Short communicationDetermination of scutellarin in rat plasma by high-performance liquid chromatography with ultraviolet detection
Introduction
Flavonoids are widely occurring in the plant kingdom and contained in the common human diet, and comprise of flavones, flavonols, flavanones, flavanonols, isoflavone, flavanol, anthocyanidins. Many of these compounds exist as sugar conjugates [1]. Scutellarin, a flavone glucuronide, extracted from a Chinese herb Erigero breviscapus (Vant.) Hand.-Mazz [2], is not only an important component of a Chinese herb, but also a major constituent of Skullcap, a popular western herb. It has been used in the treatment of occlusive cerebral vascular diseases. It is efficacious in the treatment of cerebral infarction, coronary heart disease, angina pectoris [3], [4], [5]. For a better understanding of its pharmacokinetics, it is essential to use a sensitive and precise analytical method to determine the concentration of scutellarin in biological fluids.
There has been active interest in recent years in developing and optimizing analytical methods for detection of flavonoids in foods [6], [7], [8] and biological matrix. A number of methods have been described for flavonoids in biological fluids [9], [10], [11], [12], [13], [14], [15], [16], [17], [18], [19], [20], including the determination of baicalin and baicalein in rat plasma by HPLC with electrochemical detection [9], and the quantitation of wogonin and its major metabolite wogonin-7β-d-glucuronide in rat plasma by LC–MS/MS [10]. Tsuchiya analyzed polyhydroxyflavones in human plasma using HPLC with diode array detection and solid-phase borate-complex extraction [11]. Several recent reports described the analysis of quercetin in human plasma using HPLC with electrochemical detection [12] and the identification of quercetin glucuronides in human plasma by LC–MS/MS [13]. Nielsen et al. determined the apigenin and acacetin in human urine by column-switching HPLC–UV [14]. However, to our knowledge, there is still no method described for the determination of scutellarin in biological fluids.
In the present study, we firstly established an HPLC–UV method that is sensitive, simple and suitable for determining scutellarin in rat plasma.
Section snippets
Materials
Scutellarin was from the Delta Information Center for Natural Organic Compounds (99.5%, Hong Kong, China). Injectio Breviscapini, which is an injection solution of scutellarin (20 mg/5 ml) was produced by Gejiu Bio-Medicine Industry Ltd. (Yunnan, China). 2-(4-Hydroxy-phenyl)-7-(3-morpholin-4-yl-propoxy)-chromen-4-one (internal standard, IS, see Fig. 1) was a kind of gift from Prof. Chun Hu (Shenyang Pharmaceutical University, Shenyang, China). Methanol and acetonitrile were of HPLC-grade, and
Optimization of HPLC conditions
In optimizing the chromatographic conditions, the pH of the mobile phase and organic modifier were explored. The mobile phases of pH 6.5 or 4.0 yielded tailing peaks. When the pH value of the mobile phase decreased from 6.5 to 4.0, the peaks became sharper. The peaks were sharp and symmetric when the pH value of the mobile phase was adjusted to 2.5. The use of an acid modifier is important to suppress ionization of the weak acidic phenolic group and interactions of these groups with residual
Conclusion
In this study, a specific and sensitive assay for the determination of scutellarin in rat plasma was presented. The method is simple, rapid, and applicable to preliminary pharmacokinetic studies of scutellarin in rats.
Acknowledgements
This work was supported by grants 39930180 and 30271525 of the National Natural Science Foundation of China. We are thankful to Professor Chun Hu for the supply of 2-(4-hydroxy-phenyl)-7-(3-morpholin-4-yl-propoxy)-chromen-4-one.
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2017, Asian Journal of Pharmaceutical SciencesSimultaneous determination of scutellarin and tetrahydropalmatine of Deng-yan granule in rat plasma by UFLC-MS/MS and its application to a pharmacokinetic study
2014, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :Pharmacokinetic researches of TCM could explain and predict useful events related to the efficacy and toxicity, and are helpful to evaluate the rationality and compatibility of herbs or prescriptions [18]. Up to now, several analytical methods have been developed to quantify scutellarin and tetrahydropalmatine in biological matrices, including high-performance liquid chromatography with ultraviolet detection (HPLC-UV) [19,20] and liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) [21–23]. However, there is no method established for simultaneous determination of scutellarin and tetrahydropalmatine in vivo.
Simultaneous determination of three glucuronide conjugates of scutellarein in rat plasma by LC-MS/MS for pharmacokinetic study of breviscapine
2014, Journal of Chromatography B: Analytical Technologies in the Biomedical and Life SciencesCitation Excerpt :These activities are mainly attributed to the presence of a large amount (>85%) of scutellarin (scutellarein-7-O-β-d-glucuronide) in breviscapine. It has previously been demonstrated in the literature that plasma concentration of scutellarin was extremely low following oral administration of either breviscapine or scutellarin to mice [6], rats [7,8], rabbits [9], dogs [10] and humans [11,12]. Subsequent investigations revealed that scutellarin underwent extensive first-pass metabolism (hydrolysis to scutellarein) in the intestinal tract before being absorbed into the blood circulation [12–16].
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2008, International Journal of PharmaceuticsCitation Excerpt :Fifty microliters of protocatechualdehyde methanol solution (68.2 μg mL−1) and 100 μL of 1 M phosphoric acid was added to the each blood sample (0.5 mL). Plasma sample was extracted with 3 mL of ethyl acetate after being acidificated by 100 μL of 0.01 M HCl (Dafang Zhong et al., 2003). After the samples were vortexed for 3 min, the mixture was centrifuged (TGL-16C, Anke Co., China) for 10 min at 3000 rpm.
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