A method of quantitative determination of camphecene, a new anti-influenza agent, in rat blood plasma based on LC-MS/MS was developed, validated and used to study the distribution of the agent between blood cells and blood plasma. The method was validated according to FDA and EMA recommendations in terms of selectivity, linearity, accuracy, precision, recovery, stability and carry-over. Plasma samples were precipitated with methanol followed by the addition of a methanolic solution of 2-adamantylamine hydrochloride (internal standard). HPLC analysis was performed on a reversed-phase column; the total time of analysis was 11 min, including column equilibration. MS/MS detection was performed on a 3200 QTRAP triple quadrupole mass spectrometer in multiple reaction monitoring (MRM) mode. Transitions 196.4 → 122.2/153.3 and 152.2 → 93.1/107.2 were monitored for camphecene and the internal standard, respectively. The calibration curve was built in the concentration range of 10-5000 ng/ml. The intra-day and inter-day accuracy and precision, carry-over and recovery were within the acceptable limits. It was found that, after spiking blood with camphecene and separating plasma, the concentration of the substance in the latter was close to its initial concentration in the blood. This property of the substance may be useful for clinical trials of the agent. It has also been established that the process of camphecene distribution (adsorption) between blood cells and blood plasma is reversible, and the amount of adsorbed substance is linearly dependent on its initial concentration in the blood for a wide range of concentrations, temperatures and hematocrit values.

中文翻译：

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LC-MS / MS方法在大鼠血浆中抗流感药物樟脑的定量分析方法的开发和验证，及其在研究该药物在血浆中的分布方面的应用。

建立了一种基于LC-MS / MS的新型抗流行病剂樟脑烯定量测定方法，并进行了验证，并用于研究该剂在血细胞和血浆之间的分布。根据FDA和EMA的建议，该方法在选择性，线性，准确度，精密度，回收率，稳定性和残留方面得到了验证。用甲醇沉淀血浆样品，然后加入2-金刚烷基胺盐酸盐的甲醇溶液（内标）。HPLC分析是在反相柱上进行的。分析的总时间为11分钟，包括色谱柱平衡。MS / MS检测是在3200 QTRAP三重四极杆质谱仪上以多反应监测（MRM）模式进行的。过渡196.4→122.2 / 153.3和152.2→93.1 / 107。分别对2个进行了二十碳烯和内标物的监测。校准曲线建立在10-5000 ng / ml的浓度范围内。日内和日间准确性和精密度，结转和恢复均在可接受的范围内。结果发现，在用二十碳烯向血液加标并分离血浆后，血浆中该物质的浓度接近其在血液中的初始浓度。物质的这种性质可能对药物的临床试验有用。还已经确定，血细胞和血浆之间的蝶烯分布（吸附）过程是可逆的，并且在很宽的浓度，温度和血细胞比容值范围内，吸附物质的量线性依赖于其在血液中的初始浓度。 。