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Abstract AI-Helper 아이콘AI-Helper

A rapid, sensitive and selective liquid chromatography-tandem mass spectrometric (LC-MS/ MS) method for the determination of lithospermic acid B (LSB) in rat serum was developed. LSB and internal standard, 7-hydroxy-3-phenyl-chromen-4-one (HPC) were extracted from rat serum with methyl-tert-butyl et...

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제안 방법

  • The molecular ions of LSB and HPC were fragmented at collision energy of 20 eV and 35 eV using argon as collision gas. Detection of the ions was performed by monitoring the transitions of m/z 717 to m/z 519 for LSB and m/z 237 to m/z 117 for HPC. Peak areas for all components were automatically integrated using MassLynx Version 3.
  • complete the method validation. In each batch, QC samples at 15.0, 75.0, and 400 ng/mL were assayed in sets of six replicates to evaluate the intra- and inter-day precision and accuracy. The percentage deviation of the mean from tme values, expressed as relative error (RE), and the coefficient of variation (CV) serve as the measure of accuracy and precision.
  • The absolute recoveries of LSB were determined by comparing the peak area of six extracted samples at the concentrations of 15.0, 75.0, and 400 ng/mL with the mean peak area of recovery standards. Three replicates of each of the recovery standards were prepared by adding LSB and internal standard to blank rat serum extracts.
  • A sensitive, simple, fast and reliable bioanalytical method is required in order to evaluate the pharmacokinetic disposition of LSB. The purpose of this paper was to describe and validate a LC-MS/MS method using liquid-liquid extraction for the determination of LSB in rat serum. The pharmacokinetics of LSB in male Sprague-Dawley rats were also reported.
  • The fragment ions at m/z 519 [the loss of 1-carboxy-2-(3, 4dihydroxy-phenyl)-ethoxy group from [M-H]- ion] and m/z 117 (the loss of 7-hydroxyphenyl and ketone group from [M-H]' ion) were produced as the prominent product ions for LSB and HPC, respectively. The quantification of the analytes was performed using the MRM mode due to the high selectivity and sensitivity of MRM data acquisitions, where the precursor and product ions are monitored. Two pairs of MRM transitions were selected: 717→519 for LSB and /n/2 237117 for HPC (internal standard).
  • 0, and 400 ng/mL with the mean peak area of recovery standards. Three replicates of each of the recovery standards were prepared by adding LSB and internal standard to blank rat serum extracts.
  • After a 1-day recovery period, LSB dissolved in saline was injected intravenously at a dose of 10 ㎎/㎏ into the left femoral vein of the rats (n=4). Venous blood samples sere collected at 2, 5, 10, 15, 30 min, and 1, 2, 4, 6, 8, and 10 h after i.v. injection. Blood samples were immediately centrifuged at 3000×g for 5 min and harvested serum samples were stored at -20℃ until analysis.

대상 데이터

  • The extraction recoveries of LSB from spiked rat serum were determined at the concentrations of 15.0, 75.0 and 400 ng/mL in six replicates. The recoveries of LSB ranged from 73.
  • 0 and 400 ng/mL in six replicates. The recoveries of LSB ranged from 73.2 to 79.5%, with that of HPC (internal standard) being 75.1 ±6.0% (Table Ⅲ). The one-step liquid-liquid extraction with methyl-fert-butyl-ether at pH 2 has been successfully applied to the extraction of LSB from rat serum.
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참고문헌 (9)

  1. Abd-Elazem, I. S., Chen, H. S., Bates, R. B., and Huang, R. C., Isolation of two highly potent and non-toxic inhibitors of human immunodeficiencyvirus type 1 (HIV-1) integrase from Salviami/tiorrhiza. Antiviral Res; 55, 91-106 (2002) 

  2. Hase, K., Kasimu, R., Basnet, P., Kadota, S., and Namba, T., Preventive effect of lithospermate B from Salvia mi/tiorhiza on experimental hepatitis induced by carbon tetrachloride or D-galactosamine/lipopolysaccharide. Planta Med., 63, pp. 2226 (1997) 

  3. Kamata, K., lizuka, T., Nagai, M., and Kasuya, Y., Endotheliumdependent vasodilator effects of the extract from Salvia miltionhizae radix. A study on the identification of Iithospermic acid B in the extracts. General Pharmacol., 24, 977-981 (1993) 

  4. Kamata, K., Noguchi, M., and Nagai, M., Hypotensiveeffects of lithospermic acid B isolated from the extract of Salvia mi/tiorrhiza Radix in the rat. General Pharmacol., 25, 69-73 (1994) 

  5. Kang, D. G., Oh, H., Sohn, E. J., Hur, T. Y., Lee, K. C., Kim, K. J., Kim, T. Y., and Lee, H. S., Lithospermic acid B isolated from Salvia miltiorrhiza ameliorates ischemia/reperfusioninduced renal injury in rats. Life Sci., 75, 1801-1816 (2004) 

  6. Shigematsu, T., Tajima,S., Nishikawa, T., Murad, S., Pinnell, S. R., and Nishioka, I., Inhibition of collagen hydroxylation by lithospermic acid B magnesium salt, a novel compound isolated from Salvia miltiorrhizae Radix. Biochem. Biophys. Acta, 1200, 79-83 (1994) 

  7. Yokozawa,T., Chung, H. Y., Lee, T. w., Oura, H., Nonaka, G., and Nishioka, I., Potentiating effect of converting enzyme inhibitor captopril to the renal responses of magnesium Iithospermate B in rats with adenine-induced renal failure. Chem. Pharm. Bull., 39, 732-736 (1991) 

  8. Yokozawa, T., Dong, E., Liu, Z.W., Shibata, T., Hasegawa, M., Watanabe, H., and Oura, H., Magnesium Iithospermate B ameliorates cephaloridine-induced renal injury. Exp. Toxicol. Pathol., 49, 337-341 (1997) 

  9. Zupko, I., Hohmann, J., Redei, D., Falkay, G., Janicsak, G., and Mathe, I., Antioxidant activity of leaves of Salvia species in enzyme-dependent and enzyme-independent systems of lipid peroxidation and their phenolic constituents. Planta Med., 67, 366-368 (2001) 

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