基质固相分散萃取-高效液相色谱法同时测定五味子中5种木脂素类化合物

doi:10.3724/SP.J.1123.2022.05012

摘要/Abstract

摘要：

研究建立了基质固相分散萃取-高效液相色谱(MSPD-HPLC)分析五味子中5种木脂素类化合物(五味子醇甲、五味子醇乙、五味子甲素、五味子乙素、五味子丙素)的方法。采用反相C₁₈色谱柱进行分离,以0.1%(v/v)甲酸水溶液和乙腈为流动相进行梯度洗脱,在波长250 nm下检测。考察了包括硅胶、酸性氧化铝、中性氧化铝、碱性氧化铝、佛罗里硅土、Diol、XAmide、Xion和C₁₈、C₁₈-ME、C₁₈-G₁、C₁₈-HC等在内的12种吸附剂以及吸附剂的质量、洗脱剂的种类、洗脱剂体积对五味子木脂素类化合物得率的影响。选定Xion作为MSPD-HPLC分析五味子中木脂素类化合物的吸附剂;基于吸附剂Xion的萃取参数优化结果表明:以0.25 g五味子粉末为固定值,Xion(0.75 g)为吸附剂,甲醇(15 mL)为洗脱剂,MSPD对五味子中木脂素类化合物具有较高的得率。建立的五味子中5种木脂素类化合物的分析方法,各目标分析物具有良好的线性关系(相关系数R²≥0.9999),检出限与定量限分别介于0.0089~0.0294 μg/mL和0.0267~0.0882 μg/mL之间。对五味子木脂素类化合物进行低、中、高3个水平的加标回收试验,平均回收率为92.2%~111.2%,相对标准偏差为0.23%~3.54%。日内和日间精密度均小于3.6%。与超声辅助提取和热回流提取前处理相比,MSPD具有萃取和净化相结合、耗时少、所需溶剂量少的优点,且MSPD-HPLC获得的结果优于经典方法。所建立的方法成功应用于17批五味子中5种木脂素类化合物含量的分析。

关键词: 基质固相分散萃取, 高效液相色谱, 木脂素类化合物, 五味子

Abstract:

The kidney-shaped, red-colord fruit from the plant, Schisandra chinensis (Turcz.) Baill, which belongs to the Schisandraceae family, is among the most popular remedies used in traditional Chinese medicine. The English name of the plant is “Chinese magnolia vine”. It has been used in Asia since ancient times to treat a variety of ailments, including chronic cough and dyspnea, frequent urination, diarrhea, and diabetes. This is because of the wide range of bioactive constituents, such as lignans, essential oils, triterpenoids, organic acids, polysaccharides, and sterols. In some cases, these constituents affects the pharmacological efficacy of the plant. Lignans with a dibenzocyclooctadiene-type skeleton are considered to be the major constituents and main bioactive ingredients of Schisandra chinensis. However, because of the complex composition of Schisandra chinensis, the extraction yields of lignans are low. Thus, it is particularly important to study pretreatment methods used during sample preparation for the quality control of traditional Chinese medicine. Matrix solid-phase dispersion extraction (MSPD) is a comprehensive process involving destruction, extraction, fractionation, and purification. The MSPD method is simple, it requires only a small number of samples and solvents, it does not require any special experimental equipments or instruments, and it can be used to prepare liquid, viscous, semi-solid, solid samples.

In this study, a method combining matrix solid-phase dispersion extraction with high performance liquid chromatography (MSPD-HPLC) was established for the simultaneous determination of five lignans (schisandrol A, schisandrol B, deoxyschizandrin, schizandrin B, and schizandrin C) in Schisandra chinensis. The target compounds were separated on a C₁₈ column with a gradient elution of 0.1% (v/v) formic acid aqueous solution and acetonitrile as the mobile phases, and detection was performed at a wavelength of 250 nm. First, the effects of 12 adsorbents, including silica gel, acidic alumina, neutral alumina, alkaline alumina, Florisil, Diol, XAmide, Xion, and the inverse adsorbents, C₁₈, C₁₈-ME, C₁₈-G₁, and C₁₈-HC, on the extraction yields of lignans were investigated. Second, effects of the mass of the adsorbent, the type of eluent, and volume of eluent on the extraction yields of lignans were investigated. Xion was chosen as an adsorbent for MSPD-HPLC analysis of lignans from Schisandra chinensis. Optimization of the extraction parameters showed that the MSPD method had a high lignan extraction yield with Schisandra chinensis powder (0.25 g) as a fixed value, Xion as the adsorbent (0.75 g), and methanol as the elution solvent (15 mL). Analytical methods were developed for five lignans from Schisandra chinensis and these methods showed good linearity (correlation coefficients (R²)≥ 0.9999) for each target analyte. The limits of detection and quantification ranged from 0.0089 to 0.0294 μg/mL and 0.0267 to 0.0882 μg/mL, respectively. Lignans were tested at low, medium, and high levels. The average recovery rates were 92.2% to 111.2%, and the relative standard deviations were 0.23% to 3.54%. Both intra-day and inter-day precisions were less than 3.6%. Compared with hot reflux extraction and ultrasonic extraction methods, MSPD has the advantages of combined extraction and purification, being less time-consuming, and requiring lower solvent volumes. Finally, the optimized method was successfully applied to analyze five lignans from Schisandra chinensis samples from 17 cultivation areas.

Key words: matrix solid-phase dispersion extraction (MSPD), high performance liquid chromatography (HPLC), lignans, Schisandra chinensis

中图分类号:

O658

杜昕昕, 王寅鹏, 肖伟, 朱靖博. 基质固相分散萃取-高效液相色谱法同时测定五味子中5种木脂素类化合物[J]. 色谱, 2023, 41(3): 257-264.

DU Xinxin, WANG Yinpeng, XIAO Wei, ZHU Jingbo. Simultaneous determination of five lignans from Schisandra chinensis by matrix solid-phase dispersion extraction-high performance liquid chromatography[J]. Chinese Journal of Chromatography, 2023, 41(3): 257-264.

图/表 10

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Compound	Linear range/(μg/mL)	Linear equation	R²	LOD/(μg/mL)	LOQ/(μg/mL)
Schisandrol A	0.429-268	Y=3.99×10⁷X+4.34×10⁴	0.9999	0.0098	0.0294
Schisandrol B	0.326-204	Y=3.20×10⁷X+1.94×10⁴	0.9999	0.0109	0.0327
Deoxyschizandrin	0.314-196	Y=3.92×10⁷X+2.34×10⁴	0.9999	0.0294	0.0882
Schizandrin B	0.320-200	Y=4.60×10⁷X+5.07×10⁴	0.9999	0.0089	0.0267
Schizandrin C	0.352-220	Y=2.91×10⁷X+1.78×10⁴	0.9999	0.0211	0.0633

Compound	Linear range/(μg/mL)	Linear equation	R²	LOD/(μg/mL)	LOQ/(μg/mL)
Schisandrol A	0.429-268	Y=3.99×10⁷X+4.34×10⁴	0.9999	0.0098	0.0294
Schisandrol B	0.326-204	Y=3.20×10⁷X+1.94×10⁴	0.9999	0.0109	0.0327
Deoxyschizandrin	0.314-196	Y=3.92×10⁷X+2.34×10⁴	0.9999	0.0294	0.0882
Schizandrin B	0.320-200	Y=4.60×10⁷X+5.07×10⁴	0.9999	0.0089	0.0267
Schizandrin C	0.352-220	Y=2.91×10⁷X+1.78×10⁴	0.9999	0.0211	0.0633

Compound	Background/(mg/mL)	Added/(mg/mL)	Found/(mg/mL)	Recovery/%	RSD/%
Schisandrol A	0.046	0.0300	0.074	94.7	0.32
		0.0600	0.105	97.5	1.34
		0.0900	0.146	111.2	0.71
Schisandrol B	0.013	0.0085	0.021	99.6	1.22
		0.0170	0.030	97.6	1.50
		0.0225	0.038	110.0	0.64
Deoxyschizandrin	0.010	0.0070	0.016	92.4	0.23
		0.0014	0.011	105.6	1.74
		0.0021	0.012	104.8	0.93
Schizandrin B	0.027	0.0125	0.039	94.5	0.34
		0.0250	0.050	92.2	2.12
		0.0375	0.068	109.1	0.81
Schizandrin C	0.0019	0.0015	0.003	95.0	0.78
		0.0030	0.005	99.3	1.60
		0.0045	0.006	98.4	3.54

Compound	Background/(mg/mL)	Added/(mg/mL)	Found/(mg/mL)	Recovery/%	RSD/%
Schisandrol A	0.046	0.0300	0.074	94.7	0.32
		0.0600	0.105	97.5	1.34
		0.0900	0.146	111.2	0.71
Schisandrol B	0.013	0.0085	0.021	99.6	1.22
		0.0170	0.030	97.6	1.50
		0.0225	0.038	110.0	0.64
Deoxyschizandrin	0.010	0.0070	0.016	92.4	0.23
		0.0014	0.011	105.6	1.74
		0.0021	0.012	104.8	0.93
Schizandrin B	0.027	0.0125	0.039	94.5	0.34
		0.0250	0.050	92.2	2.12
		0.0375	0.068	109.1	0.81
Schizandrin C	0.0019	0.0015	0.003	95.0	0.78
		0.0030	0.005	99.3	1.60
		0.0045	0.006	98.4	3.54

Compound	Extraction yields/(mg/g)
Compound	Hot reflux extraction	Ultrasonic extraction	MSPD
Schisandrol A	4.50	4.77	5.85
Schisandrol B	1.27	1.35	1.63
Deoxyschizandrin	1.02	1.11	1.34
Schizandrin B	3.11	3.15	3.70
Schizandrin C	0.21	0.24	0.32