该文建立液相色谱-稳定同位素比值质谱联用(liquid chromatography-stable isotope ratio mass spectrometry,LC-IRMS)检测抗坏血酸δ13C值的分析方法,用于鉴别针叶樱桃粉中抗坏血酸天然来源的真实性。样品中抗坏血酸经液相色谱在线分离纯化,优化后色谱条件为:Syncronis C18(250 mm×4.6 mm, 5 μm)色谱柱,流动相为水-pH 2的硫酸溶液(90∶10, 体积比),流速0.250 mL/min,色谱柱温度30 ℃,进样量10 μL,通过 LC-IsoLink 实现目标物全部氧化为CO2气体,最终以气态形式进入稳定同位素质谱仪,直接检测样品中抗坏血酸的δ13C,该方法结果稳定、准确。分别测定了7个合成来源的维生素C片和19个针叶樱桃粉,结果表明,天然来源抗坏血酸δ13C值为-25.00‰~-22.01‰,合成来源抗坏血酸δ13C值为-11.74‰~-10.28‰,两者分布显著性差异,该方法可用于抗坏血酸产品标识的真实性鉴别研究。
An analytical method for the detection of ascorbic acid δ13C value by liquid chromatography-stable isotope ratio mass spectrometry (LC-IRMS) was established to identify the authenticity of the natural source of ascorbic acid in acerola cherry powder. Ascorbic acid in the sample was separated and purified by liquid chromatography, and the optimized chromatographic conditions were that Syncronis C18 (250 mm×4.6 mm, 5 mm) column was used, the mobile phase was the aqueous-sulfuric acid solution (90∶10, volume ratio), the flow rate was 0.250 mL/min, the column temperature was 30 ℃, the injection volume was 10 mL, and all the targets were oxidized to CO2 gas by LC-IsoLink, and finally entered the stable isotope mass spectrometer in gaseous form. The δ13C of ascorbic acid in the sample was directly detected, and the results of this method were stable and accurate. Results showed that the distribution range of natural ascorbic acid δ13C value was -25.00‰--22.01‰, and the distribution of synthetic ascorbic acid δ13C value was -11.74‰--10.28‰, which was significantly different. This method could be used for the authenticity identification of ascorbic acid product identification.
[1] NJUS D, KELLEY P M, TU Y J, et al.Ascorbic acid:The chemistry underlying its antioxidant properties[J].Free Radical Biology and Medicine, 2020, 159:37-43.
[2] ZOU M Y, NIE S P, YIN J Y, et al.Ascorbic acid induced degradation of polysaccharide from natural products:A review[J].International Journal of Biological Macromolecules, 2020, 151:483-491.
[3] KABASAKALIS V,SIOPIDOU D, MOSHATOU E.Ascorbic acid content of commercial fruit juices and its rate of loss upon storage[J].Food Chemistry, 2000, 70(3):325-328.
[4] CAMIN F, BONTEMPO L, PERINI M, et al.Stable isotope ratio analysis for assessing the authenticity of food of animal origin[J].Comprehensive Reviews in Food Science and Food Safety, 2016, 15(5):868-877.
[5] AKAMATSU F, SHIMIZU H, HAYASHI S, et al.Chemometric approaches for determining the geographical origin of Japanese Chardonnay wines using oxygen stable isotope and multi-element analyses[J].Food Chemistry, 2022, 371(1):131113.
[6] 王道兵, 钟其顶, 高红波, 等.GasBenchⅡ-IRMS测定葡萄酒水中δ18O方法研究[J].质谱学报, 2014, 35(4):355-360.
WANG D B, ZHONG Q D, GAO H B, et al.Determination of oxygen isotope ratios of water in wine by GasBenchⅡ-IRMS[J].Journal of Chinese Mass Spectrometry Society, 2014, 35(4):355-360.
[7] PERINI M, PAOLINI M, SIMONI M, et al.Stable isotope ratio analysis for verifying the authenticity of balsamic and wine vinegar[J].Journal of Agricultural and Food Chemistry, 2014, 62(32):8197-8203.
[8] KOBAYASHI K, TANAKA M, TANABE S, et al.Distinguishing glutamic acid in foodstuffs and monosodium glutamate used as seasoning by stable carbon and nitrogen isotope ratios[J].Heliyon, 2018, 4(9):e00800.
[9] GREULE M, MOSANDL A, HAMILTON J T G, et al.Comment on authenticity and traceability of Vanilla flavors by analysis of stable isotopes of carbon and hydrogen[J].Journal of Agricultural and Food Chemistry, 2015, 63(21):5305-5306.
[10] 国辉, 班楠, 王道兵, 等.酱油中内源性苯甲酸本底含量调查与溯源分析[J].食品科学, 2021, 42(20):286-291.
LI G H, BAN N, WANG D B, et al.Investigation and traceability analysis of endogenous benzoic acid in soy sauce[J].Food Science, 2021, 42(20):286-291.
[11] ELGADI S, OUHAMMOU A, TAOUS F, et al.Combination of stable isotopes and fatty acid composition for geographical origin discrimination of one argan oil vintage[J].Foods, 2021, 10(6):1274.
[12] JIMÉNEZ-MORILLO N T, PALMA V, GARCIA R, et al.Multivariate geostatistical analysis of stable isotopes in Portuguese varietal extra virgin olive oils[J].Microchemical Journal, 2020, 157:105044.
[13] 王道兵, 岳红卫, 高冠勇, 等.基于稳定氢氧同位素技术的花生油掺假检测技术研究[J].中国粮油学报, 2021, 36(8):118-122.
WANG D B, YUE H W, GAO G Y, et al.Research on detection of adulterated peanut oil by stable hydrogen/oxygen isotope analysis[J].Journal of the Chinese Cereals and Oils Association, 2021, 36(8):118-122.
[14] 马玉华, 唐方东, 刘佳煜, 等.稳定同位素比技术用于橄榄油的掺假鉴定[J].质谱学报, 2021, 42(2):189-196.
MA Y H, TANG F D, LIU J Y, et al.Olive oil adulteration identification using stable isotope ratio technique[J].Journal of Chinese Mass Spectrometry Society, 2021, 42(2):189-196.
[15] MAGDAS D A, GUYON F, PUSCAS R, et al.Applications of emerging stable isotopes and elemental markers for geographical and varietal recognition of Romanian and French honeys[J].Food Chemistry, 2021, 334:127599.
[16] SCHIMMELMANN A, QI H P, DUNN P J H, et al.Food matrix reference materials for hydrogen, carbon, nitrogen, oxygen, and sulfur stable isotope-ratio measurements:Collagens, flours, honeys, and vegetable oils[J].Journal of Agricultural and Food Chemistry, 2020, 68(39):10852-10864.
[17] 黄学者, 王蕾, 魏红跃, 等.云南3种特色蜂蜜糖组分的稳定碳同位素比值测定与分析[J].核农学报, 2020, 34(S1):79-88.
HUANG X Z, WANG L, WEI H Y, et al.Determination and analysis of stable δ13C value of sugar components in three kinds of characteristic honey from Yunnan Province[J].Journal of Nuclear Agricultural Sciences, 2020, 34(S1):79-88.
[18] HANAMURA T, UCHIDA E, ISHIGURO Y, et al.Discrimination between ascorbic acid from acerola and of synthetic origin by measuring 13C and 18O stable isotope ratios[J].Chemistry Letters, 2012, 41(9):929-931.
[19] ALBERTINO A, BARGE A, CRAVOTTO G, et al.Natural origin of ascorbic acid:Validation by 13C NMR and IRMS[J].Food Chemistry, 2009, 112(3):715-720.
[20] 李学民, 贾光群, 曹彦忠, 等.液相色谱-同位素比质谱法同时测定葡萄酒中甘油和乙醇的δ13C值[J].色谱, 2013, 31(12):1201-1205.
LI X M, JIA G Q, CAO Y Z, et al.Simultaneous determination of δ13C values of glycerol and ethanol in wine by liquid chromatography coupled with isotope ratio mass spectrometry[J].Chinese Journal of Chromatography, 2013, 31(12):1201-1205.
