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目的:建立超高效液相色谱质谱分析方法,比较不同蜜源蜂蜜中黄酮类组分的区别。方法:不同蜜源蜂蜜经D_(101)型大孔吸附树脂提取纯化后采用UPLC-MS/MS进行分析。色谱柱采用Inertsil ODS-3柱(2.1 mm×75 mm,2μm),以甲醇-0.02%甲酸水溶液为流动相梯度洗脱,流速0.4 m L·min~(-1),柱温40℃;以儿茶素、表儿茶素、芦丁、芸香柚皮苷、桑黄素、杨梅素、山柰酚、生松素等16个黄酮类对照品作为对照,采用电喷雾离子源(ESI),多反应监测(MRM)进行分析。结果:儿茶素、表儿茶素、芸香柚皮苷、芦丁、桑黄素、杨梅素、柚皮素、槲皮素、染料木素、木犀草素、山柰酚、芹菜素、生松素、汉黄芩素、白杨素和高良姜素的质量浓度分别在0.835~83.5μg·m L~(-1)(r=0.998 5)、0.562~22.48μg·m L~(-1)(r=0.999 0)、0.095 2~4.76μg·m L~(-1)(r=0.995 5)、0.052 2~1.307μg·m L~(-1)(r=0.998 0)、1.608~40.2μg·m L~(-1)(r=0.997 0)、0.45~9.0μg·m L~(-1)(r=0.998 5)、0.006 9~0.69μg·m L~(-1)(r=0.999 5)、0.022 5~4.498μg·m L~(-1)(r=0.999 5)、0.040 6~4.055μg·m L~(-1)(r=0.999 5)、0.073 3~3.665μg·m L~(-1)(r=0.999 5)、3.26~32.6μg·m L~(-1)(r=0.996 5)、0.021 4~1.072μg·m L~(-1)(r=0.999 0)、0.047 2~1.18μg·m L~(-1)(r=0.998 5)、0.001 4~0.353μg·m L~(-1)(r=0.999 5)、0.058~2.925μg·m L~(-1)(r=0.999 5)、0.222 4~11.12μg·m L~(-1)(r=0.997 5)范围内与峰面积呈良好的线性关系,提取回收率分别为2.7%、3.8%、0.26%、0.00%、70.6%、59.3%、83.0%、87.8%、83.4%、81.1%、93.3%、89.9%、86.1%、95.9%、89.8和93.3%,基质效应(n=5)在94.29%~115.72%之间,18 h内基本稳定,其中杨梅素6 h内稳定(RSD=2.3%)。不同蜜源蜂蜜之间黄酮类组分及其含量有一定程度的差异。如洋槐蜜中槲皮素、白杨素、生松素和山柰酚为主,木犀草素、染料木素、汉黄芩素等为辅;柑橘蜜中以槲皮素、山柰酚、高良姜素和汉黄芩素为主,木犀草素和芹菜素为辅;油菜蜜中以山柰酚和槲皮素为主,白杨素、生松素、柚皮素、染料木素、木犀草素和汉黄芩素等为辅等。某些化合物有可能成为蜂蜜中的潜在标志组分,如洋槐蜜中的染料木素及柑橘蜜中的汉黄芩素,与其他蜜源蜂蜜中的组分含量相比,具有含量占比高或蜂蜜和蜜源花中均有等特点。结论:该方法选择性强,灵敏度高,所得结果准确可靠,同时有助于单花蜜蜜源来源的鉴定。
OBJECTIVE: To establish a method for the determination of flavonoids in honey of different nectar sources by ultra performance liquid chromatography-mass spectrometry. Methods: The honey of different nectar sources was purified by D_ (101) macroporous adsorption resin and analyzed by UPLC-MS / MS. The column was eluted with a mobile phase of methanol-0.02% formic acid at a flow rate of 0.4 mL · min -1 on an Inertsil ODS-3 column (2.1 mm × 75 mm, 2 μm) Sixteen flavonoid reference materials, catechin, epicatechin, rutin, rutin, naringin, myricetin, myricetin and kaempferol, were used as control. Electrospray ionization (ESI) Multiple Reaction Monitoring (MRM) was analyzed. Results: Catechin, epicatechin, rutin naringin, rutin, anthracene, myricetin, naringenin, quercetin, genistein, luteolin, kaempferol, apigenin, The mass concentrations of spironolactone, wogonin, chrysin, and galangin were between 0.835 and 83.5 μg · m L -1 (r = 0.998 5) and 0.562 ~ 22.48 μg · m L -1 ( r = 0.999 0), 0.095 2 ~ 4.76μg · m L -1 (r = 0.995 5), 0.052 2 ~ 1.307μg · m L -1 (r = 0.998 0), 1.608 ~ 40.2μg · M L -1 (r = 0.997 0), 0.45-9.0 μg · m L -1 (r = 0.998 5), 0.006 9-0.69 μg · m L -1 (r = 0.999 5), 0.022 5 ~ 4.498μg · m L -1 (r = 0.999 5), 0.040 6 ~ 4.055μg · m L -1 (r = 0.999 5), 0.073 3 ~ 3.665μg · (r = 0.999 5), 3.26 ~ 32.6 μg · m L -1 (r = 0.996 5), 0.021 4 ~ 1.072 μg · m L -1 (r = 0.999 (R = 0.995 5), 0.001 4 ~ 0.353 μg · m L -1 (r = 0.999 5), 0.058 ~ 2.925 μg · m L ~ (-1) (r = 0.999 5), 0.222 4 ~ 11.12 μg · m L -1 (r = 0.997 5), and the peak area was linear with the recoveries of 2.7% 3.8%, 0.26%, 0.00%, 70.6%, 59.3%, 83.0%, 87.8%, 83.4%, 81.1% 93.3%, 89.9%, 86.1%, 95.9%, 89.8% and 93.3% respectively. The matrix effect (n = 5) was between 94.29% and 115.72% within 18 h. %). Different nectar honey flavonoids and their components have a certain degree of difference. Such as acacia honey quercetin, chrysin, Shengsong Su and kaempferol-based, luteolin, genistein, wogonin and other supplemented by citrus honey quercetin, kaempferol, galangal Succulent baicalin and baicalin, supplemented by luteolin and apigenin; rape honey to kaempferol and quercetin, chrysin, spongin, naringenin, genistein, luteolin and Wogonin and other supplements and so on. Some of the compounds may be potential markers in honey, such as genistein in the acacia honey and wogonin in citrus honey, which have a higher content of honey or higher proportion of honey than honey And nectar flowers have the same characteristics. Conclusion: The method is highly selective and sensitive, and the results are accurate and reliable. It is also helpful to identify the source of honey nectarine.