|
|
高效液相色谱检测红球藻中虾青素的技术研究
|
Abstract:
优化高效液相色谱法检测红球藻中虾青素的条件和方法。使用乙醇–甲醇(1:3)提取红球藻样品，经0.1 mol/L氢氧化钠–甲醇皂化后，用0.2 mL 2%磷酸–甲醇中和剩余碱后，采用高效液相色谱法测定。总虾青素在0.1~10 μg/mL范围内线性良好(R2 ≥ 0.999)，方法定量限10 mg/kg。在添加水平为10 mg/kg和50 mg/kg时，回收率为91%~100%，相对标准偏差低于1.0%。该方法较GB/T 31520-2015中使用甲醇–叔丁基甲醚–磷酸为流动相，二氯甲烷–甲醇(1:3)作为提取液，经0.1 mol/L氢氧化钠–甲醇皂化后，用0.4 mL 2%磷酸–甲醇中和剩余碱后，结果更准确、稳定、灵敏，且更为环保，能够满足红球藻中总虾青素检测与确证需要。
To optimize the conditions and methods for the determination of astaxanthin in Haematococcus by high-performance liquid chromatography (HPLC). The Haematococcus samples were extracted using ethanol-methanol (1:3), saponified by 0.1 mol/L sodium hydroxide-methanol, and then determined by high-performance liquid chromatography (HPLC) after neutralization of the remaining alkali with 0.2 mL 2% phosphoric acid-methanol. The linearity of total astaxanthin was good in the range of 0.1~10 μg/mL (R2 ≥ 0.999), and the limit of quantification of the method was 10 mg/kg. The recoveries ranged from 91% to 100% at the spiked levels of 10 mg/kg and 50 mg/kg with the relative standard deviations (RSDs) lower than 1.0%. Compared with GB/T 31520-2015, the method used methanol-tert-butyl-methyl ether-phosphoric acid as mobile phase, dichloromethane-methanol (1:3) as extraction liquid, was saponified by 0.1mol/L sodium hydroxide-methanol, and neutralized by 0.4mL 2% phosphoric acid-methanol, the results were more accurate, stable, sensitive, and environmentally friendly. It can meet the need for the detection and confirmation of total astaxanthin in Haematococcus.
| [1] | 赵英源, 刘俊霞, 陈姝彤, 等. 虾青素生理活性的研究进展[J]. 中国海洋药物, 2020, 39(3): 80-88. |
| [2] | 江利华, 柳慧芳, 郝光飞, 等. 虾青素抗氧化能力研究进展[J]. 食品工业科技, 2019, 40(10): 350-354. |
| [3] | 孟昂, 赵晓燕, 朱运平, 等. 天然色素虾青素的功能性研究进展[J]. 粮油食品科技, 2019, 27(5): 49-54. |
| [4] | Chintong, S., Phatvej, W., Rerk-Am, U., et al. (2019) In vitro Antioxidant, Antityrosinase, and Cytotoxicactivities of Astaxanthin from Shrimp Waste. Antioxidants, 8, Article 128. https://doi.org/10.3390/antiox8050128 |
| [5] | 李勇超, 贺青华, 刘瑞雪, 等. 雨生红球藻源虾青素对糖尿病小鼠的降糖作用及其机制[J]. 食品工业科技, 2016, 37(24): 355-359. |
| [6] | Wang, J.H., Liu, S.W., Wang, H., et al. (2019) Xanthophyllomyces dendrorhous-Derived Astaxanthin Regulates Lipid Metabolism and Gut Microbiota in Obese Mice Induced by a High-Fat Diet. Marine Drugs, 17, Article 337. https://doi.org/10.3390/md17060337 |
| [7] | 左正宇, 邵阳, 刘杨, 等. 虾青素调节肝脂代谢与昼夜节律基因表达[J]. 食品科学, 2019, 40(3): 165-172. |
| [8] | Lin, W.N., Wen, Y.T. and Tsai, R.K. (2018) Neuroprotective Effect of Astaxanthin in a Rat Model of Anterior Ischemic Optic Neuropathy. Investigative Ophthalmology & Visual Science, 59, Article 2498. |
| [9] | Dong, S.Z., Huang, Y., Zhang, R., et al. (2014) Four Different Methods Comparison for Extraction of Astaxanthin from Green Alga Haematococcus pluvialis. The Scientific World Journal, 2014, Article ID: 694305. https://doi.org/10.1155/2014/694305 |
| [10] | 班磊, 尤建伟. 虾青素制备方法的研究进展[J]. 化工管理, 2019(21): 31-32. |
| [11] | 陈丹, 汪锋, 蒋珊, 等. 虾青素化学和生物合成研究进展[J]. 食品工业科技, 2021, 42(21): 445-453. |
| [12] | 于广利, 赵峡编. 糖药物学[M]. 青岛: 中国海洋大学出版社, 2012. |
| [13] | 陈涛, 邵泽彻, 杨福梅, 等. 雨生红球藻中天然虾青素的应用研究[J]. 农业工程技术, 2023, 43(15): 76-77. |
| [14] | 余小元, 谢子健, 曹月琴, 等. 虾青素的主要来源物种——雨生红球藻[J]. 生物学通报, 2022, 57(2): 7-11. |
| [15] | 黄静雯, 谭惠文, 林怡妤, 等. 虾青素毒性的研究图谱分析及安全性评价[J]. 广东化工, 2023, 50(8): 102-104. |
| [16] | 李艳, 高静. 雨生红球藻天然虾青素提取研究进展[J]. 食品与机械, 2020, 36(12): 211-217. |
| [17] | 中华人民共和国国家卫生和计划生育委员会. GB 2760-2014食品安全国家标准食品添加剂使用标准[S]. 北京: 标准出版社, 2014. |
| [18] | 中华人民共和国国家质量监督检验检疫总局, 中国国家标准化管理委员会. GB/T 31520-2015红球藻中虾青素的测定液相色谱法[S]. 北京: 标准出版社, 2015. |
| [19] | 中华人民共和国国家质量监督检验检疫总局, SN/T 2327-2009进出口动物源性食品中角黄素、虾青素的检测方法[S]. 北京: 标准出版社, 2009. |
| [20] | 张泳, 厉妙沙, 吴嘉圣. 分光光度计法测定虾青素[J]. 浙江化工, 2015, 46(9): 51-54. |
| [21] | 赵立艳, 赵广华, 陈芳, 等. 利用高效液相色谱法测定雨生红球藻中虾青素含量[J]. 食品工业科技, 2006, 27(5): 177-178. |
| [22] | 雷凤爱. 雨生红球藻中虾青素的分离纯化及异构体的分析研究[D]: [硕士学位论文]. 呼和浩特: 内蒙古工业大学, 2009. |
