CN113237964A - 一种发酵蜂花粉特殊成分检测方法 - Google Patents
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- 229940038481 bee pollen Drugs 0.000 title claims abstract description 27
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000001514 detection method Methods 0.000 claims abstract description 17
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 claims abstract description 11
- 239000007788 liquid Substances 0.000 claims abstract description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 54
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 claims description 28
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 claims description 14
- 235000019253 formic acid Nutrition 0.000 claims description 14
- 239000007864 aqueous solution Substances 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
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- 238000001819 mass spectrum Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 4
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 238000000889 atomisation Methods 0.000 claims description 2
- 239000008367 deionised water Substances 0.000 claims description 2
- 229910021641 deionized water Inorganic materials 0.000 claims description 2
- 238000010828 elution Methods 0.000 claims description 2
- 238000004811 liquid chromatography Methods 0.000 claims description 2
- 239000012071 phase Substances 0.000 claims description 2
- 238000012546 transfer Methods 0.000 claims description 2
- 238000005303 weighing Methods 0.000 claims description 2
- 238000002386 leaching Methods 0.000 claims 2
- KSEBMYQBYZTDHS-HWKANZROSA-N ferulic acid Chemical class COC1=CC(\C=C\C(O)=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-N 0.000 abstract description 9
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- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 0.000 description 3
- 229940114124 ferulic acid Drugs 0.000 description 3
- KSEBMYQBYZTDHS-UHFFFAOYSA-N ferulic acid Natural products COC1=CC(C=CC(O)=O)=CC=C1O KSEBMYQBYZTDHS-UHFFFAOYSA-N 0.000 description 3
- 235000001785 ferulic acid Nutrition 0.000 description 3
- 150000002500 ions Chemical class 0.000 description 3
- 238000012827 research and development Methods 0.000 description 3
- QURCVMIEKCOAJU-UHFFFAOYSA-N trans-isoferulic acid Natural products COC1=CC=C(C=CC(O)=O)C=C1O QURCVMIEKCOAJU-UHFFFAOYSA-N 0.000 description 3
- NEAQRZUHTPSBBM-UHFFFAOYSA-N 2-hydroxy-3,3-dimethyl-7-nitro-4h-isoquinolin-1-one Chemical compound C1=C([N+]([O-])=O)C=C2C(=O)N(O)C(C)(C)CC2=C1 NEAQRZUHTPSBBM-UHFFFAOYSA-N 0.000 description 2
- 240000001548 Camellia japonica Species 0.000 description 2
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- 238000012356 Product development Methods 0.000 description 2
- 235000018597 common camellia Nutrition 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- IXORZMNAPKEEDV-UHFFFAOYSA-N gibberellic acid GA3 Natural products OC(=O)C1C2(C3)CC(=C)C3(O)CCC2C2(C=CC3O)C1C3(C)C(=O)O2 IXORZMNAPKEEDV-UHFFFAOYSA-N 0.000 description 2
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- 235000014698 Brassica juncea var multisecta Nutrition 0.000 description 1
- 235000006008 Brassica napus var napus Nutrition 0.000 description 1
- 240000000385 Brassica napus var. napus Species 0.000 description 1
- 235000006618 Brassica rapa subsp oleifera Nutrition 0.000 description 1
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Abstract
一种发酵蜂花粉特殊成分检测方法,包括如下步骤:步骤一:配置检测液;步骤二:将待检测液注入安捷伦6560液质联用设备中进行检测。本发明建立了一种新的液质联用检测方法,其能够检测出发酵蜂花粉中环氧肟酸、植物甾醇、氨基酸、阿魏酸衍生物这四种的具体含量,从而为以后发酵蜂花粉在预防前列腺增生上的研究提供了有效的支持。
Description
技术领域
本发明属于发酵蜂花粉成分检测领域,具体地说是一种发酵蜂花粉特殊成 分检测方法。
背景技术
随着社会的发展,人们的生活水平不断提高,对于食品的营养要求也不断 提高,本公司近年来主持研发《提高免疫力蜂花粉固态发酵产品的研究与开发》 的项目研发过程中发现发酵蜂花粉中含有环氧肟酸、植物甾醇、氨基酸、阿魏 酸衍生物这四种物质,其中脂质的为环氧肟酸、植物甾醇,水溶性的为氨基酸、 阿魏酸衍生物,这四种物质都具有预防前列腺增生的功效,但是由于目前没有 对发酵蜂花粉这四种物质的含量检测方法,故而无法得知各种发酵蜂花粉这四 种物质的含量多少,影响后续发酵蜂花粉在预防前列腺增生上的研究。
发明内容
本发明提供一种发酵蜂花粉特殊成分检测方法,用以解决现有技术中的缺 陷。
本发明通过以下技术方案予以实现:
一种发酵蜂花粉特殊成分检测方法,包括如下步骤:
步骤一:准确称取1g发酵蜂花粉,加100ml分析纯乙醇浸提1h,过滤,滤 渣再加入100ml分析纯乙醇浸提1h,过滤合并两次滤液并充分振荡均匀,用用 移液枪准确移取1ml的滤液加入50ml的容量瓶中,加色谱纯的甲醇定容,得待 检测液;
步骤二:将待检测液注入安捷伦6560液质联用设备中进行检测,其中,液 相色谱条件为:色谱柱为Agilent Eclipse Plus C18(2.1×100mm,1.8μm); 流动相:0.1%色谱纯甲酸水溶液,色谱纯甲醇;流速为0.3mL/min;梯度洗脱 条件:0min,95%0.1%色谱纯甲酸水溶液,5%色谱纯甲醇;1min,95%0.1% 色谱纯甲酸水溶液,5%色谱纯甲醇;10min,5%0.1%色谱纯甲酸水溶液,95% 色谱纯甲醇;13min,5%0.1%色谱纯甲酸水溶液,95%色谱纯甲醇;14min,100% 色谱纯甲醇;20min,100%色谱纯甲醇,21min,95%0.1%色谱纯甲酸水溶液, 5%色谱纯甲醇;质谱条件为:离子源:AIS ESI;检测模式为正离子模式;干燥 气流速为10L/min;干燥气温度320℃;雾化气流速为40psi;鞘气流速为 12L/min;鞘气温度为350℃;毛细管电压为3.5KV;喷嘴电压为0KV;扫描 范围:m/z 100-1200;参比离子:121.0509,922.0098。
如上所述的一种发酵蜂花粉特殊成分检测方法,所述的所的步骤二中0.1% 色谱纯甲酸水溶液中水为去离子水。
如上所述的一种发酵蜂花粉特殊成分检测方法,所述的步骤二中液相检测 时色谱柱柱温为40℃。
本发明的优点是:本发明建立了一种新的液质联用检测方法,其能够检测 出发酵蜂花粉中环氧肟酸、植物甾醇、氨基酸、阿魏酸衍生物这四种的具体含 量,从而为以后发酵蜂花粉在预防前列腺增生上的研究提供了有效的支持。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施 例或现有技术描述中所需要使用的附图作一简单地介绍,显而易见地,下面描 述中的附图是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出 创造性劳动性的前提下,还可以根据这些附图获得其他的附图。
图1是采用本发明方法检测发酵荷花粉的液质联用检测的液相图谱;
图2是采用本发明方法检测发酵荷花粉的液质联用检测的质谱图谱;
图3是采用本发明方法检测发酵油菜花粉的液质联用检测的液相图谱;
图4是采用本发明方法检测发酵油菜花粉的液质联用检测的质谱图谱;
图5是采用本发明方法检测发酵茶花粉的液质联用检测的液相图谱;
图6是采用本发明方法检测发酵茶花粉的液质联用检测的质谱图谱。
具体实施方式
为使本发明实施例的目的、技术方案和优点更加清楚,下面将结合本发明 实施例中的附图,对本发明实施例中的技术方案进行清楚、完整地描述,显然, 所描述的实施例是本发明一部分实施例,而不是全部的实施例。基于本发明中 的实施例,本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其 他实施例,都属于本发明保护的范围。
如图1,图3,图5所示,图中A区包含大量氨基酸,B区包含阿魏酸及其 衍生物,C包含赤霉素类化合物环氧肟酸,D包含植物甾醇;如图所示四者能够 很好的分离开来,同时通过对应的质谱图谱图2,图4,图6,根据图图1,图3, 图5的出峰顺序,判断对应峰是那种物质,从而得知对应物质的含量多少。
经过图谱对比水溶性物质氨基酸以及阿魏酸及其衍生物发现,三者后含有 大量的水溶性氨基酸,但仅在发酵油菜蜂花粉中富含阿魏酸及其衍生物,其他 两种发酵蜂花粉中阿魏酸衍生物含量较低,因此从水溶性成分组合来说建议只 选用发酵油菜蜂花粉作为产品开发对象。
经过图谱对比赤霉素类化合物环氧肟酸以及植物甾醇发现,发酵油菜蜂花 粉中富含大量的环氧肟酸和植物甾醇,其他两种发酵蜂花粉中这两类功能成分 含量较低,因此建议只选用发酵油菜蜂花粉作为产品开发对象。
通过上述检测分析可以得出,本发明的检测方法能快速且准确的得出被检 测发酵蜂花粉中环氧肟酸、植物甾醇、氨基酸、阿魏酸衍生物这四种物质的具 体含量,便于为后续研究开发打下坚实的基础。
最后应说明的是:以上实施例仅用以说明本发明的技术方案,而非对其限 制;尽管参照前述实施例对本发明进行了详细的说明,本领域的普通技术人员 应当理解:其依然可以对前述各实施例所记载的技术方案进行修改,或者对其 中部分技术特征进行等同替换;而这些修改或者替换,并不使相应技术方案的 本质脱离本发明各实施例技术方案的精神和范围。
Claims (3)
1.一种发酵蜂花粉特殊成分检测方法,其特征在于:包括如下步骤:
步骤一:准确称取1g发酵蜂花粉,加100ml分析纯乙醇浸提1h,过滤,滤渣再加入100ml分析纯乙醇浸提1h,过滤合并两次滤液并充分振荡均匀,用用移液枪准确移取1ml的滤液加入50ml的容量瓶中,加色谱纯的甲醇定容,得待检测液;
步骤二:将待检测液注入安捷伦6560液质联用设备中进行检测,其中,液相色谱条件为:色谱柱为Agilent Eclipse Plus C18(2.1×100mm,1.8μm);流动相:0.1%色谱纯甲酸水溶液,色谱纯甲醇;流速为0.3mL/min;梯度洗脱条件:0min,95%0.1%色谱纯甲酸水溶液,5%色谱纯甲醇;1min,95%0.1%色谱纯甲酸水溶液,5%色谱纯甲醇;10min,5%0.1%色谱纯甲酸水溶液,95%色谱纯甲醇;13min,5%0.1%色谱纯甲酸水溶液,95%色谱纯甲醇;14min,100%色谱纯甲醇;20min,100%色谱纯甲醇,21min,95%0.1%色谱纯甲酸水溶液,5%色谱纯甲醇;质谱条件为:离子源:AIS ESI;检测模式为正离子模式;干燥气流速为10L/min;干燥气温度320℃;雾化气流速为40psi;鞘气流速为12L/min;鞘气温度为350℃;毛细管电压为3.5KV;喷嘴电压为0KV;扫描范围:m/z 100-1200;参比离子:121.0509,922.0098。
2.根据权利要求1所述的一种发酵蜂花粉特殊成分检测方法,其特征在于:所述的所的步骤二中0.1%色谱纯甲酸水溶液中水为去离子水。
3.根据权利要求1所述的一种发酵蜂花粉特殊成分检测方法,其特征在于:所述的步骤二中液相检测时色谱柱柱温为40℃。
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Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4897224A (en) * | 1985-03-05 | 1990-01-30 | Morinaga Milk Industry Co., Ltd. | Method for producing ferulyl stanol derivatives |
US20040259732A1 (en) * | 2003-04-28 | 2004-12-23 | Monsanto Technology, L.L.C. | Treatment of plants and plant propagation materials with an antioxidant to improve plant health and/or yield |
CN108152402A (zh) * | 2017-12-18 | 2018-06-12 | 广西壮族自治区梧州食品药品检验所 | 液质联用检测中成药及保健食品中依达拉奉的方法 |
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Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4897224A (en) * | 1985-03-05 | 1990-01-30 | Morinaga Milk Industry Co., Ltd. | Method for producing ferulyl stanol derivatives |
US20040259732A1 (en) * | 2003-04-28 | 2004-12-23 | Monsanto Technology, L.L.C. | Treatment of plants and plant propagation materials with an antioxidant to improve plant health and/or yield |
CN108152402A (zh) * | 2017-12-18 | 2018-06-12 | 广西壮族自治区梧州食品药品检验所 | 液质联用检测中成药及保健食品中依达拉奉的方法 |
Non-Patent Citations (2)
Title |
---|
周艳明等: "油菜蜂花粉高效液相色谱指纹图谱的初步建立", 《食品安全与检测》 * |
李福高等: "花粉活性成分的提取与分离现状及展望", 《蜜蜂杂志》 * |
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