高效液相色谱法测定甜叶菊叶片中甜菊糖苷含量
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摘要
为检测化学调控下甜叶菊叶片中不同甜菊糖苷的含量,建立了快速分析甜叶菊叶片中3种甜菊糖苷含量的高效液相色谱测定方法。液相色谱分析采用Sepax HP-Amino(4.6×250 mm,5μm)色谱柱,柱温40℃,流速1.00 mL/min,流动相采用等度洗脱模式,流动相乙腈/水组成比为80∶20(v/v),检测器为紫外-可见检测器,检测波长210 nm,谱带宽度20 nm,狭缝宽度16 nm,进样体积4μL;样品采用超声波辅助热水提取,澄清石灰水去除杂质。结果表明液相色谱分析得出3种甜菊糖苷(甜菊糖苷、莱鲍迪苷C、莱鲍迪苷A)线性方程分别为Y=1.075X-1.853(r=0.999 9)、Y=1.302 X+1.136(r=0.999 5)和Y=1.117X+1.089(r=0.999 8),在0.05～5.00 mg/mL之间线性关系均表现良好,方法检测限分别为1.091μg/mL,3.062μg/mL和1.078μg/mL。所建方法简单灵敏,适合叶片样品快速检测。
In order to determine steviol glycosides in stevia rebaudiana leaves grown by chemicals control,a rapid determination of three steviol glycosides in stevia rebaudiana leaves was developed by using high performance liquid chromatography. The HPLC analysis was performed on a Sepax HP-Amino column(4.6×250 mm,5 μm)by isometric elution using acetonitrile/water(80 ∶ 20,v/v)as the mobile phase. The column temperature was 40 ℃,and the ?ow rate was 1.00 mL/min. The wavelength of UV-vis detector was 210 nm. The width of spectrum band and spectral slit were 20 nm and 16 nm respectively. The injection volume was 4 μL. The steviol glycosides in samples were extracted with hot water by ultrasonic assisted method and the extracts were pretreated by clear limewater to remove impurities. The results showed that the calibration curves developed for three steviol glycosides(stevioside,rebaudioside C and rebaudioside A)were Y=1.075 X-1.853(r=0.999 9),Y=1.302 X+1.136(r=0.999 5)and Y=1.117 X+1.089(r=0.999 8)respectively,and all had a good linear relationship within the concentration range of0.05 ～ 5.00 mg/mL. The limit of detection were 1.091 μg/mL,3.062 μg/mL and 1.078 μg/mL,respectively. This method was simple and sensitive for detecting steviol glycoisdes in leave products.
In order to determine steviol glycosides in stevia rebaudiana leaves grown by chemicals control,a rapid determination of three steviol glycosides in stevia rebaudiana leaves was developed by using high performance liquid chromatography. The HPLC analysis was performed on a Sepax HP-Amino column(4.6×250 mm,5 μm)by isometric elution using acetonitrile/water(80 ∶ 20,v/v)as the mobile phase. The column temperature was 40 ℃,and the ?ow rate was 1.00 mL/min. The wavelength of UV-vis detector was 210 nm. The width of spectrum band and spectral slit were 20 nm and 16 nm respectively. The injection volume was 4 μL. The steviol glycosides in samples were extracted with hot water by ultrasonic assisted method and the extracts were pretreated by clear limewater to remove impurities. The results showed that the calibration curves developed for three steviol glycosides(stevioside,rebaudioside C and rebaudioside A)were Y=1.075 X-1.853(r=0.999 9),Y=1.302 X+1.136(r=0.999 5)and Y=1.117 X+1.089(r=0.999 8)respectively,and all had a good linear relationship within the concentration range of0.05 ～ 5.00 mg/mL. The limit of detection were 1.091 μg/mL,3.062 μg/mL and 1.078 μg/mL,respectively. This method was simple and sensitive for detecting steviol glycoisdes in leave products.
引文
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[16]贺晓晗,王海增,郝华伟,等.普鲁兰多糖发酵液中色素及蛋白的脱除[J].食品工业科技,2015,36(7):138-143.
[17]孙蕊,贾鹏禹,靳革,等.高效液相色谱法测定木糖结晶母液中单糖[J].黑龙江八一农垦大学学报,2013,25(1):41-44.
[18]李瑞萍,袁琴,黄应平.硅胶色谱柱的亲水作用保留机理及其影响因素[J].色谱,2014,32(7):675-681.
[19]韩超,戴小军,雷根虎,等.强极性中药组分的亲水作用色谱保留行为[J].分析化学,2010,38(11):1615-1619.
[20]耿银银,许丽晓,欧定莲,等.亲水作用色谱法测定阿卡波糖含量[J].药物分析杂志,2017,37(5):818-825.品添加剂和配料展览会加剂生产应用技术展示会nts China 20193月18日-20日(上海),崧泽大道333号:www.cfaa.cn
[2]谢捷,刘小景,朱兴一,等.壳聚糖澄清甜叶菊水提液及其澄清机理探讨[J].食品科学,2011,32(20):1-6.
[3]王晓霞,王永胜,王涛,等.莱苞迪甙A的提取及精制工艺进展[J].化学通报,2016,79(9):811-816.
[4]孙大庆,李洪飞,张丽萍.柱色谱法提纯甜叶菊甙的工艺研究[J].农产品加工,2015(7):33-37.
[5]孙大庆,李洪飞,李良玉,等.模拟移动床色谱法分离甜叶菊甙的工艺研究[J].中国食品学报,2016,16(2):115-123.
[6]董海涛,孙宏义.NaCl胁迫对甜叶菊移栽苗生理生态特性的影响[J].冰川冻土,2015,37(2):538-544.
[7]陈叶,李番,马秀萍,等.5-氨基乙酰丙酸对甜叶菊产量和糖含量的影响[J].中国糖料,2017,39(2):36-38.
[8]朱吟吟,周凌.高效液相色谱法测定甜叶菊中的3个甜菊醇糖苷的含量[J].药物分析杂志,2014,34(1):184-189.
[9]张伟,王磊,钱建瑞,等.高效液相氨基柱和C18柱测定甜第二十三届中国国际食暨第二十九届全国食品添Food Ingredien展出时间:2019年展出地点:国家会展中心详情请登陆:菊糖含量方法对比[J].中国食品添加剂,2013(4):201-204.
[10]邹盛勤,姜琼.反相高效液相色谱法测定甜叶菊中A苷甜菊糖和甜菊苷含量[J].中国调味品,2014,39(6):121-123.
[11]张友财,俞坚,董雪瑞,等.HPLC法快速测定甜叶菊中瑞鲍迪苷A[J].食品工程,2014(4):60-63.
[12]汤其坤,王钰,吴跃进,等.近红外光谱法直接检测甜叶菊叶片甜菊糖苷模型建立[J].光谱学与光谱分析,2014,34(10):2719-2722.
[13]罗庆云,李小燕,邓瑞欣,等.酸性氧化铝用于甜叶菊甜菊糖苷分析样品前处理[J].中国糖料,2017,39(6):13-16,22.
[14]李山云,隋启君,杨琼芬,等.彩色马铃薯块茎色素初步分析[J].食品工业科技,2013,34(3):84-89.
[15]史一鸣,杨虹,李燕艳,等.亲水色谱法测定甜叶菊叶片中糖苷含量[J].食品研究与开发,2012,33(12):134-138.
[16]贺晓晗,王海增,郝华伟,等.普鲁兰多糖发酵液中色素及蛋白的脱除[J].食品工业科技,2015,36(7):138-143.
[17]孙蕊,贾鹏禹,靳革,等.高效液相色谱法测定木糖结晶母液中单糖[J].黑龙江八一农垦大学学报,2013,25(1):41-44.
[18]李瑞萍,袁琴,黄应平.硅胶色谱柱的亲水作用保留机理及其影响因素[J].色谱,2014,32(7):675-681.
[19]韩超,戴小军,雷根虎,等.强极性中药组分的亲水作用色谱保留行为[J].分析化学,2010,38(11):1615-1619.
[20]耿银银,许丽晓,欧定莲,等.亲水作用色谱法测定阿卡波糖含量[J].药物分析杂志,2017,37(5):818-825.品添加剂和配料展览会加剂生产应用技术展示会nts China 20193月18日-20日(上海),崧泽大道333号:www.cfaa.cn