超高效液相色谱-串联质谱法测定食用菌中维生素D_2含量
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摘要
采用超高效液相色谱-串联质谱(ultra performance liquid chromatography-mass spectrometry,UPLCMS/MS)技术建立食用菌中维生素D_2的测定方法;优化质谱条件中的加速电压及碰撞电压,色谱条件中的色谱柱、流动相及其比例。优化结果如下,加速电压选择120 eV,定量子离子与定性子离子的碰撞电压分别选择22 eV和12 eV,色谱柱选择ZORBAX Eclipse plus C18(2.1 mm×150 mm,3.5μm),流动相选择甲醇-水,其比例为95:5(v/v)。方法学考察结果表明,在0.1～40μg/mL浓度范围内,维生素D_2的峰面积与其浓度线性关系良好,相关系数大于0.999,检出限为20.0μg/kg,定量限为60.0μg/kg;回收率为77.0%～92.5%,相对标准偏差(relative stardard deviation,RSD)为2.1%～4.9%,方法准确可靠,可满足食用菌中维生素D_2含量的测定需要。对0.5、1.0、1.5、2.0 kJ/m~2不同剂量短波紫外线照射后的双孢蘑菇(Agaricus bisporus)、糙皮侧耳(Pleurotus ostreatus)和香菇(Lentinus edodes)中维生素D_2的含量进行测定,结果表明4种剂量短波紫外线照射均能显著提高3种食用菌中维生素D_2的含量,在实验范围内,维生素D_2的含量随照射剂量的增加而增加。同一照射剂量处理,糙皮侧耳所含维生素D_2最多,其次为双孢蘑菇、香菇。
Ultra performance liquid chromatography-mass spectrometry( UPLC-MS/MS) was used to determine vitamin D_2 content in edible mushrooms. First, both mass spectrometry and chromatography conditions were optimized. For the mass spectrometry, the optimal fragmentor voltage was 120 eV, and the collision energy of quantitative and qualitative ions were 22 eV and 12 eV, respectively. For the liquid chromatography, ZORBAX Eclipse plus C18(2. 1 mm × 150 mm,3. 5 μm) was used as the analytical column, and the mobile phase was methanol-water with a volumetric ratio of 95:5. There was a good linear relationship between peak area and vitamin D_2 concentration in the range of 0.1 ～40 g/mL. The correlation coefficient of the resultant standard curve was over 0.999. The detection limit was 20.0 μg/kg and the limit of quantity was 60.0 μg/kg. The recovery ratio was 77.0%～92.5%,and the RSD value was 2.4% ～4.9%.The method was accurate and reliable for the quantitative determination of vitamin D_2 in edible mushrooms.Then the method was used to determine the content of vitamin D_2 in white button mushroom,oyster mushroom and shiitake mushroom after different doses of ultraviolet-C irradiation(0.5、1.0、1.5、2.0 kJ/m~2). The results showed that vitamin D_2 in all three edible mushrooms increased significantly after UV irradiation in a dose-dependent manner. Under the same irradiation dose, oyster mushroom contained the highest vitamin D_2, followed by white button mushroom and shiitake mushroom.
Ultra performance liquid chromatography-mass spectrometry( UPLC-MS/MS) was used to determine vitamin D_2 content in edible mushrooms. First, both mass spectrometry and chromatography conditions were optimized. For the mass spectrometry, the optimal fragmentor voltage was 120 eV, and the collision energy of quantitative and qualitative ions were 22 eV and 12 eV, respectively. For the liquid chromatography, ZORBAX Eclipse plus C18(2. 1 mm × 150 mm,3. 5 μm) was used as the analytical column, and the mobile phase was methanol-water with a volumetric ratio of 95:5. There was a good linear relationship between peak area and vitamin D_2 concentration in the range of 0.1 ～40 g/mL. The correlation coefficient of the resultant standard curve was over 0.999. The detection limit was 20.0 μg/kg and the limit of quantity was 60.0 μg/kg. The recovery ratio was 77.0%～92.5%,and the RSD value was 2.4% ～4.9%.The method was accurate and reliable for the quantitative determination of vitamin D_2 in edible mushrooms.Then the method was used to determine the content of vitamin D_2 in white button mushroom,oyster mushroom and shiitake mushroom after different doses of ultraviolet-C irradiation(0.5、1.0、1.5、2.0 kJ/m~2). The results showed that vitamin D_2 in all three edible mushrooms increased significantly after UV irradiation in a dose-dependent manner. Under the same irradiation dose, oyster mushroom contained the highest vitamin D_2, followed by white button mushroom and shiitake mushroom.
引文
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[12] KOYYALAMUDI SR, JEONG SC, PANG G, et al. Concentration of vitamin D2 in white button mushrooms(Agaricus bisporus)exposed to pulsed UV light[J]. Journal of Food Composition and Analysis, 2011, 24(7):976-979.
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[14] KOYYALAMUDI SR, JEONG SC, SONG CH, et al. Vitamin D_2 formation and bioavailability from Agaricus bisporus button mushrooms treated with ultraviolet irradiation[J]. Journal of Agricultural and Food Chemistry,2009, 57(8):3351-3355.
[15] JASINGHE VJ, PERERA CO. Ultraviolet irradiation:The generator of vitamin D_2 in edible mushrooms[J].Food Chemistry, 2006, 95(4):638-643.
[16]黄芳,吴惠勤,杭义萍.高效液相色谱-串联质谱法测定食品中维生素D_2和维生素D_3[J].理化检验(化学分册),2011,47(5):577-579,582.
[17]朱姜,张鹏,杨嘉,等.高效液相色谱-串联质谱法测定奶粉中的维生素A、维生素D、维生素E[J].中国卫生检验杂志,2015, 25(11):1733-1736,1739.
[18] JASINGHE VJ, PERERA CO. Distribution of ergosterol in different tissues of mushrooms and its effect on the conversion of ergosterol to vitamin D2 by UV irradiation[J]. Food Chemistry, 2005, 92(3):541-546.
[19] KO JA, LEE BH, LEE JS, et al. Effect of UV-B exposure on the concentration of vitamin D2 in sliced shiitake mushroom(Lentinus edodes)and white button mushroom(Agaricus bisporus)[J]. Journal of Agricultural and Food Chemistry, 2008, 56(10):3671-3674.
[20] PLUDOWSKI P, HOLICK MF, GRANT WB, et al. Vitamin D supplementation guidelines[J]. Journal of Steroid Biochemistry&Molecular Biology, 2018, 175:125-135.
[2]李金潺,卫丹丹,李小峰,等.维生素D在自身免疫性疾病中的作用[J].中国全科医学,2015, 18(18):2184-2187.
[3] TAOFIQ O, FERNANDES A, BARROS L, et al. UV-irradiated mushrooms as a source of vitamin D_2:A review[J]. Trends in Food Science&Technology, 2017, 70:82-94.
[4] TEICHMANNA, DUTTA PC, STAFF AS A, et al. Sterol and vitamin D_2 concentrations in cultivated and wild grown mushrooms:Effects of UV irradiation[J]. LWT-Food Science and Technology, 2007, 40(5):815-822.
[5] MATTILA P, LAMPI AM, RONKAINEN R, et al. Sterol and vitamin D_2 contents in some wild and cultivated mushrooms[J]. Food Chemistry, 2002, 76(3):293-298.
[6]张金霞,陈强,黄晨阳,等.食用菌产业发展历史、现状与趋势[J].菌物学报,2015, 34(4):524-540.
[7]孙梦姣.紫外照射提高双孢蘑菇中维生素D2含量的研究[D].杭州:浙江大学,2013.
[8] GUAN WQ, ZHANG J, YAN RX, et al. Effects of UV-C treatment and cold storage on ergosterol and vitamin D2 contents in different parts of white and brown mushroom(Agaricus bisporus)[J]. Food Chemistry, 2016, 210:129-134.
[9]张焱,程学新,刘颙颙,等.光化学合成维生素D[J].精细与专用化学品,2005, 13(5):5-7.
[10] KEEGAN RH, LU ZR, BOGUSZ JM, et al. Photobiology of vitamin D in mushrooms and its bioavailability in humans[J]. Dermato-Endocrinology, 2013, 5(1):165-176.
[11]邓卿丰.高效液相色谱-质谱技术在食品安全分析中的应用进展[J].现代食品,2018(20):85-87.
[12] KOYYALAMUDI SR, JEONG SC, PANG G, et al. Concentration of vitamin D2 in white button mushrooms(Agaricus bisporus)exposed to pulsed UV light[J]. Journal of Food Composition and Analysis, 2011, 24(7):976-979.
[13] SLAWINSKA A, FORNAL E, RADZKI W, et al. Study on vitamin D_2 stability in dried mushrooms during drying and storage[J]. Food Chemistry, 2016, 199:203-209.
[14] KOYYALAMUDI SR, JEONG SC, SONG CH, et al. Vitamin D_2 formation and bioavailability from Agaricus bisporus button mushrooms treated with ultraviolet irradiation[J]. Journal of Agricultural and Food Chemistry,2009, 57(8):3351-3355.
[15] JASINGHE VJ, PERERA CO. Ultraviolet irradiation:The generator of vitamin D_2 in edible mushrooms[J].Food Chemistry, 2006, 95(4):638-643.
[16]黄芳,吴惠勤,杭义萍.高效液相色谱-串联质谱法测定食品中维生素D_2和维生素D_3[J].理化检验(化学分册),2011,47(5):577-579,582.
[17]朱姜,张鹏,杨嘉,等.高效液相色谱-串联质谱法测定奶粉中的维生素A、维生素D、维生素E[J].中国卫生检验杂志,2015, 25(11):1733-1736,1739.
[18] JASINGHE VJ, PERERA CO. Distribution of ergosterol in different tissues of mushrooms and its effect on the conversion of ergosterol to vitamin D2 by UV irradiation[J]. Food Chemistry, 2005, 92(3):541-546.
[19] KO JA, LEE BH, LEE JS, et al. Effect of UV-B exposure on the concentration of vitamin D2 in sliced shiitake mushroom(Lentinus edodes)and white button mushroom(Agaricus bisporus)[J]. Journal of Agricultural and Food Chemistry, 2008, 56(10):3671-3674.
[20] PLUDOWSKI P, HOLICK MF, GRANT WB, et al. Vitamin D supplementation guidelines[J]. Journal of Steroid Biochemistry&Molecular Biology, 2018, 175:125-135.