A simple and sensitive HPLC method for the simultaneous determination of eight bioactive components and fingerprint analysis of Schisandra sphenanthera

Department of Pharmacy, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China.
Analytica chimica acta (Impact Factor: 4.51). 03/2010; 662(1):97-104. DOI: 10.1016/j.aca.2009.12.039
Source: PubMed


A simple and sensitive high performance liquid chromatography method with photodiode array detection (HPLC-DAD) was developed for simultaneous determination of eight bioactive constituents (schisandrin, schisandrol B, schisantherin A, schisanhenol, anwulignan, deoxyshisandrin, schisandrin B and schisandrin C) in the ripe fruit of Schisandra sphenanthera and its traditional Chinese herbal preparations Wuzhi-capsule by optimizing the extraction, separation and analytical conditions of HPLC-DAD. The chemical fingerprint of S. sphenanthera was established using raw materials of 15 different origins in China. The chromatographic separations were obtained by an Agilent Eclipse XDB-C18 reserved-phase column (250 mm x 4.6 mm i.d., 5 microm) using gradient elution with water-formic acid (100:0.1, v/v) and acetonitrile, at a flow rate of 1.0 mL min(-1), an operating temperature of 35 degrees C, and a wavelength of 230 nm. The constituents were confirmed by (+) electrospray ionization LC-MS. The new method was validated and was successfully applied to simultaneous determination of components in 13 batches of Wuzhi-capsule. The results indicate that this multi-component determination method in combination with chromatographic fingerprint analysis is suitable for quantitative analysis and quality control of S. sphenanthera.

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Available from: Wan-Sheng Chen, Nov 06, 2014
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    • "In the process, techniques such as HPLC, (gas chromatography) GC, gas chromatography-mass spectrometry (GC-MS), and liquid chromatography-mass spectrometry (LC-MS) are often used. However, HPLC is simple, reliable, and inexpensive, and has been widely used for quantitative analysis of herbal medicine.[1112131415] "
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    ABSTRACT: Background:Tilia amurensis consists of various compounds, such as flavonoids and terpenoids.Objective:A simple and reliable high performance liquid chromatography (HPLC) coupled with the diode array detector (DAD) method has been established for simultaneous determination of epicatechin, nudiposide, lyoniside, and scopoletin isolated from Tilia amurensis.Materials and Methods:Optimum separations were obtained with a SHISEIDO C18 column by gradient eluton, with 0.1% Trifluoroacetic acid (TFA) water-methanol as the mobile phase. The gradient elution system was completed within 40 minutes. The flow rate and detection wavelength were 1 mL/minute, 205 nm, 250 nm, and 280 nm, respectively.Results:Validation of the analytical method was evaluated by linearity, precision, and the accuracy test. The calibration curve was linear over the established range with R2 > 0.997. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01-15.20 μg/mL and 0.03-46.06 μg/mL. The method exhibited an intraday and interday precision range of 96.25-105.66% and 93.52-109.92%, respectively (RSD <2.80%). The recoveries and relative standard deviation (RSD) of the four compounds in Tilia amurensis were in the range of 90.42-104.84% and 0.2-2.58%.Conclusion:This developed method was accurate and reliable for the quality evaluation of the four compounds isolated from Tilia amurensis.
    Pharmacognosy Magazine 07/2014; 10(39):195-9. DOI:10.4103/0973-1296.137353 · 1.26 Impact Factor
    • "All validation was showed for determination of analysis sample in this study. Linearity, limit of detection (LOD), limit of quantification (LOQ), precision, and recovery were evaluated for the method.[12131415] "
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    ABSTRACT: Background:Pyeongwee-San (PWS) has been widely used for treating acute gastritis, chronic, and gastritis.Objective:In this paper, simultaneous determination of five compounds (naringin, hesperidin, glycyrrhizin, atractylenolide III, and magnolol) from traditional medicine PWS using the high performance liquid chromatography (HPLC) was established for quality control.Materials and Methods:Optimum separations were obtained with a SHISEIDO C18 reverse-phase column by gradient elution with 0.1% Trifluoroacetic acid (TFA) water-acetonitrile as the mobile phase. The flow rate was 1 mL/min and detection wavelength was set at 205 nm and 250 nm. Validation of the analytical method was evaluated by linearity, precision, and accuracy test.Results:The calibration curves were linear over the established range with R2 > 0.9978. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.09 to 0.43 and 0.27 to 1.29 μg/mL. The method exhibited intra-day and inter-day precision range between 0.01-1.86% and 0.04-0.35% respectively. The recoveries of five compounds in PWS were in the range between 93.18-106.40%, and 0.20-1.51%. The application of this method was identified through the successful analysis of five compounds in 12 batches of PWS. In addition, identification of five compounds was confirmed by a liquid chromatography method and mass spectrometry.Conclusion:The HPLC method was could be accomplished to the quality control and stable experiment for the preparations consisted of five major compounds.
    Pharmacognosy Magazine 03/2014; 10(Suppl 1):S22-9. DOI:10.4103/0973-1296.127335 · 1.26 Impact Factor
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    • "Previously, our laboratory has screened the CYP3A substrates from a large number of Traditional Chinese Medicines that are classified as superior drugs (Wu et al., 2012). Fructus Schisandrae has been used for thousands of years, owing to its diverse pharmacological effects (Wei et al., 2010). Many structurally similar dibenzocyclooctadiene lignans from Fructus Schisandrae were found to be metabolized by CYP3A4. "
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    ABSTRACT: To accurately predict the modifications done during metabolic processes by Cytochrome P450 (CYP) 3A enzyme, selecting substrates that best represent a broad range of substrate substitutions and that follow the Michaelis-Menten kinetic properties is highly necessary. In the present study, the oxidative pathways of deoxyschizandrin (DS), the most abundant lignan in Fructus Schisandrae fruit extract, were characterized with liver microsomes from human (HLM) and rat (RLM). Only one mono-hydroxylated metabolite 7(S)-hydroxylated metabolite (isoschizandrin, ISZ) was identified using LC-MS and NMR techniques. CYP3A4 and CYP3A5 were found to be the major isoforms involved in the mono-hydroxylation of DS. Also, the kinetic studies showed that DS hydroxylation obeyed the Michaelis-Menten kinetics both in HLM and in RLM. However, the subsequent metabolism of ISZ was nearly non-existent when DS was present. More importantly, the interactions between DS and three well-characterized CYP3A probe substrates, testosterone (TST), midazolam (MDZ), and nifedipine (NIF), were studied. TST and MDZ were shown to compete with DS for the mutual binding site, causing Km to be increased. The presence of DS also lowered the binding affinities for MDZ and TST. However, DS showed only slight inhibitory effects on nifedipine (NIF) oxidation even though NIF was able to inhibit DS hydroxylation in a noncompetitive fashion. These results show that DS is a good representative substrate of MDZ and TST primarily due to their shared, large binding regions on CYP3A. Therefore, DS is an attractive candidate as a novel CYP3A probe substrate for predicting the metabolic modifications in CYP3A activity.
    Drug metabolism and disposition: the biological fate of chemicals 10/2013; 42(1). DOI:10.1124/dmd.113.053884 · 3.25 Impact Factor
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