赤芝中腺苷和尿苷HPLC指纹图谱及含量分析

doi:10.16178/j.issn.0528-9017.20240435

摘要/Abstract

摘要：

采用指纹图谱对赤芝水溶性成分中的腺苷和尿苷进行检测及含量测定,寻找不同来源孢子粉和子实体的含量变化规律,结果表明,不同来源和处理的灵芝粉和孢子粉在指纹图谱上有明显差异。赤芝菌盖中尿苷含量为0.072~0.849 mg·g^-1,菌柄中尿苷含量为0.056~0.302 mg·g^-1,破壁孢子粉中尿苷含量为0.032~0.089 mg·g^-1;赤芝菌盖中腺苷含量为0.019~0.118 mg·g^-1,菌柄中腺苷含量为0.005~0.024 mg·g^-1,破壁孢子粉中腺苷含量为0.024~0.107 mg·g^-1;同品种不同潮次采收的未破壁孢子粉主要成分含量无较大差异;同一品种的破壁孢子粉和未破壁孢子粉之间也无明显差异,但同一品种不同批次的赤芝中腺苷和尿苷含量差异显著。因此,可利用高效液相色谱法指纹图谱在8.5~20.0 min的区域内破壁孢子粉只有一个峰鉴别出破壁孢子粉,利用尿苷和腺苷的含量鉴别孢子粉中是否含有灵芝粉。

关键词: 赤芝, 高效液相色谱法指纹图谱, 水溶性物质, 腺苷, 尿苷

Abstract:

The fingerprint spectrum were used to detect and determine the content of adenosine and uridine in water-soluble components of Ganoderma lucidum,in order to find out the variation of the content of different spore powders and fruiting bodies. The results showed that there were significant differences in fingerprints between Ganoderma lucidum powder and spore powder from different sources; the content of uridine in the pileus of Ganoderma lucidum was 0.072-0.849 mg·g^-1, the content of uridine in the stipe was 0.056-0.302 mg·g^-1, and the content of uridine in the broken spore powder was 0.032-0.089 mg·g^-1; the content of adenosine in the pileus of Ganoderma lucidum was 0.019-0.118 mg·g^-1, the content of adenosine in the stipe was 0.005-0.024 mg·g^-1, and the content of adenosine in the broken spore powder is 0.024-0.107 mg·g^-1; There was no significant difference in the content of the main components of the unbroken spore powder harvested from the same variety but with different tides. There was no significant difference between the broken spore powder and the unbroken spore powder of the same variety. The difference of adenosine and uridine content in different batches of the same variety was obvious. The identification of broken spore powder can be achieved by using the fingerprint spectrum of high-performance liquid chromatography(HPLC)with only one peak in the area of 8.5-20.0 minutes for broken spore powder. The content of uridine and adenosine is used to identify whether the spore powder contains Ganoderma lucidum powder.

Key words: Ganoderma lucidum, high-performance liquid chromatography fingerprint spectrum, water-soluble substances, adenosine, uridine

中图分类号:

S567.3

林韧安, 李静静, 林琳, 黄爱云, 管楚韵, 章海英, 张伊涵, 林子琪, 刘京晶. 赤芝中腺苷和尿苷HPLC指纹图谱及含量分析[J]. 浙江农业科学, 2024, 65(8): 1764-1769.

LIN Ren'an, LI Jingjing, LIN Lin, HUANG Aiyun, GUAN Chuyun, ZHANG Haiying, ZHANG Yihan, LIN Ziqi, LIU Jingjing. HPLC fingerprint and content analysis of adenosine and uridine in Ganoderma lucidum[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(8): 1764-1769.

图/表 7

参考文献 23

[1]	国家药典委员会. 中华人民共和国药典-一部:2020年版[M]. 北京: 中国医药科技出版社, 2020.
[2]	武美华, 张胜男, 敬隆鑫, 等. 灵芝的活性成分及其药理作用的研究进展[J]. 中国林副特产, 2023(2):76-79.
[3]	何建明. 道藏集成-第一辑,1-正统道藏[M].[影印本]. 北京: 国家图书馆出版社, 2017.
[4]	QIN X J, FANG Z N, ZHANG J K, et al. Regulatory effect of Ganoderma lucidum and its active components on gut flora in diseases[J]. Frontiers in Microbiology, 2024, 15:1362479.
[5]	韩建军, 宁娜. 灵芝的化学成分与药理作用研究进展[J]. 广州化工, 2014, 42(23):18-19,29.
[6]	SEWERYN E, ZIAŁA A, GAMIAN A. Health-promoting of polysaccharides extracted from Ganoderma lucidum[J]. Nutrients, 2021, 13(8): 2725.
[7]	MISHRA J, JOSHI A, RAJPUT R, et al. Phenolic rich fractions from Mycelium and fruiting body of Ganoderma lucidum inhibit bacterial pathogens mediated by generation of reactive oxygen species and protein leakage and modulate hypoxic stress in HEK 293 cell line[J]. Advances in Pharmacological Sciences, 2018, 2018: 6285615.
[8]	郑洁, 唐庆九, 张劲松, 等. 灵芝子实体中三萜类物质抗肿瘤活性的谱效关系[J]. 菌物学报, 2019, 38(7):1165-1172.
[9]	高德勐, 付钰涓, 蒲馨怡, 等. 基于网络药理学和分子对接技术的灵芝改善2型糖尿病作用机制[J]. 北华大学学报(自然科学版), 2024, 25(3):339-345.
[10]	CHEN Y S, CHEN Q Z, WANG Z J, et al. Anti-inflammatory and hepatoprotective effects of Ganoderma lucidum polysaccharides against carbon tetrachloride-induced liver injury in Kunming mice[J]. Pharmacology, 2019, 103(3/4):143-150.
[11]	AHMAD M F, AHMAD F A, ZEYAULLAH M, et al. Ganoderma lucidum: novel insight into hepatoprotective potential with mechanisms of action[J]. Nutrients, 2023, 15(8):1874.
[12]	LIU X J, YANG L D, LI G Y, et al. A novel promising neuroprotective agent: Ganoderma lucidum polysaccharide[J]. International Journal of Biological Macromolecules, 2023, 229: 168-180.
[13]	丰彬彬, 孔佳欣, 卫彩红, 等. 灵芝的药用成分及相关应用研究[J]. 中国食品, 2023(22):85-87.
[14]	赵小平. 药用真菌灵芝研究与栽培现状[J]. 中国食用菌, 2019, 38(7):1-5.
[15]	陈体强, 李开本, 陈朝旭, 等. 灵芝、紫芝担孢子及其孢壁的超微结构[J]. 福建农业学报, 1998, 13(4):34-39.
[16]	李玲, 孙元章, 李刚. 灵芝生物活性成分及其药理作用研究进展[J]. 南方农业, 2019, 13(4):50-55.
[17]	陈军辉, 吴凤琪, 李文龙, 等. 灵芝孢子粉与子实体中三萜类化合物、多糖及重金属元素的含量比较[J]. 中国药房, 2008, 19(33):2585-2587.
[18]	张忠, 张劲松, 刘艳芳, 等. 分光光度法测定灵芝中总三萜含量方法探讨[J]. 上海农业学报, 2016, 32(1):61-65.
[19]	王健, 焦强, 王海波, 等. 灵芝孢子粉质量分析方法研究进展[J]. 药物分析杂志, 2016, 36(5):749-755.
[20]	王美丹, 胡扬, 宋辉, 等. 灵芝孢子粉化学成分和药理作用的研究进展及其质量标志物(Q-Marker)预测分析[J]. 中草药, 2023, 54(23):7918-7933.
[21]	杨吉, 冼玲, 钟天圣, 等. 近红外光谱指纹图谱的灵芝品种快速鉴别方法[J]. 时珍国医国药, 2017, 28(6):1359-1361.
[22]	李静静, 刘京晶, 斯金平, 等. 赤芝不同部位油脂组成分析[J]. 中国中药杂志, 2015, 40(14):2814-2819.
[23]	吕明亮, 斯金平, 应国华, 等. 一种灵芝孢子粉收集器的研究[J]. 中国中药杂志, 2009, 34(5): 539-541.

标准品	标准曲线方程	r	线性范围/μg
尿苷	y=48.515x+5.886 7	1.000	0.20~2.00
腺苷	y=2.113 4x+0.121 8	1.000	0.02~0.20

标准品	标准曲线方程	r	线性范围/μg
尿苷	y=48.515x+5.886 7	1.000	0.20~2.00
腺苷	y=2.113 4x+0.121 8	1.000	0.02~0.20

标准品	样品中量/μg	加入量/ μg	测得量/ μg	回收率/ %	平均回收率/%	RSD/ %
尿苷	11.68	9.20	20.92	100.43	99.89	0.79
	11.63	9.20	20.75	99.13
	11.66	9.20	20.83	99.67
	11.59	11.60	23.36	101.47
	11.65	11.60	23.11	98.79
	11.64	11.60	23.17	99.40
	11.73	14.00	25.71	99.86
	11.72	14.00	25.75	100.21
	11.68	14.00	25.69	100.07
腺苷	8.93	7.20	16.17	100.53	99.30	1.11
	8.94	7.20	16.06	98.85
	8.92	7.20	16.11	99.79
	8.94	9.00	17.82	98.58
	8.93	9.00	17.69	97.26
	8.93	9.00	17.93	99.94
	8.95	10.80	19.58	98.39
	8.98	10.80	19.77	99.87
	8.93	10.80	19.79	100.50

标准品	样品中量/μg	加入量/ μg	测得量/ μg	回收率/ %	平均回收率/%	RSD/ %
尿苷	11.68	9.20	20.92	100.43	99.89	0.79
	11.63	9.20	20.75	99.13
	11.66	9.20	20.83	99.67
	11.59	11.60	23.36	101.47
	11.65	11.60	23.11	98.79
	11.64	11.60	23.17	99.40
	11.73	14.00	25.71	99.86
	11.72	14.00	25.75	100.21
	11.68	14.00	25.69	100.07
腺苷	8.93	7.20	16.17	100.53	99.30	1.11
	8.94	7.20	16.06	98.85
	8.92	7.20	16.11	99.79
	8.94	9.00	17.82	98.58
	8.93	9.00	17.69	97.26
	8.93	9.00	17.93	99.94
	8.95	10.80	19.58	98.39
	8.98	10.80	19.77	99.87
	8.93	10.80	19.79	100.50

部位	龙芝1号 1潮	龙芝1号 2潮	沪农1号 1潮	沪农1号 2潮
菌盖	0.894	0.996	0.956	0.495
菌柄	0.866	0.994	0.886	0.595
破壁孢子粉	0.968	0.988	0.946	0.894
未破壁孢子粉	0.982	0.991	0.995	0.995