High-performance liquid chromatographic assay for the determination of Aloe Emodin in mouse plasma

doi:10.1016/j.jchromb.2003.08.012

Journal of Chromatography B

Volume 796, Issue 1, 25 October 2003, Pages 113-119

https://doi.org/10.1016/j.jchromb.2003.08.012 Get rights and content

Abstract

An isocratic high-performance liquid chromatography (HPLC) method was developed and validated to determine Aloe Emodin (AE) in mouse plasma. The analysis required 0.3 ml of plasma and involves extraction with dichloromethane. The HPLC separation was carried out on Symmetry Shield RP18, a mobile phase of methanol–water–acetic acid (65:35:0.2) and fluorescence detection at λ_ex=410 nm and λ_em=510 nm. The retention time of AE was 11.7 min. The assay was linear from 10 to 1000 ng/ml (r²≥0.999), showed intra- and inter-day precision within 7.8 and 4.7%, and accuracy of 87.3–105.7%. Detection limit (LOD) and quantification limit (LOQ) were 4.5 and 5 ng/ml, respectively. The method was applied to determine for the first time the pharmacokinetic of AE in mice.

Introduction

Aloe Emodin (AE) is an hydroxy-anthraquinone (Fig. 1) naturally present in the leaves of some species of Aloe that has shown antitumor and antiproliferative activity in neuroectodermic tumors in vitro and in vivo [1], [2], [3]. The growth of this tumor transplanted in mice with severe combined immunodeficiency SCID was highly inhibited by AE administered as a suspension in DMSO and saline solution (1:25). The findings that AE possesses a selective activity against neuroectodermic tumors and particularly against neuroblastoma, that is a tumor exhibiting low susceptibility to the available drugs is of potential clinical interest. For this reason it is important to perform preclinical pharmacokinetic studies of AE that can optimize the dosage-schedule to be investigated. In addition it would be of interest to evaluate whether the selectivity of action is related to an enhanced distribution of AE in neuroectodermic tumors.

The need of studies on the distribution of AE prompted us to developing an analytical assay for the determination of this agent in biological fluids and tissues.

As far as we know, only method on the isolation, identification and purification of AE from natural extract were published in literature [4], [5].

This paper reports a novel method to extract and analyze AE by high-performance liquid chromatography (HPLC) that has been applied to investigate the pharmacokinetic proprieties of this compound in mice.

Section snippets

Chemicals

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Aloe Emodin (95% purity) was obtained from Sigma–Aldrich, St. Louis, MO, USA (lot #119H1333).
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Control mice plasma was obtained from IFFA, Credo, France.
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Methanol of HPLC grade was obtained from J.T. Baker B.V., Deventer, Holland.
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Acetic acid glacial for analysis was obtained from Carlo Erba, Milan, Italy.
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Dichloromethane for analysis was obtained from Merck, Darmstadt, Germany.
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Water of HPLC grade was obtained from Milli Ro 60 Water System, Millipore, Milford, MA, USA.
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PEG300 was obtained from Fluka

Chromatography

Fig. 2A shows a typical chromatogram of extracted control mouse plasma only treated with the vehicle PEG300. No endogenous interfering substances of plasma were present at the retention time of AE.

Fig. 2B shows a chromatogram of a plasma sample containing AE at the concentration of 5 ng/ml (i.e. LOQ). Significant differences in the peak area of AE were obtained between the responses for the blank and the LOQ sample in the five replicates injected.

Fig. 2C shows a chromatogram of plasma sample

Conclusion

This report describes the procedure to measure AE in mouse plasma. The method shows great linearity and has a high degree of selectivity, sensitivity and good precision and accuracy.

It should be noted that the method is very rapid requiring simple dichloromethane extraction procedure and HPLC analyses of only 14 min run time.

The method is suitable for use in determining plasma and tissue levels in preclinical investigations and will prove useful for evaluating the pharmacokinetic properties of

References (5)

H.Z. Lee et al.
Eur. J. Pharmacol.
(2001)
T. Pecere et al.
Cancer Res.
(2000)

There are more references available in the full text version of this article.

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