目前对佛手(Citrus medica L.var.sarcodactylis Swingle)次生代谢产物筛查鉴定的物质种类偏少且缺乏系统的定量分析。该文采用超高效液相色谱串联三重四级杆质谱法(ultra-high performance liquid chromatography coupled with triple-quadrupole mass spectrometry, UPLC-QqQ-MS/MS)同时对金华和云南两地的佛手中36种类黄酮和23种香豆素类物质进行靶向筛查,共检出了36种类黄酮和21种香豆素类物质,并对其中的34种类黄酮和21种香豆素进行了准确定量,有2种物质能检出但不能准确定量,这是目前对佛手所含次生代谢产物进行定量分析物质数量最多的研究。结果显示,金华佛手和云南佛手中所测类黄酮总含量分别为(757.11±27.12) μg/g和(803.98±29.41) μg/g,香豆素类物质总含量分别为(1 102.72±13.29) μg/g和(676.97±49.77) μg/g。地奥司明是金华佛手和云南佛手含量最高的类黄酮,分别高达(345.85±19.92) μg/g和(417.73±12.13) μg/g,其次是橙皮苷,含量分别为(201.93±1.07) μg/g和(271.78±13.08) μg/g。川陈皮素和枸橘苷在2种佛手中均有检出,但不能准确定量,栀子黄素B在金华佛手中有检出,但不能准确定量,在云南佛手中的含量也低于0.1 μg/g;柠檬内酯(5,7-二甲氧基香豆素)是2种佛手中含量最高的香豆素类物质,分别高达(946.17±10.78) μg/g和(561±46.42) μg/g,花椒毒素和异补骨脂素在2种佛手中均无检出。此外,有14种类黄酮和4种香豆素首次在佛手中检出,包括牡荆素鼠李糖苷、5-羟基-7,8,4′-三甲氧基黄酮、半齿泽兰素-5-甲醚、根皮素、根皮苷、蔓荆子黄素、异橙黄酮、4′,5,7,8-四甲氧基黄酮、4′,5,6,7-四甲氧基黄酮、芹菜素、栀子黄素B、栎精-3,7,3′,4′-四甲醚、金合欢素、3′,4′,5,5′,6,7-六甲氧基黄酮以及6′,7′-环氧香柠檬亭、6′,7′-二羟基香柠檬亭、5-牻牛儿醇基-7-甲氧基香豆素和异紫花前胡内酯。研究结果将为佛手类黄酮和香豆素类次生代谢的相关数据库提供数据支撑,为佛手种质资源的开发利用提供参考。
The flavonoids and coumarins in Jinhua fingered citron and Yunnan fingered citron (Citrus medica L. var. sarcodactylis Swingle) were screened and quantitated based on the ultra-high performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UPLC-QqQ-MS/MS) method contained 36 flavonoids and 23 coumarins. A total of 34 kinds of flavonoids and 21 kinds of coumarins were detected and accurately quantitated. This is a study with the maximum number of compounds among the quantitative analysis of secondary metabolites quantitated for fingered citron at present. The total flavonoid contents of Jinhua and Yunnan fingered citrons were (757.11±27.12) μg/g and (803.98±29.41) μg/g, respectively. The contents of total coumarins were (1 102.72±13.29) μg/g and (676.97±49.77) μg/g, respectively. Diosimin was the flavonoid with the highest content of (345.85±19.92) μg/g and (417.73±12.13) μg/g, respectively in Jinhua and Yunnan fingered citrons, followed by hesperidin, with contents of (201.93±1.07) μg/g, (271.78±13.08) μg/g, respectively. Nobiletin and poncirin were detected in two kinds of fingered citrons, but they could not be quantitated accurately. In Jinhua fingered citron, gardenin B was detected but could not be accurately quantitated. And in Yunnan fingered citron, its content was also lower than 0.1 μg/g. Limettin was the coumarin with the highest content in the two fingered citrons as high as (946.17±10.78) μg/g and (561±46.42) μg/g, respectively. Xanthotoxin and isopsoralen were not detected in the two fingered citrons. In addition, 14 kinds of flavonoids including vitexin-2-O-rhamnoside, 5-hydroxy-7,8,4′-trimethoxylflavone, eupatorin-5-methylether, phloretin, phlorizin, vitexicarpin, isosinensetin, 4′,5,7,8-tetramethoxylflavone, 4′,5,6,7-tetramethoxylflavone, apigenin, gardenin B, 5-hydroxy-3,7,3′,4′-tetramethoxyflavone, acacetin, and 3′,4′,5,5′,6,7-hexathoxylflavone, and 4 coumarins including 6′,7′-epoxybergamottin, 6′,7′-dihydroxybergamottin, 5-geranyloxy-7-methoxycoumarin, and marmesin were first detected in fingered citrons. Results could provide data support for the related database of flavonoid and coumarin metabolism for fingered citrons and provide a reference for the development and utilization of germplasm resources of fingered citrons.
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