食品科学 ›› 2023, Vol. 44 ›› Issue (2): 312-318.doi: 10.7506/spkx1002-6630-20211203-039

• 成分分析 • 上一篇    

诺邓火腿红色素的分离纯化及结构鉴定

杨子江,张丽红,廖国周,田梅,吕东霖,何颖,葛长荣,王桂瑛   

  1. (1.云南农业大学食品科学技术学院,云南 昆明 650201;2.云南农业大学 云南省畜产品加工工程技术研究中心,云南 昆明 650201)
  • 发布日期:2023-01-31
  • 基金资助:
    云南省中青年学术和技术带头人后备人才项目(202105AC160068);云南省科技计划项目重大科技专项(2019ZG003)

Isolation, Purification and Structural Identification of Red Pigment from Nuodeng Ham

YANG Zijiang, ZHANG Lihong, LIAO Guozhou, TIAN Mei, LÜ Donglin, HE Ying, GE Changrong, WANG Guiying   

  1. (1. College of Food Science and Technology, Yunnan Agricultural University, Kunming 650201, China; 2. Livestock Product Processing and Engineering Technology Research Center of Yunnan Province, Yunnan Agricultural University, Kunming 650201, China)
  • Published:2023-01-31

摘要: 为探究诺邓火腿红色素化学本质,采用75%丙酮溶液提取,结合C18固相萃取小柱进行分离纯化,通过紫外-可见光光谱(ultraviolet spectroscopy,UV-Vis)、荧光光谱、超高效液相色谱-串联质谱(ultra-high performance liquid chromatography-tandem mass spectrometry,UPLC-MS/MS)、傅里叶变换红外光谱(Fourier transform infrared,FTIR)和核磁共振(nuclear magnetic resonance,NMR)共同表征诺邓火腿红色素的化学结构。结果表明:在UV-Vis中416 nm处有1 个强吸收峰,546 nm和584 nm处有2 个弱对称吸收峰,符合金属卟啉特征;以420 nm激发,红色素在590 nm处有1 个强荧光发射峰,644 nm处有1 个弱荧光发射峰,与Zn-原卟啉IX(Zn protoporphyrin IX,ZnPP)标准品比对后高度重合,表明卟啉环中的金属离子是锌离子;在UPLC-MS/MS的正离子(m/z 625.177 9 [M+H]+)和负离子(m/z 623.161 4 [M-H]-)模式下均检测出ZnPP的存在;在FTIR中发现—CH3、—CH2—、C=O、羧基中—C—OH、烯烃上的C-H和卟啉环C=N等卟啉环上的特征官能团伸缩振动;分析诺邓火腿红色素的1H-NMR,发现与其目标化合物ZnPP结构一致,因此鉴定出诺邓火腿红色素的化学本质为ZnPP。研究结果可以为诺邓火腿色泽调控提供一定理论依据。

关键词: 诺邓火腿;红色素;分离纯化;Zn-原卟啉IX;结构表征

Abstract: In order to explore the chemical nature of red pigment in Nuodeng ham, the red pigment was extracted with 75% acetone aqueous solution and purified using a C18 solid phase extraction column, and its chemical structure was characterized by using ultraviolet-visible (UV-Vis) spectroscopy, fluorescence spectroscopy and ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS), Fourier transform infrared (FTIR) spectroscopy and nuclear magnetic resonance (NMR) spectroscopy. The UV-Vis spectrum showed a strong absorption peak at 416 nm and two weakly symmetrical absorption peaks at 546 and 584 nm, which are characteristic of metalloporphyrins. When it was excited at 420 nm, the red pigment exhibited a strong fluorescence emission peak at 590 nm and a weak fluorescence emission peak at 644 nm, which highly coincided with those of Zn protoporphyrin IX (ZnPP) standard, indicating that the metal ions in the porphyrin ring were zinc ions. ZnPP was detected by UPLC-MS/MS in both positive (m/z 625.177 9 [M + H]+) and negative ion (m/z 623.161 4 [M-H]–) modes, and the stretching vibrations of characteristic functional groups on the porphyrin ring such as –CH3, –CH2–, C=O, –C–OH in carboxyl groups, C–H on olefin and porphyrin ring C=N were observed in the FTIR spectrum. In addition, the 1H-NMR results of the red pigment were consistent with the structure of the target compound ZnPP. Therefore, the red pigment in Nuodeng ham was identified as ZnPP. The results of this study provide a theoretical basis for the color regulation of Nuodeng ham.

Key words: Nuodeng ham; red pigment; separation and purification; Zn protoporphyrin IX; structural characterization

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