4种大型红藻类菌胞素氨基酸的提取工艺优化与分离鉴定

类菌胞素氨基酸（Mycosporine-like amino acids，MAAs）广泛存在于大型红藻中，是一类具有良好应用前景的活性物质。本文采用单因素和正交试验，分析了提取温度、时间、次数和料液比对红毛苔、石花菜、菊花江蓠和江蓠中MAAs提取物得率的影响，优化提取MAAs工艺，并对提取物进行了分离鉴定。结果表明，毛苔、石花菜、菊花江蓠和江蓠中MAAs优化提取工艺分别为:45 ℃、3 h、4次、1:25 g/mL；45 ℃、1 h、4次、1:20 g/mL；45 ℃、2 h、3次、1:15 g/mL；40 ℃、1 h、3次、1:20 g/mL。应用上述提取工艺，制备得到4种大型红藻MAAs提取物，提取物得率依次为249.3、197.9、146.4和449.5 mg/g，其在紫外光谱和薄层层析板呈现MAAs紫外吸收特性和颜色反应。同时，采用硅胶柱层析，对红毛苔和江蓠MAAs提取物进行分离，共得到3个组分H1、J1和J2。通过紫外光谱扫描、高效液相色谱和质谱分析，并与已有文献比较，确定了其MAAs组成。H1包括shinorine、palythine和porphyra-334（3种MAA含量为组分H1含量的95.4%）、palythenic acid（4.6%）；J1包括shinorine、palythine和porphyra-334（3种MAA 为组分J1含量的96.3%）、palythenic acid（3.7%）；J2为palythine单体。

Optimization of Extraction Process,Isolation and Characterization of Mycosporine-Like Amino Acids from Four Species of Marine Red Macroalgaes

Marine macroalgae is one of the good sources of mycosporine-like amino acids (MAAs), especially red macroalgae. As a new type of active substances with commercial development prospect, extraction, isolation and characterization of MAAs are far from enough for the application. To obtain the extraction process of MAAs from four species of red macroalgae (Bangia fusco-purpurea, Gelidium amansii, Gracilaria confervoides, and Gracilaria sp.), a series of single-factor and orthogonal experiments were carried out to analyze the effects of extraction temperature, time, times and solid-liquid ratio on the yields of MAAs. The extraction process of MAAs was optimized, and the extracts were isolated and identified. The optimized extraction processes of MAAs were obtained as follows: 45 ℃, 3 h, 4 times, 1:25 g/mL. 45 ℃, 1 h, 4 times, 1:20 g/mL. 45 ℃, 2 h, 3 times, 1:15 g/mL. 40 ℃, 1 h, 3 times, 1:20 g/mL. Four MAAs extracts of red algae were prepared by using the above extraction process, and the yields were 249.3, 197.9, 146.4 and 449.5 mg/g, respectively. All of them showed UV absorption characteristics and color reaction of MAAs in UV spectrum and thin layer chromatography (TLC). Furthermore, MAAs extracts from Bangia fusco-purpurea (or Gracilaria sp.) were isolated by silica gel column chromatography to obtain one fraction H1 (or two fractions J1 and J2). MAAs composition and proportion of these fractions were determined by UV, HPLC and MS, and comparison with existing literatures. H1 included shinorine, palythine and porphyra-334 (the content of these three MAAs was 95.4% of the content of the fraction H1), and palythenic acid (4.6%). Shinorine, palythine, porphyra-334 (96.3% of the fraction J1), and palythenic acid (3.7%) were in J1. And J2 was palythine.