超高效液相色谱-串联质谱法测定液态乳中11种营养成分
Determination of 11 nutrients in liquid milk by ultra-performance liquid chromatography-tandem mass spectrometry
王雪楠 1王兴龙 2刘少颖 2任一平 3任韧 2张蓉 1张雪松 1王竹4
作者信息
- 1. 中国疾病预防控制中心营养与健康所,北京 100050
- 2. 浙江省杭州市疾病预防控制中心,杭州 310021
- 3. 浙江省疾病预防控制中心,杭州 310057
- 4. 中国疾病预防控制中心营养与健康所,北京 100050;国家卫生健康委公共营养与健康重点实验室,北京 100050
- 折叠
摘要
目的 建立可同时检测液态乳中11种营养成分(硫胺素、核黄素、烟酰胺、烟酸、泛酸、吡哆醇、吡哆醛、吡哆胺、生物素、胆碱、左旋肉碱)的超高效液相色谱-串联质谱法.方法 液态乳样品采用20 mmol/L甲酸铵溶液震荡并于100 ℃水浴加热30 min后,经木瓜蛋白酶和酸性磷酸酶45℃孵育16 h,离心取下层液体进行测定.色谱柱为 ACQUITYTM HSS T3(3.0 mm× 150 mm,1.8 μm),以 2 mmol/L 甲酸铵 0.1％甲酸水溶液和0.1％甲酸乙腈为流动相,采用内标法定量检测.结果 11种营养成分在12 min内可以有效分离,线性相关系数(R2)均大于0.995,方法检出限为0.05～0.50 μg/L,定量限为0.20～1.25 μg/L,三水平加标回收率为85.6％～119.3％,精密度为3.68％～7.82％(n=6).对60份来自5种不同动物的液态乳进行检测,发现不同乳源液态乳中 11种营养成分的含量差异较大,但均未能检出吡哆醇.结论 本方法通过有效酶解可定量检测包括游离态和结合态的11种水溶性营养成分,方法灵敏,重现性良好,精密度和准确度都能满足定量检测需要.
Abstract
OBJECTIVE To establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for simultaneous determination of 11 nutritional components(thiamine,riboflavin,nicotinamide,nicotinic acid,pantothenic acid,pyridoxine,pyridoxal,pyridoxamine,biotin,choline,L-carnitine)in liquid milk.METHODS Milk samples were shaken with 20 mmol/L ammonium formate solution and heated in a water bath at 100 ℃ for 30 min,then incubated with papain and acid phosphatase at 45 ℃ for 16 h,the lower liquid was collected after centrifugation for analysis.UPLC separation was performed on an ACQUITYTM HSS T3(3.0 mm×150 mm,1.8 μm)column,2 mmol/L ammonium formate(containing 0.1％formic acid)solution and acetonitrile(containing 0.1％formic acid)were used as mobile phase.Quantitative detection was performed by internal standard method.RESULTS 11 nutritional components can be effectively separated and detected in 12 min,and the linear correlation coefficients(R2)were all above 0.995.The limits of detection(LODs)were between 0.05 and 0.50 μg/L,and the limits of quantification(LOQs)were between 0.20 and 1.25 µg/L.The recovery rates of three-level addition were 85.6％-119.3％,and the precision RSDs were between 3.68％and 7.82％(n=6).Based on the detection of 60 liquid milk samples from 5 different animals,it was found that the contents of 11 nutrients in liquid milk from different milk sources were significantly different,but pyridoxine could not be detected.CONCLUSION The method can quantitatively detect 11 water-soluble nutrients,including free and bound forms,by effective enzymolysis.It is sensitive,reproducible and can meet the needs of quantitative detection.
关键词
B族维生素/左旋肉碱/超高效液相色谱-串联质谱法/液态乳Key words
B vitamins/L-carnitine/ultra-performance liquid chromatography-tandem mass spectrometry/liquid milk引用本文复制引用
基金项目
达能营养中心膳食营养研究与宣教基金(DIC2021-08)
中国营养学会科研基金-海普诺凯母婴营养科研专项(CNS-HPNK2021-67)
杭州市医药卫生科技项目(A20230034)
出版年
2024
NETL
NSTL
万方数据