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超高效液相色谱-串联质谱法同时分析紫薯中15种酚酸类组分

刘文静 黄彪 林香信 方灵 吴建鸿 李巍 吴妙鸿

刘文静,黄彪,林香信,等. 超高效液相色谱-串联质谱法同时分析紫薯中15种酚酸类组分 [J]. 福建农业学报,2019,34(10):1203−1210. doi: 10.19303/j.issn.1008-0384.2019.10.014
引用本文: 刘文静,黄彪,林香信,等. 超高效液相色谱-串联质谱法同时分析紫薯中15种酚酸类组分 [J]. 福建农业学报,2019,34(10):1203−1210. doi: 10.19303/j.issn.1008-0384.2019.10.014
LIU W J, HUANG B, LIN X X, et al. Simultaneous Determination of 15 Phenolic Compounds in Purple Potato by High Performance Liquid Chromatography-tandem Mass Spectrometry [J]. Fujian Journal of Agricultural Sciences,2019,34(10):1203−1210. doi: 10.19303/j.issn.1008-0384.2019.10.014
Citation: LIU W J, HUANG B, LIN X X, et al. Simultaneous Determination of 15 Phenolic Compounds in Purple Potato by High Performance Liquid Chromatography-tandem Mass Spectrometry [J]. Fujian Journal of Agricultural Sciences,2019,34(10):1203−1210. doi: 10.19303/j.issn.1008-0384.2019.10.014

超高效液相色谱-串联质谱法同时分析紫薯中15种酚酸类组分

doi: 10.19303/j.issn.1008-0384.2019.10.014
基金项目: 福建省科技计划公益类专项(2017R1018-2);福建省农业科学院青年人才创新项目(YC2018-11)
详细信息
    作者简介:

    刘文静(1982−),女,硕士,助理研究员,主要从事农产品质量安全研究(E-mail:411935637@qq.com)

    通讯作者:

    林香信(1972−),男,副研究员,主要从事农产品质量安全研究(E-mail:1353645173@qq.com

  • 中图分类号: TS 215

Simultaneous Determination of 15 Phenolic Compounds in Purple Potato by High Performance Liquid Chromatography-tandem Mass Spectrometry

  • 摘要:   目的  运用超高效液相色谱-串联质谱(UPLC-MS/MS)分析不同品种紫薯中酚酸类组分及含量,为紫薯中酚酸类组分的提取、分析提供理论依据。  方法  建立超高效液相色谱-串联质谱法同时检测紫薯中15种酚酸类组分的方法,分析8种不同品种紫薯中酚酸类组分的含量。样品经甲醇-水溶液超声提取后,使用Waters T3 C18色谱柱(250 mm × 4.6 mm,5 μm),以0.1%(v/v)甲酸-5 mmol·L−1乙酸铵水溶液和乙腈为流动相进行梯度洗脱,负离子扫描方式下多反应监测(MRM)模式测定。  结果  在优化的试验条件下,15种酚酸类组分在相应的浓度范围内具有良好的线性关系,相关系数(r)为0.995 6~0.999 7,方法检出限和定量限分别为0.3~5.8 μg·L−1(S/N ≥3)和1.0~15.2 μg·L−1(S/N ≥10);加标试验平均回收率73.3%~97.1%,相对标准偏差1.8%~8.9%。  结论  在所分析的紫薯样品中共检出5种酚酸类组分,含量0.102~1 698.490 mg·kg−1,其中绿原酸含量最高,对香豆酸含量最低。该法灵敏、快速、准确,适用于紫薯中酚酸类组分的定性定量分析。
  • 表  1  酚酸类组分检测的质谱参数条件

    Table  1.   Parameters applied for phenolic determination by LC-MS/MS

    组分名称
    Compound
    保留时间
    Retention time /min
    母离子
    Parents ion/ (m·z−1)
    子离子
    Daughter ion/ (m·z−1)
    锥孔电压
    Cone voltage /V
    碰撞能量
    Collision energy/eV
    3,4,5-三羟基苯甲酸 3,4,5-Trihydroxybenzoic acid 1.68 168.8 125.0*, 79.0 −36 14, 20
    3,5-二羟基苯甲酸 3,5-Dihydroxybenzoic acid 2.62 152.9 108.8*, 80.9 −40 14, 18
    绿原酸 Chlorogenic acid 3.25 353.2 191.0*, 92.9 −30 16, 38
    对羟基苯甲酸 p−Hydroxybenzoic acid 3.72 136.9 92.9*, 137.4 −36 12, 16
    2,5-二羟基苯甲酸 2,5-Dihydroxybenzoic acid 3.82 152.9 108.8*, 80.9 −40 14, 18
    香草酸 Vanillic acid 3.85 167.0 151.9*, 108.0 −36 14, 18
    咖啡酸 Caffeic acid 3.88 179.1 134.9*, 89.0 −30 16, 32
    丁香酸 Syringate 3.97 197.0 181.9*, 123.0 −48 14, 26
    对香豆酸 Coumalic acid 4.33 163.0 119.0*, 93.3 −44 16, 26
    异绿原酸 A Isochlorogenic acid A 4.39 514.9 352.9*, 190.9 −30 16, 32
    芥子酸 Sinapic acid 4.55 223.0 208.0*, 164.0 −44 16, 14
    阿魏酸 Ferulic acid 4.59 193.0 133.9*, 177.9 −38 16, 14
    3-羟基肉桂酸 3-Hydrocycinnamic acid 4.64 163.0 119.0*, 93.3 −44 16, 26
    2-羟基肉桂酸 2-Hydrocycinnamic acid 4.83 163.0 119.0*, 93.3 −44 16, 26
    水杨酸 Salicylic acid 5.18 136.9 92.9*, 137.4 −36 16, 12
    注:*为定量离子。
    Note:* means quantitative ions.
    下载: 导出CSV

    表  2  提取溶液对酚类物质提取效果比较

    Table  2.   Comparison of the effect of extraction solution

    提取溶液
    Extraction solution
    吸光度值
    absorbance value
    多酚含量
    Content/(mg·g−1)
    30%甲醇水溶液 Methanol0.5493.14
    50%甲醇水溶液 Methanol0.6913.95
    70%甲醇水溶液 Methanol0.7364.20
    下载: 导出CSV

    表  3  15种酚酸类组分检测的方法学验证

    Table  3.   Methodology validation of UPLC-MS/MS

    组分名称
    Compound
    线性范围
    Line arrange/
    (µg·L−1
    线性方程
    Linear equation
    相关系数r检出限
    Limits of detection/
    (µg·L−1)
    定量限
    Limits of quantitation/
    (µg·L−1)
    3,4,5-三羟基苯甲酸 3,4,5-Trihydroxybenzoic acid 20~8 000 y = 89.9x +3 841.9 0.998 4 3.2 10.2
    3,5-二羟基苯甲酸 3,5-Dihydroxybenzoic acid 20~8 000 y = 456.2x + 22 407.5 0.999 2 1.8 5.0
    绿原酸 Chlorogenic acid 20~8 000 y = 311.5x + 99.8 0.999 5 2.1 5.0
    对羟基苯甲酸 p-Hydroxybenzoic acid 10~4 000 y = 819.2x + 1 232.2 0.997 1 1.2 3.2
    2,5-二羟基苯甲酸 2,5-Dihydroxybenzoic acid 10~4 000 y = 857.4x + 805.4 0.997 6 1.2 3.5
    香草酸 Vanillic acid 10~4 000 y = 1 875.0x + 9 070.7 0.995 6 0.5 2.0
    咖啡酸 Caffeic acid 50~20 000 y = 80.7x + 18.2 0.999 0 3.6 10.0
    丁香酸 Syringate 50~20 000 y = 35.2x + 1 100.0 0.998 4 5.8 15.2
    对香豆酸 Coumalic acid 10~4 000 y = 1 452.9x + 734.0 0.998 7 0.4 1.5
    异绿原酸 A Isochlorogenic acid A 20~8 000 y= 368.1x – 90.0 0.999 0 2.2 6.8
    芥子酸 Sinapic acid 20~8 000 y = 340.5x + 214.8 0.999 4 2.0 6.0
    阿魏酸 Ferulic acid 20~8 000 y = 607.7x + 28.4 0.999 7 1.5 4.5
    3-羟基肉桂酸 3-Hydrocycinnamic acid 10~4 000 y = 1 180.1x + 523.9 0.998 9 0.6 2.0
    2-羟基肉桂酸 2-Hydrocycinnamic acid 10~4 000 y = 1 211.5x + 684.7 0.999 1 0.6 2.0
    水杨酸 Salicylic acid 10~4 000 y = 2 493.6x + 1 857.8 0.997 6 0.3 1.0
    下载: 导出CSV

    表  4  紫薯中15种酚类化合物的加标回收试验(n=6)

    Table  4.   Recovery rates of 15 phenols in actual samples

    组分名称
    Compound
    本底值
    Background/ (mg·kg−1)
    加标量
    Addition /(mg·kg−1)
    测量值
    Measured value /(mg·kg−1)
    回收率
    Recovery /%
    标准偏差
    Standard deviation /%
    3,4,5-三羟基苯甲酸
    3,4,5-Trihydroxybenzoic acid
    ND 0.400 0.317 79.2 4.7
    0.800 0.586 73.3 1.7
    2.000 1.942 97.1 3.1
    3,5-二羟基苯甲酸
    3,5-Dihydroxybenzoic acid
    ND 0.400 0.348 87.0 6.6
    0.800 0.706 88.2 3.1
    2.000 1.854 92.7 5.5
    绿原酸
    Chlorogenic acid
    498.161 10.00 506.931 87.7 5.4
    20.00 513.121 74.8 4.9
    50.00 536.761 77.2 6.3
    对羟基苯甲酸
    p-Hydroxybenzoic acid
    ND 0.200 0.175 87.7 7.0
    0.400 0.343 85.7 3.2
    1.000 0.868 86.8 2.1
    2,5-二羟基苯甲酸
    2,5-Dihydroxybenzoic acid
    ND 0.200 0.154 77.2 4.2
    0.400 0.324 81.1 4.3
    1.000 0.904 90.4 2.7
    香草酸
    Vanillic acid
    9.152 1.000 1.7702 85.5 3.6
    2.000 2.6052 84.5 7.9
    5.000 5.3602 88.9 3.2
    咖啡酸
    Caffeic acid
    ND 1.000 0.783 78.3 5.5
    2.000 1.588 79.4 4.3
    5.000 4.055 81.1 5.9
    丁香酸
    Syringate
    1.384 1.000 0.9774 83.9 6.9
    2.000 2.0304 94.6 8.7
    5.000 4.6534 90.3 8.9
    对香豆酸
    Coumalic acid
    1.017 0.200 1.162 72.3 7.7
    0.400 1.322 76.3 1.9
    1.000 1.791 77.4 8.5
    异绿原酸A
    Isochlorogenic acid A
    118.074 10.00 126.644 85.7 5.1
    20.00 135.334 86.3 6.2
    50.00 163.174 90.2 3.8
    芥子酸
    Sinapic acid
    ND 0.400 0.304 76.1 5.6
    0.800 0.603 75.4 3.7
    2.000 1.772 88.6 4.8
    阿魏酸
    Ferulic acid
    ND 0.200 0.150 74.9 5.0
    0.400 0.324 80.9 7.1
    1.000 0.913 91.3 6.2
    3-羟基肉桂酸
    3-Hydrocycinnamic acid
    ND 0.200 0.149 74.5 6.0
    0.400 0.325 81.3 2.0
    1.000 0.894 89.4 7.6
    2-羟基肉桂酸
    2-Hydrocycinnamic acid
    ND 0.200 0.158 79.2 4.2
    0.400 0.351 87.8 4.3
    1.000 0.839 83.9 2.7
    水杨酸
    Salicylic acid
    ND 0.200 0.175 87.3 4.6
    0.400 0.323 80.7 3.9
    1.000 0.968 96.8 2.2
    注:ND:未检出。
    Note: ND means not detected.
    下载: 导出CSV

    表  5  不同紫薯酚酸类组分含量

    Table  5.   Contents of phenols in purple potato samples (单位:mg·kg−1,n=3)

    样品 Sample香草酸 Vanillic acid咖啡酸 Caffeic acid对香豆酸Coumalic acid绿原酸 Chlorogenic acid异绿原酸 A Isochlorogenic acid A
    福薯9号 Ganshu 9 1.241 3.293 0.137 291.439 34.099
    莆紫18号 Puzi 18 1.700 9.819 0.112 1 463.459 112.602
    福薯24号 Fushu 24 1.613 3.386 1.173 541.270 58.864
    龙紫6号 Longzi 6 1.474 3.806 0.381 1 158.459 135.749
    莆紫3号 Puzi 3 1.896 5.409 0.142 644.501 46.524
    龙紫4号 Longzi 4 1.835 4.071 0.102 1 250.894 112.611
    福宁紫4号 Funingzi 4 1.553 7.869 0.113 469.991 96.853
    福宁紫3号 Funingzi 3 2.237 10.722 0.122 1 698.490 202.682
    下载: 导出CSV
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出版历程
  • 收稿日期:  2019-08-16
  • 修回日期:  2019-10-08
  • 刊出日期:  2019-10-01

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