food Additives&Contaminants:Part A,2014,31:1661-1669.DOI:10.1080/19440049.2014.955536.
[22]BLAKEMORE W R,KWOK S K,HARDING N I.Carrageenan analysis:Part 2:quantification in swine-adapted infant formula[J].Food Additives and Contaminants:Part A,2014,31:1670-1672.DOI:10.1080/19440049.2014.955537.
[23]BLAKEMOREA W R,BRANT A F,BISSLAND J G,et al.Carrageenan analysis.Part 3:quantification in swine plasma[J].Food Additives&Contaminants:Part A,2014,31:1673-1677.DOI:10.1080/19440049.2014.955538.
[24]SHAHBAZI M,RAJABZADEH G,ETTELAIE R,et al.Kinetic study of kappa-carrageenan degradation and its impact on mechanical and structural properties of chitosan/kappa-carrageenan film[J].Carbohydrate Polymers,2016,142:167-176.DOI:10.1016/j.carbpol.2016.01.037.
[25]上官巧灵,陈海敏,严小军,等.离子对-反相高效液相色谱-三重四级杆质谱法测定λ-卡拉胶寡糖[J].中国海洋药物,2013,32(3):59-66.DOI:10.13400/j.cnki.cjmd.2013.03.008.
[26]高洋,陈海敏,徐继林,等.κ-卡拉胶寡糖的反相离子对-超高效液相色谱-四极杆-飞行时间质谱研究[J].分析化学研究报告,2009,37(11):1590-1595.
[27]CARAM-LELHAM N,SUNDEL?F L O,ANDERSSON T.Preparative separation of oligosaccharides fromκ-carrageenan,sodium hyaluronate,and dextran by SuperdexTM 30 prep.grade[J].Carbohydrate Research,1995,273:71-76.
[28]杨波,于广利,郝翠,等.系列ι-卡拉胶寡糖制备及其电喷雾串联质谱序列分析[J].高等学校化学学报,2010,31(2):303-308.
[29]PAULA G A,BENEVIDES N M B,CUNHA A P,et al.Development and characterization of edible films from mixtures of k-carrageenan,i-carrageenan,and alginate[J].Food Hydrocolloids,2015,47:140-145.DOI:10.1016/j.foodhyd.2015.01.004.
[30]陈欢欢,赵峡,栾晓红,等.电喷雾质谱在寡糖序列分析中的应用[J].高等学校化学学报,2015,36(1):1-8.DOI:10.7503/cjcu20140629.
