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目的采用LC-MS/MS法,对于重要药材知母中的主要活性成分之一芒果苷,测定其在大鼠血浆中的浓度,研究芒果苷在正常大鼠及糖尿病大鼠体内的代谢情况。方法仪器采用Thermo公司的TSQ三重四级杆及ESI源,配合WatersAcquity UPLC液相。色谱条件:色谱柱为Athena C18-WP柱(50mm×2.1mm,3μm)。分析流动相为甲醇/水(5:495V/V,0.2%甲酸;A相),甲醇/水(495:5V/V,0.2%甲酸;B相),流速0.35m.lmin-1,柱温为室温。质谱条件:ESI离子源,检测方式为正离子模式([M+H]峰),用于定性分析离子对为芒果苷m/z:423.2～273.2,商陆皂苷(Is)为m/z:849.4～335.1。用于血药提取的方法为乙酸乙酯萃取。糖尿病模型造模方法为:腹腔注射10%四氧嘧啶120 mg·kg-1,24 h后再次注射100mg·kg-1,1 d后及1周后空腹血糖值≥11.1 mmol·L-1为造模成功。造模成功后,健康大鼠和模型组大鼠分别静脉注射芒果苷2 mg·kg-1,另外灌胃10mg·kg-1,进行血药浓度测定。结果方法学验证结果:芒果苷在10.15～1300ng.ml-1的范围内呈线性(R2=0.9932),日内和日间精密度小于14.6%,日内和日间准确度在95.3%～114.3%范围内。芒果苷的基质效应在120.7%～131.1%范围内。PK测定的结果为健康大鼠药物口服后的达峰时间约为给药后1h,口服生物利用度为1.7%,清除率为11.4m.lh-1.kg-1,分布体积为88.3ml.kg-1。静脉注射后半衰期为6 h左右。糖尿病模型大鼠达峰时间基本不变,但口服生物利用度明显下降,半衰期延长。结论芒果苷口服后吸收迅速,但口服生物利用度较低;其脂溶性、分子内可旋转键的数量、分子的极性表面积等应属于易于吸收的分子,但其又为P糖蛋白的底物,并且有报道肠道菌群对其有降解作用,因此口服时导致较大的首过效应。糖尿病模型大鼠口服生物利用度明显下降,可能与疾病造成动物体内微环境的改变,从而改变了机体吸收药物的能力。
Objective To determine the concentration of mangiferin in rat plasma by LC-MS / MS method and to study the metabolism of mangiferin in normal rats and diabetic rats. Methods The instrument was equipped with a Thermo TSQ triple quadrupole and ESI source for use with a Waters Acquity UPLC liquid phase. Chromatographic conditions: Athena C18-WP column (50 mm × 2.1 mm, 3 μm). The mobile phase consisted of methanol / water (5: 495 V / V, 0.2% formic acid; Phase A), methanol / water For room temperature. Mass spectrometry conditions: ESI source, the detection mode is positive ion mode ([M + H] peak), for qualitative analysis of the ion pair of mangiferin m / z: 423.2 ~ 273.2, 849.4 ~ 335.1. The method used for blood drug extraction is ethyl acetate extraction. The model of diabetes mellitus was established by intraperitoneal injection of 10% alloxan 120 mg · kg-1,24 h after re-injection of 100 mg · kg -1,1 d and 1 week after fasting blood glucose ≥11.1 mmol·L-1 Modeling success. After successful modeling, the rats in healthy group and model group were given intravenous injection of mangiferin 2 mg · kg-1, respectively, and the other 10 mg · kg-1 orally. Results Methodological validation results showed that mangiferin showed linearity (R2 = 0.9932) in the range of 10.15 ~ 1300 ng · ml-1, the intra- and inter-day precision was less than 14.6%, the intra- and inter-day accuracy was in the range of 95.3% -114.3% Inside. Matrix effects of mangiferin ranged from 120.7% to 131.1%. The result of PK measurement shows that the peak time after oral administration of the medicine for healthy rats is about 1 hour after administration, the oral bioavailability is 1.7%, the clearance rate is 11.4m.lh-1.kg-1, and the distribution volume is 88.3ml. kg-1. The half-life after intravenous injection is about 6 h. Diabetes model rats peak time unchanged, but the oral bioavailability decreased significantly, the half-life prolonged. Conclusion Mangiferin absorbed rapidly after oral administration, but its oral bioavailability is low. Its lipophilicity, the number of intramolecular rotatory bonds and the polar surface area of the molecule should be easily absorbed molecules, but it is also the end of P glycoprotein It has been reported that gut microbiota has a degrading effect on it and therefore leads to a large first pass effect when administered orally. Oral bioavailability of diabetic rats significantly decreased, which may cause changes in the microenvironment of the animal, thereby changing the ability of the body to absorb drugs.