分级净化结合气相色谱质谱联用法测定豆芽中10种植物生长调节剂
吴平谷等
摘 要 建立了豆芽10种植物生长调节剂的分级净化体系，
1 引 言
我国豆芽以作坊式生产为主，在豆芽生产过程中肆意添加植物生长调节剂催发豆芽生长常有发生，添加的植物生长调节剂主要有等。本研究针对豆芽生产过程中可能使用的10种植物生长调节剂，通过加标实验，根据其化学性质进行分级净化，采用气相色谱质谱法对该净化体系的进行了评价，获得较好的净化效果，有较大实际应用价值。
2 实验部分
2.1 仪器与试剂
3.5 样品分析
从杭州超市、农贸市场中采集豆芽各10份进行10种植物生长调节剂残留分析，主要检出CPA和吲哚乙酸，检出率
分别为50%和20%，含量范围为0.014～0.26 mg/kg，，CPA已经从GB27622011《食品安全国家标准 食品添加剂使用标准》中删除了在豆芽生产中使用的规定。吲哚乙酸是植物内源性生长素之一，可以促进豆芽杆的生长，未见豆芽中吲哚乙酸本底含量的报道，虽然吲哚乙酸对动物的毒性较低， 目前我国尚未规定吲哚乙酸在豆芽生产中应用。
References
1 SU FangFei， HU MengQing， MEI TianZi， YANG Bei， YANG ShuLong ， ZOU WeiYing， ZOU Ting， ZHANG DaLei. Journal of Nanchang University（Medical Science）， 2013， 53（1）： 13-15
苏芳菲 ， 胡梦青 ， 梅天资， 杨 蓓 ， 杨树龙 ， 邹惟莹 ， 邹 挺 ， 张大雷. 南昌大学学报（医学版）， 2013， 53（1）： 13-15
2 XU AiDong. China Vegetables， 2009 （8）： 1-6
徐爱东. 中国蔬菜， 2009 （8）： 1-6
3 WANG YiQian， ZHANG GuangHua， HE HongJu. Modern Instruments， 2012， 18（2 ）： 6-10
王一茜， 张广华， 何洪巨， 现代仪器， 2012， 18（2 ）： 6-10
4 GB/T232052008 Determination of 448 pesticides and related chemicals residues in tea LCMSMS method. National Standards of the People′s Republic of China
茶叶中448种农药多残留及相关化学品残留量的测定 液相色谱质谱法， 中华人民共和国国家标准 GB/T232052008
5 LIU JingJing， GONG Ping， ZHANG XiaoMei， WANG JianHua， WANG JingTang. Chinese Journal of Chromatography， 2012， 30（10）： 1012-1016
刘靖靖， 宫 萍， 张晓梅， 王建华， 王境堂. 色谱， 2012， 30（10）： 1012-1016
6 MOU YanLi，GUO DeHua，DING ZhuoPing，YI XiongHai. Chinese J. Anal. Chem.， 2013， 41（11）： 1640-1646
牟艳莉， 郭德华， 丁卓平， 伊雄海. 分析化学， 2013， 41（11）： 1640-1646
7 WU FengQi， JIN BaoHui， CHEN Bo， CHEN PeiJin， XIAO Feng. Chinese Agricultural Science Bulletin， 2010， 26（15）： 115119
吴凤琪， 靳保辉， 陈 波， 陈沛金， 肖 锋. 中国农学通报， 2010， 26（15）： 115-119
8 MOU YanLi， GUO DeHua， DING ZhuoPing， Journal of Instrumental Analysis， 2013， 32（8）： 935-940
牟艳利， 郭德华， 丁卓平. 分析测试学报， 2013， 32（8）： 935-940
9 Ma L， Zhang H， Xu W， He X， Yang L， Luo Y， Huang K. Food Anal. Methods， 2013， 6（3）： 941-951
10 Zhang F， Zhao P， Shan W， Gong Y， Jian Q， Pan C. Bull Environ Contam Toxicol， 2012， 89： 674-679
11 Anastasslades M， Lehotay S J， Stajnbaher D. J. AOAC International， 2003， 86（2）： 412-431
12 ZHAO YanSheng， DONG Ying， ZHANG Feng， YANG MinLi， FENG Feng， CHU XiaoGang Chinese J. Anal. Chem.， 2012， 40（2）： 249-256
赵延胜， 董 英， 张 峰， 杨敏莉， 冯 峰， 储晓刚. 分析化学， 2012， 40（2）： 249-256
13 WU PingGu， WANG Qiang， CHEN HuiHua， YING YongFei， ZHAO YongXin . Chinese J. Anal. Chem.， 2008， 36（11）： 1476-1482
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
摘 要 建立了豆芽10种植物生长调节剂的分级净化体系，
1 引 言
我国豆芽以作坊式生产为主，在豆芽生产过程中肆意添加植物生长调节剂催发豆芽生长常有发生，添加的植物生长调节剂主要有等。本研究针对豆芽生产过程中可能使用的10种植物生长调节剂，通过加标实验，根据其化学性质进行分级净化，采用气相色谱质谱法对该净化体系的进行了评价，获得较好的净化效果，有较大实际应用价值。
2 实验部分
2.1 仪器与试剂
3.5 样品分析
从杭州超市、农贸市场中采集豆芽各10份进行10种植物生长调节剂残留分析，主要检出CPA和吲哚乙酸，检出率
分别为50%和20%，含量范围为0.014～0.26 mg/kg，，CPA已经从GB27622011《食品安全国家标准 食品添加剂使用标准》中删除了在豆芽生产中使用的规定。吲哚乙酸是植物内源性生长素之一，可以促进豆芽杆的生长，未见豆芽中吲哚乙酸本底含量的报道，虽然吲哚乙酸对动物的毒性较低， 目前我国尚未规定吲哚乙酸在豆芽生产中应用。
References
1 SU FangFei， HU MengQing， MEI TianZi， YANG Bei， YANG ShuLong ， ZOU WeiYing， ZOU Ting， ZHANG DaLei. Journal of Nanchang University（Medical Science）， 2013， 53（1）： 13-15
苏芳菲 ， 胡梦青 ， 梅天资， 杨 蓓 ， 杨树龙 ， 邹惟莹 ， 邹 挺 ， 张大雷. 南昌大学学报（医学版）， 2013， 53（1）： 13-15
2 XU AiDong. China Vegetables， 2009 （8）： 1-6
徐爱东. 中国蔬菜， 2009 （8）： 1-6
3 WANG YiQian， ZHANG GuangHua， HE HongJu. Modern Instruments， 2012， 18（2 ）： 6-10
王一茜， 张广华， 何洪巨， 现代仪器， 2012， 18（2 ）： 6-10
4 GB/T232052008 Determination of 448 pesticides and related chemicals residues in tea LCMSMS method. National Standards of the People′s Republic of China
茶叶中448种农药多残留及相关化学品残留量的测定 液相色谱质谱法， 中华人民共和国国家标准 GB/T232052008
5 LIU JingJing， GONG Ping， ZHANG XiaoMei， WANG JianHua， WANG JingTang. Chinese Journal of Chromatography， 2012， 30（10）： 1012-1016
刘靖靖， 宫 萍， 张晓梅， 王建华， 王境堂. 色谱， 2012， 30（10）： 1012-1016
6 MOU YanLi，GUO DeHua，DING ZhuoPing，YI XiongHai. Chinese J. Anal. Chem.， 2013， 41（11）： 1640-1646
牟艳莉， 郭德华， 丁卓平， 伊雄海. 分析化学， 2013， 41（11）： 1640-1646
7 WU FengQi， JIN BaoHui， CHEN Bo， CHEN PeiJin， XIAO Feng. Chinese Agricultural Science Bulletin， 2010， 26（15）： 115119
吴凤琪， 靳保辉， 陈 波， 陈沛金， 肖 锋. 中国农学通报， 2010， 26（15）： 115-119
8 MOU YanLi， GUO DeHua， DING ZhuoPing， Journal of Instrumental Analysis， 2013， 32（8）： 935-940
牟艳利， 郭德华， 丁卓平. 分析测试学报， 2013， 32（8）： 935-940
9 Ma L， Zhang H， Xu W， He X， Yang L， Luo Y， Huang K. Food Anal. Methods， 2013， 6（3）： 941-951
10 Zhang F， Zhao P， Shan W， Gong Y， Jian Q， Pan C. Bull Environ Contam Toxicol， 2012， 89： 674-679
11 Anastasslades M， Lehotay S J， Stajnbaher D. J. AOAC International， 2003， 86（2）： 412-431
12 ZHAO YanSheng， DONG Ying， ZHANG Feng， YANG MinLi， FENG Feng， CHU XiaoGang Chinese J. Anal. Chem.， 2012， 40（2）： 249-256
赵延胜， 董 英， 张 峰， 杨敏莉， 冯 峰， 储晓刚. 分析化学， 2012， 40（2）： 249-256
13 WU PingGu， WANG Qiang， CHEN HuiHua， YING YongFei， ZHAO YongXin . Chinese J. Anal. Chem.， 2008， 36（11）： 1476-1482
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
吴平谷， 王 强， 沈向红， 陈慧华， 宋国良， 应永飞， 徐小民， 赵永信.分析化学， 2008， 36（11）： 1476-1482
Determination of 10 Plant Growth Regulators in Bean Sprouts by
Sequential CleaningGas ChromatographyMass Spectrometry
WU PingGu， TAN Yin， ZHANG Jin， WANG LiYuan， TANG Jun，
JIANG Wei， PAN XiaoDong， MA BingJie， NI ZhuNan， WANG TianJiao
（Zhejiang Provincial Center for Disease Control and Prevention， Hangzhou 310051， China）
Abstract A sequential cleanup method was developed for the quantification of 10 plant growth regulators in bean sprout by the gas chromatography/mass spectrometry （GC/MS）. The analytes were firstly extracted by the acided acetonitrile. Extraction was concentrated and redissovled by methanol. Then， it was divided to two aliquots. One of that was analyzed for 2，4Dbutyl ester and 2，4Dethyl ester after the purification by QuECHERS cartridge. Another one was treated by MCS solid phase extraction column including diverse eluting steps. After eluting by 5 mL methanol， composition 1 was obtain， concentrated， and methyl esterified by 10% boron trifluoride methanol solution. The treated extract was used for the determination of 4chlorophenoxy acetic acid， βnaphthyl acetic acid， 2，4dichlorophenoxy acetic acid， indole acetic acid and indole butyric acid. Composition 2 collected by eluting with 5 mL 5% amonium methanol was used for the determination of paclobutrazol， Kinetin， 6Benzylaminopurine. The cleanup procedures are designed according to different chemistry properties of these plant growth regulators. The results showed that after spiking of 0.01-0.1 mg/kg selected plant growth regulators， average recovery ranged from 70.0% to 93.2% and relative standard deviation were 5.2%-12.3%. Limit of quantification （LOQ S/N≥10） and limit of detection （LOD S/N≥3） were 0.01-0.025 mg/kg and 0.003-0.008 mg/kg respectively. The developed purification method is easy， fast and accurate， and can be applied to routine test of plant growth regulators in bean sprout.
Keywords Bean sprout； Plant growth regulator； Sequential cleaning； Gas chromatographymass spectrometry
（Received 3 February 2014； accepted 17 March 2014）
