V Dinesh Kumar, Verma Priya Ranjan Prasad, Singh Sandeep Kumar, Viswanathan S
Department of Pharmaceutical Sciences and Technology, Birla Institute of Technology, Mesra, Ranchi, Jharkhand, India.
Micro-Therapeutic Research Labs Private Limited, Chennai, 600097, India.
Biomed Chromatogr. 2015 Nov;29(11):1731-6. doi: 10.1002/bmc.3486. Epub 2015 May 14.
A simple, rapid and sensitive LC-MS/MS method was developed and validated for the determination of free quercetin in rat plasma, using fisetin as internal standard. The detection was performed by negative ion electrospray ionization under selected reaction monitoring. Chromatographic separation (isocratic elution) was carried out using acetonitrile-10 m m ammonium formate (80:20, v/v) with 0.1% v/v formic acid. The lower limit of quantification (4.928 ng/mL) provided high sensitivity for the detection of quercetin in rat plasma. The linearity range was from 5 to 2000 ng/mL. Intra- and inter-day variability (RSD) of quercetin extraction from rat plasma was <4.19 and 1.37% with accuracies of 98.77 and 99.67%. The method developed was successfully applied for estimating free quercetin in rat plasma, after oral administration of quercetin-loaded biodegradable nanoparticles (QLN) and quercetin suspension. QLN (C(max), 1277.34 ± 216.67 ng/mL; AUC, 17,458.25 ± 3152.95 ng hr/mL) showed a 5.38-fold increase in relative bioavailability as compared with quercetin suspension (C(max), 369.2 ± 108.07 ng/mL; AUC, 3276.92 ± 396.67 ng hr/mL).
建立了一种简单、快速且灵敏的液相色谱-串联质谱法（LC-MS/MS），以非瑟酮为内标物，用于测定大鼠血浆中的游离槲皮素。检测采用负离子电喷雾电离，在选择反应监测模式下进行。色谱分离（等度洗脱）采用乙腈-10 mM甲酸铵（80:20，v/v）并添加0.1% v/v甲酸。定量下限（4.928 ng/mL）为大鼠血浆中槲皮素的检测提供了高灵敏度。线性范围为5至2000 ng/mL。从大鼠血浆中提取槲皮素的日内和日间变异系数（RSD）分别<4.19%和1.37%，准确度分别为98.77%和99.67%。所建立的方法成功应用于口服槲皮素负载的可生物降解纳米颗粒（QLN）和槲皮素混悬液后大鼠血浆中游离槲皮素的估算。与槲皮素混悬液（C(max)，369.2±108.07 ng/mL；AUC，3276.92±396.67 ng·hr/mL）相比，QLN（C(max)，1277.34±216.67 ng/mL；AUC，17458.25±3152.95 ng·hr/mL）的相对生物利用度提高了5.38倍。
