A rapid HPLC/ESI-MS/MS method for quantitative analysis of isovalerylshikonin in rat plasma.
A new, fast and sensitive high-performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) method was developed and validated for isovalerylshikonin in rat plasma using emodin as internal standard (IS). The analyte was extracted from rat plasma with ethyl acetate, after 10% HCl treatment and protein precipitated by methanol. The compound was separated on an Ultimate XB-C(18) analytical column using a mobile phase of methanol-10 mM ammonium acetate in water-acetonitrile containing 0.05% formic acid (45 : 10 : 45, v/v/v) with isogradient elution. The analyte was detected in negative ion mode using multiple-reaction monitoring. The method was validated and the specificity, linearity, lower limit of quantitation (LLOQ), precision, accuracy, recoveries and stability were determined. LLOQ was 9 ng/mL for isovalerylshikonin. Correlation coefficient (r) value for the linear range of the analyte was greater than 0.99. The intra-day and inter-day precision and accuracy were better than 8.52%. The relative and absolute recovery was above 86% and no matrix effects were observed for isovalerylshikonin. This validated method provides a modern, rapid and robust procedure for the pharmacokinetic study of the two compounds in rats after intravenous administration to rats (n = 4).
Zhu
MeiXiang
M
Department of Chemistry and Key Laboratory for Chemical Biology of Fujian Province, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, People's Republic of China.
Gao
Yanbing
Y
Wu
Zhen
Z
Zhao
YuFen
Y
D000818:Animals; D028082:Boraginaceae / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D004365:Drugs, Chinese Herbal / Q000032:analysis* / Q000737:chemistry / Q000493:pharmacokinetics; D004642:Emodin / Q000737:chemistry; D007275:Injections, Intravenous; D009285:Naphthoquinones / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000493:pharmacokinetics; D010421:Pentanoic Acids / Q000097:blood* / Q000737:chemistry; D018517:Plant Roots / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
D004365:Drugs, Chinese Herbal; D009285:Naphthoquinones; D010421:Pentanoic Acids; C550538:isovalerylshikonin; D004642:Emodin