超声辅助结合离子液体双水相提取-高效液相色谱法测定三七中5种稀有人参皂苷的含量

doi:10.7503/cjcu20150703

摘要/Abstract

摘要：

建立了超声辅助结合离子液体双水相提取-高效液相色谱法测定三七中5种稀有人参皂苷20(S)-Rg₂, 20(S)-Rh₁, Rk₃, 20(S)-Rg₃及Rk₁含量的方法. 考察了影响提取率的多种因素并确立了最佳提取条件, 在该条件下, 5种稀有人参皂苷加样回收率均在92.07%~110.55%之间, RSD值均<5.43%. 实验结果表明, 该方法具有快速、高效、准确且环保等优点, 为中药材及其制剂中微量化学成分的提取、富集与分析提供了参考.

关键词: 离子液体, 超声辅助提取, 三七, 稀有人参皂苷

Abstract:

A method of ultrasonic-assisted ionic liquid-salt aqueous two-phase extraction coupled with high-performance liquid chromatography(HPLC) was developed for the determination of the contents of five rare ginsenosides[20(S)-Rg₂, 20(S)-Rh₁, Rk₃, 20(S)-Rg₃ and Rk₁] in Panax notoginseng(Burk.) F.H.Chen(Sanqi). The factors influencing the ultrasonic-assisted extraction were investigated in detail. Under the optimized conditions, satisfactory extraction efficiency was achieved with the recoveries ranging from 92.07% to 110.55%, and the RSDs were lower than 5.43%. The results showed that the method had the advantages of fast, efficient, accurate and environmental protection. It provided a strong basis for the extraction, enrichment and analysis of the trace bio-active chemical composition of the traditional Chinese medicine and its preparations.

Key words: Ionic liquid, Ultrasonic-assisted extraction, Panax notoginseng(Burk.) F.H.Chen, Rare ginsenoside

中图分类号:

O657.7

TrendMD:

李兰杰, 李绪文, 丁健, 刘迎, 吴谦, 王晓中, 李敏, 金永日. 超声辅助结合离子液体双水相提取-高效液相色谱法测定三七中5种稀有人参皂苷的含量. 高等学校化学学报, 2016, 37(3): 454.

LI Lanjie, LI Xuwen, DING Jian, LIU Ying, WU Qian, WANG Xiaozhong, LI Min, JIN Yongri. Ionic Liquid Surfactant-mediated Ultrasonic-assisted Extraction Coupled with HPLC for the Determination of Five Rare Ginsenosides in Panax notoginseng (Burk.) F. H. Chen^†. Chem. J. Chinese Universities, 2016, 37(3): 454.

图/表 9

Fig.1 Chemical structures of rare ginsenosides 20(S)-Rg2, 20(S)-Rh1, 20(S)-Rg3, Rk1 and Rk3

Fig.2 Chromatograms of standards(a) and rhizome of Sanqi(GJ-1)(b)

Fig.3 Effects of different degreasants on the extraction efficiencyConditions: 1.00 mL ionic liquid, 5.0 mL water, 7.00 g K2HPO4, 0.60 g Sanqi, ultrasonic time was 20 min. Error bars represent SDs.

Fig.4 Effects of different amounts of K2HPO4 on the extraction efficiency Conditions: 1.00 mL ionic liquid, 5.0 mL water.

Fig.5 Effects of different amounts of Sanqi on the extraction efficiencies of 20(S)-Rg2(a) and 20(S)-Rh1(b)The degreasant was petroleum ether. Conditions: 1.00 mL ionic liquid, 5.0 mL water, 7.00 g K2HPO4, ultrasonic time was 20 min. Each data point represents two analytes.

Fig.6 Effects of the ultrasonic treatment time on the extraction efficiencies of 20(S)-Rg2(a) and 20(S)-Rh1(b)The degreasant was petroleum ether. Conditions: 1.00 mL ionic liquid. 5.0 mL water, 7.00 g K2HPO4, 0.60 g Sanqi. Each data point represents two analytes. Error bars represent SDs.

Table 1 Regression equations, LODs, LOQs, intra-day and inter-day precision

Analyte	Linear regression	Linear range/ (μg·mL^-1)	r²	Intra-day precision(%)	Inter-day precision(%)	LOD/ (μg·mL^-1)	LOQ/ (μg·mL^-1)
20(S)-Rg₂	y=9.30x+9.55	4.05—1012	0.9998	0.98	1.03	0.32	1.28
20(S)-Rh₁	y=11.17x+8.52	5.28—528.0	0.9996	1.13	1.07	0.21	0.80
Rk₃	y=25.30x+10.78	5.09—508.8	0.9998	1.65	1.63	0.18	7.02
20(S)-Rg₃	y=9.30x+9.82	5.04—504.0	0.9998	1.23	1.33	0.48	1.98
Rk₁	y=10.80x+3.61	3.54—353.6	0.9999	0.89	1.03	0.29	0.97

Table 2 Recoveries of the analytes(N=6)

Analyte	Initial content/(μg·mL^-1)	Added/(μg·mL^-1)	Found/(μg·mL^-1)	Recovery(%)	RSD(%, N=6)
20(S)-Rg₂	506.20±0.70	506.00	1006.30	99.42	3.08
20(S)-Rh₁	90.47±0.58	99.24	180.67	95.23	3.87
Rk₃	5.51±0.09	7.95	14.88	110.55	5.43
20(S)-Rg₃	95.93±0.32	63.00	160.36	100.90	2.38
Rk₁	10.65±0.01	11.05	19.98	92.07	4.43

Table 3 Contents(%) of the rare ginsenosides in Sanqi(N=6)

Sample	Content(%)
Sample	20(S)-Rg₂	20(S)-Rh₁	Rk₃	20(S)-Rg₃	Rk₁
GJ-1	0.1181±0.0033	0.02111±0.00045	0.001287±0.000035	0.02238±0.00024	0.002485±0.000025
GJ-2	0.1178±0.0034	0.02056±0.00053	0.001406±0.000039	0.02292±0.00022	0.002540±0.000032
JT-1	0.1120±0.0043	0.01768±0.00039	0.001271±0.000042	0.02238±0.00024	0.002730±0.000031

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