混合固相萃取—HPLC测定豆芽中5种植物生长调节剂

Determination of 5 kinds of plant growth regulator in bean sprout by using mixed solid phase extraction column coupled with high performance liquid chromatography

目的建立基于自制混合型固相萃取柱净化样品-高效液相色谱同时测定豆芽中6-苄基腺嘌呤、异戊烯基腺嘌呤、赤霉素、4-氟苯氧乙酸、4-氯苯氧乙酸的方法。方法样品经20 ml乙腈提取,盐析离心,通过自制固相萃取柱净化,经Spursil C18-EP色谱柱分离,乙腈和0.5%磷酸为流动相,紫外检测器检测,检测波长为209 nm,外标法定量。结果 5种植物生长调节剂在0.5μg/ml^50μg/ml,线性良好,相关系数均>0.999;添加高、中、低3个浓度(1 mg/kg、10 mg/kg、40 mg/kg)作为加标,平均加标回收率为82.1%~110.2%;相对标准偏差(RSD)均<6.5%。检出限(S/N=3)为0.04 mg/kg^0.10 mg/kg,定量限(S/N=10)为0.06 mg/kg^0.30 mg/kg。结论采用自制混合固相萃取柱,样品处理简便、快捷。高效液相色谱法同时检测5种组分,可以保证准确度、灵敏度、精密度。降低时间、人员、环境成本,对于没有液相色谱质谱联用的基层单位,在豆芽检测中,具有一定的推广意义。

Objective To establish a method for the determination of 6 - Benzylaminopurine, isopentennyladenine, gibberellin, 4 -chlorophenoxyacetic acid, 4 -fluorophenoxyaeetie acid in bean sprout using homemade mixed solid phase extraction column coupled with high performance liquid chromatography（HPLC）. Methods The sample was extracted by 20 ml acetonitrile, de- hydrated by salt, then centrifugated, and purified by a homemade mixed solid phase extraction column. Chromatographic analy- sis was carried out on Spursil C18 - EP with aeetonitrile and O. 5% phosphoric acid as mobile phase. The detection wavelength was 209 nm. It was quantified with external standard method. Results Good linearity was obtained when the concentrations of the 5 plant regulators were within 0.5 μg/ml - 50 μg/ml, with the correlation coefficients （r） greater than 0. 999. Three levels of spiked recoveries（ 1 mg/kg, 10 mg/kg, 40 mg/kg） were carried out using blank bean sprout extraction as substrate, the re- coveries were within 82.1% - 110.0%, and the relative standard deviations （RSDs） was less than 6.5 %. The qualitative limits of detection （ S/N = 3 ） were within 0.04 mg/kg -0.1 mg/kg, and the quantitative limits （ S/N = 10） were within 0.06 mg/kg - 0.3 mg/kg. Conclusion The method is simple and easy to operate using homemade mixed solid phase extraction column. Sim- ultaneous determination of five PGRs by high performance liquid chromatography can ensure the corresponding accuracy, sensi- tivity and precision, reducing time, personnel, environmental costs. This method has some significance for the promotion of the basic institutions without LC - MS/MS.