δ-viniferin（Rs-1）是从葡萄中分离得到的天然产物，大量研究发现，Rs-1有抗炎，抗微生物，抗氧化，抗糖尿病和心血管保护等多种药理活性，AcRs-1为将Rs-1的5个酚羟基都乙酰化的衍生物。本研究比较了口服或静脉Rs-1和AcRs-1的药代动力学性质。

本文建立了LC-MS/MS分析方法对大鼠血浆、尿液及粪便中的Rs-1和AcRs-1进行了测定，并测定了它们的代谢产物。生物样品（血浆、尿液和粪便）前处理采用乙酸乙酯或乙醚提取，40ºC氮气吹干，甲醇或乙腈复溶进样；色谱条件为：Shim-pack XR-ODS C18色谱柱（3×75 mm，2.2 μm），流动相为0.1%甲酸∶0.1%甲酸乙腈梯度洗；质谱条件为：采用电喷雾离子源（ESI），多反应离子检测（MRM）进行扫描。Rs-1选用橙皮素为内标（IS-1），负离子模式，定量离子对为：m/z 453.1/411.1（Rs-1）和m/z 301.0/164.0（IS-1）。AcRs-1选用五味子甲素为内标（IS-2），正离子模式，定量离子对为m/z 665.3/581.1（AcRs-1）和m/z 417.1/316.1（IS-2）。

Rs-1在血浆、尿液和粪便中线性范围为1.42~2172ng/mL，最低定量限为1.42 ng/mL，相关系数r>0.99，日内、日间精密度RSD<3.7％，准确度在−4.9~5.6%之间；提取回收率在70.7% ~ 90.4%间；基质效应在85.8%~100.4%。方法快速、灵敏、重现性好。AcRs-1在血浆、尿液和粪便中线性范围为1.33~2022ng/mL，最低定量限为1.33 ng/mL，相关系数r>0.99，日内、日间精密度RSD<4.3％，准确度在−3.6~2.8%之间；提取回收率在82.2%~98.2%间；基质效应在90.1%~100.7%。方法快速、灵敏、重现性好。

采用所建立的生物样品前处理及LC-MS/MS测定方法，研究了大鼠等摩尔量口服灌胃（155 μmol/mL）或静脉注射（3 μmol/mL）Rs-1和AcRs-1后的药代动力学特征，采用DAS软件计算血浆药时曲线与药动学参数。

研究了Rs-1和AcRs-1大鼠静脉注射给药的药代动力学。两者血浆中均发现少量代谢产物Rs-1葡萄糖醛酸结合物和硫酸酯结合物。Rs-1静脉注射以其原型药物为主。由于体内含有磷酸酯酶，AcRs-1进入体内后酯键水解脱去乙酰基，可以代谢为多种失去不同数量乙酰基的化合物，主要为Rs-1。AcRs-1静脉注射以AcRs-1及其去乙酰化的代谢产物总和计算，其清除率较Rs-1静脉注射低。

研究了Rs-1和AcRs-1大鼠口服给药给药的药代动力学。口服Rs-1后，Rs-1在体内迅速代谢成Rs-1葡萄糖醛酸结合物和硫酸酯结合物，原型药物在体内浓度较低，绝对生物利用度仅为2.3%，加上两种代谢产物计算吸收率可以达到31.5%。排泄实验中表明，口服Rs-1后60小时之内，只有0.09%的药物从尿液中排出，其中包括Rs-1原型和其两种代谢产物；60.3% 以原型形式从粪便中排出。口服AcRs-1后，血液里检测不到原型药物AcRs-1和其去乙酰化的代谢产物以及Rs-1葡萄糖醛酸结合物和硫酸酯结合物。而排泄实验发现，尿中也未检测到原型药和代谢产物，粪便中检测到了原型药AcRs-1以及失去一个到五个不等乙酰基的代谢产物，其总和占口服剂量的94.8%，其中原型药物AcRs-1占78.3%。

本实验对Rs-1和AcRs-1进行了早期药代动力学研究，测定了在大鼠体内的生物利用度及排泄，阐明其体内药动学特征。结果表明，AcRs-1较Rs-1生物利用度更低，可能由于其分子量过大和溶解性更差导致其更难以吸收。本研究为评估成药性提供了科学依据，并为接下来的结构改造提供了重要参考和研究思路。

Trans δ-viniferin (Rs-1) was found in grapevine (Vitis vinifera L.), and many studies have demonstrated that Rs-1 has lots of pharmacological abilities, such as anti-inflammation, antimicrobial effect, antioxidation, cardioprotection and antidiabetes. Penta-O-acetyl-δ-viniferin (AcRs-1) was synthesized by acetylating five hydroxyl groups of δ-viniferin. It has higher hydrophobicity than the parent molecule, which might increase the ability to cross the cellular membrane. In this study, we investigate and compare the pharmacokinetics of Rs-1 and AcRs-1 in rats following a single oral administration or intravenous injection.

In the present study, a sensitive and accurate LC–MS/MS method was developed and validated for the determination of Rs-1 and AcRs-1 in rat plasma, urine and feces. A liquid-liquid extraction (LLE) method was applied to extract the analyte from biological samples (plasma, urine and feces), and the extraction solvent was ethyl acetate or diethyl ether. The supernate was evaporated to dryness under nitrogen at 40°C. Then the residue was reconstituted in methanol or acetonitrile. Chromatographic separation was achieved on a shim-pack XR-ODS column (3×75 mm, 2.2 μm) using gradient mobile phase with acetonitrile (0.1% formic acid)–water (0.1% formic acid). hesperetin was used as an internal standard (IS-1) of Rs-1, and MS/MS detection was performed by negative ion electrospray ionization using MRM transitions at m/z 453.2→411.2 for Rs-1 and m/z 301.1→163.9 for IS-1. Deoxyschizandrin was used as an internal standard (IS-2) of AcRs-1, and MS/MS detection was performed by positive ion electrospray ionization using MRM transitions at m/z 665.1→581.2 for Rs-1 and m/z 417.2→316.2 for IS-2.

The method of Rs-1 was sensitive with a lower limit of quantification of 1.42 ng/mL and linear over the range of 1.42 ~ 2172 ng/mL in all matrices, r > 0.99. The intra-day and inter-day precision (RSD, %) values were both ≤ 3.7%, the accuracy (RE, %) were in the range of –4.9 to 5.6%. The extraction recovery was 70.7% ~ 90.4%, RSD < 9.3％. The matrix effect was 85.8% ~ 100.4%, RSD < 4.7%。

The method of Rs-1 was sensitive with a lower limit of quantification of 1.33 ng/mL and linear over the range of 1.33 ~ 2022 ng/mL in all matrices, r > 0.99. The intra-day and inter-day precision (RSD, %) values were both ≤ 4.3%, the accuracy (RE, %) were in the range of −3.6 ~ 2.8%%. The extraction recovery was 82.2% ~ 98.2%%. The matrix effect was 90.1% ~ 100.7%。

The reliable method was subsequently applied to the assessment of the pharmacokinetics, bioavailability, metabolism and excretion of AcRs-1 and Rs-1 in healthy rats following a single oral administration (155 μmol/ml) or intravenous injection (3 μmol/ml). The pharmacokinetic parameters were evaluated by non-compartmental model using DAS Software. The results were as follows:

After intravenous injection of Rs-1 or AcRs-1, a small amount of Rs-1 glucuronide and sulfate conjugates were detected. After intravenous injection of Rs-1, Rs-1 was the main compound in the plasma. While owe to the phosphatases, AcRs-1 in plasma can easily lose the acetyls. There were some compounds with three acetyls, two acetyls, one acetyls within and Rs-1 1 h after intravenous injection of AcRs-1, and the t_1/2 of the total Rs-1s (Rs-1 and Rs-1 with diffrent amount of acetyls) has been prolonged and the Area under the curve (AUC) has been enhanced compare to intravenous injection of Rs-1.

After oral administration Rs-1, it was metabolized rapidly to Rs-1 glucuronide and sulfate conjugates. The absolute oral bioavailability of Rs-1 was 2.3%, and the total absorption was rose to 31.5% in addition to its glucuronide and sulfate metabolites. Only 0.09% of the gavage dose, including Rs-1 and it metabolites, were excreted through urine; while 60.3% were through feces in unchanged form. After oral administration of AcRs-1, there was little AcRs-1 or any other form of metabolites in plasma and urine, while 94.8% was recovered in feces (contain Rs-1 and Rs-1 with diffrent amount of acetyls) and mainly remained unchanged, which implied that AcRs-1 was barely absorbed (78.3%).

This study compared pharmacokinetics, bioavailability, metabolism and excretion of AcRs-1 and Rs-1. The results showed that AcRs-1 was barely absorbed after oral administration. Maybe the more poor solubility and high molecular weight makes AcRs-1 was more difficult to absorb. Acetylation of Rs-1 was not a good way to improve the bioavailability. The developed method and the pharmacokinetic data can provide a basis for further studies.