Abstract
Aim:
To develop and validate a novel precolumn derivatization method for the quantitative determination and pharmacokinetic application of acetylshikonin in macaque monkeys by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS).
Methods:
2-Mercaptoethanol was added to the blood sample as the derivatization reagent. The derivatization reaction formed 1 major derivation product, which was well correlated with acetylshikonin. The acetylshikonin concentrations in the biological samples were calculated by quantitative determination of the major derivation product using LC-ESI-MS/MS. Separation was achieved using a C18 column (2 mm×50 mm, 5 μm) at room temperature and a linear gradient elution with a mobile phase containing methanol (1.96% acetic acid) and 10% methanol in water (1.96% acetic acid and 10 mmol/L ammonium acetate) at a flow rate of 0.2 mL/min. In addition, the major derivative, named derivative III, was identified by UV spectra, MS, and the 1H-NMR and 13C-NMR spectra.
Results:
Good linearity was obtained within the range of 5 and 2000 ng/mL (r>0.99 using a linear regression model with 1/x2 weighting) for acetylshikonin. The interday and intraday precisions were found to be less than 12.3%, with the exception of the lowest concentration, which was less than 17.2%. The interday and intraday accuracies, which were between –3% and 0.6%, were also observed. After the administration of acetylshikonin (80 mg/kg, po) in macaque monkeys, the pharmacokinetic parameters were obtained through the non-compartmental analysis, where the area under the concentration-time curve to the last measurable concentration, the terminal elimination half-life, and the mean residual time were 615.4±206.5 ng h/mL,12.3±1.6 h, and 10.2±0.7 h, respectively.
Conclusion:
The method was validated and applied to the quantitative determination and pharmacokinetic study of acetylshikonin in the blood samples of macaque monkeys.
Similar content being viewed by others
Article PDF
References
Li HY, Luo SR, Zhou TH . Studies on in vitro metabolism of shikonin. Phytother Res 1999; 13: 236–8.
Kundakovic T, Fokialakis N, Dobric S, Pratsinis H, Kletsas D, Kovacevic N, et al. Evaluation of the anti-inflammatory and cytotoxic activities of naphthazarine derivatives from Onosma leptantha. Phytomedicine 2006; 13: 290–4.
Singh B, Sharma MK, Meghwal PR, Sahu PM, Singh S . Anti-inflammatory activity of shikonin derivatives from Arnebia hispidissima. Phytomedicine 2003; 10: 375–80.
Papageorgiou VP, Assimopoulou AN, Couladouros EA, Hepworth D, Nicolaou KC . The chemistry and biology of alkannin, shikonin, and related naphthazarin natural products. Angew Chem Int Ed 1999; 38: 270–300.
Huang ZS, Zhang M, Ma L, Gu LQ . A survey of chemical and pharmacologic studies on ZiCao. Nat Prod Res Dev 1999; 12: 73–82 (in Chinese).
Lu Q, Liu WJ, Ding J, Cai JC, Duan WH . Shikonin derivatives: synthesis and inhibition of human telomerase. Bioorg Med Chem Lett 2002; 12: 1375–8.
Papageorgiou VP, Assimopoulou AN, Samanidou VF, Papadoyannis IN . Recent advances in chemistry, biology and biotechnology of alkannins and shikonins. Curr Organ Chem 2006; 10: 2123–42.
Duan ZF, Huang ZS, Bu XZ, Xie WL, Gu LQ . Study on the reaction of natural naphthazarin compounds with thiol nucleophiles. Chin J Organ Chem 2001; 21: 1112–6 (in Chinese).
Duan ZF, Huang ZS, Ma L, Che ZT, Gu LQ . Covalent modification of lactate dehydrogenase and alcohol dehydrogenase by alkannin derivatives. Chin J Organ Chem 2001; 21: 668–71 (in Chinese).
Huang ZS, Gu LQ, Che ZT, Zhang M, Qiu GQ . The nucleophilic reaction of β-dimethylacrylalkannin with aniline and thiophenol. Acta Chim Sin 2000; 58: 1043–9 (in Chinese).
Pang ZG, Wang BQ, Li HF . Study on the compound pharmacokinetics of β-shikonin. J Xi-an Med Univ 2001; 22: 319–20 (in Chinese).
Author information
Authors and Affiliations
Corresponding author
Additional information
This work was supported by Beijing Jin BenCao Chinese Herbal Medicine Technology Development.
Rights and permissions
About this article
Cite this article
Sun, Dx., Tian, Hf., Meng, Zy. et al. Quantitative determination of acetylshikonin in macaque monkey blood by LC-ESI-MS/MS after precolumn derivatization with 2-mercaptoethanol and its application in pharmacokinetic study. Acta Pharmacol Sin 29, 1499–1506 (2008). https://doi.org/10.1111/j.1745-7254.2008.00898.x
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1111/j.1745-7254.2008.00898.x