离体外翻肠囊法研究白及提取物中6种成分的肠吸收特性

为研究白及提取物中4-(葡萄糖氧基)-肉桂酸葡萄糖氧基苄酯(B12)、2-异丁基苹果酸(B6)、1-[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯(B17)、1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯(B14)、二氢菲1(B19)和1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯-2-[4-O-肉桂酰基-6-O-乙酰基]葡萄糖苷(B23)6个成分在大鼠小肠中的吸收动力学特性。实验采用大鼠离体外翻肠囊模型考察各成分在不同浓度、不同肠段的吸收特性,建立超高效液相色谱-三重四极杆串联质谱联用仪(UPLC-MS/MS)法测定各成分含量,计算累计吸收量和吸收速率常数。结果表明,除B6成分的低浓度外,各成分在不同浓度、不同肠段下均表现为线性吸收,其回归相关系数(R)均达到0.9以上,符合一级吸收速率,且吸收速率常数随着浓度的增加而增加,提示各成分吸收机制为被动吸收;在同一浓度下不同肠段的总体吸收趋势为十二指肠的吸收要大于回肠、结肠和空肠。综上,白及提取物中6种成分在小肠中均有吸收,但小肠对各成分的吸收具有选择性。本研究可为白及提取物的药物临床开发,确定药物剂型方面提供了一定的实验参考。     

UPLC-MS/MS同时测定白及中6个指标成分的含量

建立UPLC-MS/MS同时测定白及药材中的α-异丁基苹果酸(B6)、4-(葡萄糖氧基)-肉桂酸葡萄糖氧基苄酯(B12)、1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯(B14)、1-[4-(葡萄糖氧)苄基]-2-异丁基苹果酸(B17)、二氢菲1(B19)和1,4-二[4-(葡萄糖氧)苄基]-2-异丁基苹果酸酯-2-[4-O-肉桂酰基-6-O-乙酰基]葡萄糖苷(B23)含量的分析方法,采用Acquity UPLC BEH C18柱(2.1 mm×50 mm,1.7μm);流动相0.1%甲酸乙腈溶液(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,采用电喷雾离子源(ESI),多反应离子监测(MRM)模式进行正负离子同步监测。各指标成分之间分离度良好,在选定的浓度范围内线性关系良好(r≥0.9974),平均加样回收率为98.07%~103.19%,RSD3.04%为,重复性、精密度和稳定性良好。此方法操作简便,快速,灵敏度高,可用于白及药材中主要活性成分的含量测定,并为白及药材的质量控制提供了新方法。     

羊耳菊活性部位提取物在大鼠胆汁中代谢产物的研究

采用超高效液相色谱-四极杆串联飞行时间质谱(UHPLC/Q-TOF MS/MS)技术,分析大鼠灌胃羊耳菊活性部位提取物后,主要成分在大鼠胆汁中的排泄方式。SD大鼠胆管插管手术后,灌胃给予羊耳菊活性部位提取物(100 g/kg生药量),收集给药后12 h的胆汁样品。样品经正丁醇液液萃取法处理后,采用UHPLC/Q-TOF MS/MS对胆汁中的主要代谢产物的结构进行分析推测。实验结果显示,胆汁中共检测到27个代谢产物,样品中检测到大量的咖啡酰基奎宁酸还原,甲基化,葡萄糖醛酸化和甲基葡萄糖醛酸化代谢产物,以咖啡酰基奎宁酸的甲基葡萄糖醛酸化为主。说明大鼠口服羊耳菊提取物后,成分可通过胆汁进行消除,并且甲基葡萄糖醛酸结合物是主要的物质形式。     

白皮杉醇苷在大鼠体内外的葡萄糖醛酸结合代谢研究北大核心CSCD

白皮杉醇苷(PG)是藏边大黄中一种天然抗氧化剂,前期研究发现其极易发生代谢。本文主要研究PG在大鼠体内外的葡萄糖醛酸结合代谢特征。SD大鼠经尾静脉注射给予PG(20 mg/kg),采集给药后胆汁样品,采用LC-MS对主要代谢产物进行结构推测。在此基础上,研究大鼠肝微粒体体外温孵体系中PG的葡萄糖醛酸结合代谢,并测定酶促反应动力学参数。实验结果显示SD大鼠经尾静脉注射给予PG,可在胆汁中快速检测到多种PG及其衍生物的葡萄糖醛酸结合代谢产物。在大鼠肝微粒体体外温孵体系中,PG代谢生成两个与体内一致的单葡萄糖醛酸结合代谢物,其葡萄糖醛酸结合代谢的最大反应速率(Vmax)、米氏常数(Km)和肝内清除率CLint(Vmax/Km)分别为10.11 nmol/(min·mg)、0.36 mmol/L和0.028 m L/(min·mg)。PG经静脉途径进入大鼠体内可经肝脏被快速地广泛代谢,葡萄糖醛酸结合代谢是其体内消除的主要途径之一。大鼠肝脏的葡萄糖醛酸转移酶对PG有较强的亲和力,可催化PG发生快速的葡萄糖醛酸结合代谢。     

白皮杉醇苷在大鼠体内外的葡萄糖醛酸结合代谢研究

白皮杉醇苷(PG)是藏边大黄中一种天然抗氧化剂,前期研究发现其极易发生代谢。本文主要研究PG在大鼠体内外的葡萄糖醛酸结合代谢特征。SD大鼠经尾静脉注射给予PG(20 mg/kg),采集给药后胆汁样品,采用LC-MS对主要代谢产物进行结构推测。在此基础上,研究大鼠肝微粒体体外温孵体系中PG的葡萄糖醛酸结合代谢,并测定酶促反应动力学参数。实验结果显示SD大鼠经尾静脉注射给予PG,可在胆汁中快速检测到多种PG及其衍生物的葡萄糖醛酸结合代谢产物。在大鼠肝微粒体体外温孵体系中,PG代谢生成两个与体内一致的单葡萄糖醛酸结合代谢物,其葡萄糖醛酸结合代谢的最大反应速率(Vmax)、米氏常数(Km)和肝内清除率CLint(Vmax/Km)分别为10.11 nmol/(min·mg)、0.36 mmol/L和0.028 m L/(min·mg)。PG经静脉途径进入大鼠体内可经肝脏被快速地广泛代谢,葡萄糖醛酸结合代谢是其体内消除的主要途径之一。大鼠肝脏的葡萄糖醛酸转移酶对PG有较强的亲和力,可催化PG发生快速的葡萄糖醛酸结合代谢。     

糖尿病大鼠肋间肌酶组织化学研究

目的探讨糖尿病早期肋间肌酶组织化学变化。方法应用酶组织化学方法观察糖尿病2周和4周大鼠肋间肌组织脱氢酶、水解酶和氧化酶活性变化。结果糖尿病2周大鼠肋间肌组织琥珀酸脱氢酶、谷氨酸脱氢酶和辅酶Ⅰ黄递酶活性较对照组增强,乳酸脱氢酶活性较对照组减弱,苹果酸脱氢酶、异柠檬酸脱氢酶、葡萄糖-6-磷酸脱氢酶、酸性磷酸酶、酸性-α-萘酸性酯酶和细胞色素氧化酶无变化。糖尿病4周大鼠肋间肌组织琥珀酸脱氢酶、苹果酸脱氢酶、谷氨酸脱氢酶、辅酶Ⅰ黄递酶、酸性磷酸酶和酸性-α-萘酸性酯酶活性较对照组增强,乳酸脱氢酶和细胞色素氧化酶活性较对照组减弱,异柠檬酸脱氢酶、葡萄糖-6-磷酸脱氢酶无变化。结论糖尿病2周大鼠肋间肌组织有氧氧化代谢能力增强,糖酵解能力减弱。糖尿病4周大鼠肋间肌组织有氧氧化能力增强、糖酵解能力减弱及能量代谢紊乱。在糖尿病早期呼吸肌存在代谢异常。     

植物呼吸及代谢的研究 Ⅴ.水稻幼苗亚细胞颗粒的氧化途径

本文报导了关于4天水稻黄化幼苗地上部的亚细胞颗粒氧化丙酮酸的途径的研究。下述结果证明在其中有三羧酸循环运行:1.琥珀酸、α-酮基戊二酸能迅速地被氧化,柠檬酸、苹果酸、延胡索酸以顺序降低的速率为此颗粒制剂氧化。2.丙酮酸的氧化能为催化量的琥珀酸所引发,说明有缩合酶的活性存在。3.琥珀酸的氧化能为丙二酸所抑制。α-酮基戊二酸的氧化能为亚砷酸钠所抑制,并且此被抑制的耗氧可借加入琥珀酸而得到恢复。4.氧化产物的纸上层析鉴定表明:琥珀酸能转化为延胡索酸、苹果酸和异柠檬酸;α-酮基戊二酸能转化为琥珀酸、延胡索酸和苹果酸。对亚细胞制剂及组织匀浆所作异柠檬酸酶及苹果酸合成酶的活性鉴定指出,在水稻幼苗氧化丙酮酸的途径中,乙醛酸循环可能与三羧酸循环同时存在。     

体循环肠灌流法研究红禾麻提取物在类风湿性关节炎大鼠与正常大鼠体内的肠吸收差异

比较红禾麻提取物在AA大鼠与正常大鼠体内的肠吸收差异。实验采用体循环肠灌流法,建立大鼠RA模型,应用UPLC-MS/MS检测肠灌流液中新绿原酸、绿原酸、隐绿原酸、芦丁、异槲皮苷、槲皮苷、山奈酚-3-O-芸香糖苷、木犀草苷的含量,探究各成分在不同因素(pH值、药物浓度、肠段、胆汁和P-gp抑制剂)下的肠吸收特性。结果表明,槲皮苷的吸收方式为主动转运,其余7个成分的吸收方式为被动扩散。除绿原酸、隐绿原酸外,正常状态下各成分以回肠吸收最好,病理状态下以十二指肠吸收最好,推测药物吸收的特定部位会受疾病状态的影响而发生改变。各成分的吸收均受pH、胆汁和P-gp的影响,其中,槲皮苷可能是P-gp的底物。RA能影响红禾麻提取物在大鼠体内的肠吸收,其作用机制尚需进一步研究。     

拟诺卡氏放线菌YIM90087的代谢产物研究北大核心CSCD

对拟诺卡氏菌YIM90087的固体发酵提取物进行了化学成分研究,从中分离得到10个化合物。通过波谱数据分析,鉴定其结构分别为:(3S)-3-苄基-2,5-哌嗪二酮(1)、(3S,6S)-3-(2-丁基)-6-(1-羟乙基)-2,5-哌嗪二酮(2)、(3S,6S)-3-(2-丁基)-6-羟甲基-2,5-哌嗪二酮(3)、(4S)-4-苄基-1,3-恶唑-2,5-二酮(4)、6'-羟基-4,2',3',4'-四甲氧基-对三联苯(5)、(3S,6S)-3-(4-羟苄基)-6-异丁基-2,5-哌嗪二酮(6)、(3R,8aS)-3-异丙基-六氢吡咯并[1,2-a]吡嗪-1,4-二酮(7)、(3S,6S)-3-异丁基-6-甲基-2,5-哌嗪二酮(8)、(3S,6S)-3-苄基-6-(1-羟乙基)-2,5-哌嗪二酮(9)和(3S,6S)-3-苄基-6-(1-羟丙基)-2,5-哌嗪二酮(10)。化合物4目前只见化学合成报道,为新天然产物。     

拟诺卡氏放线菌YIM90087的代谢产物研究

对拟诺卡氏菌YIM90087的固体发酵提取物进行了化学成分研究,从中分离得到10个化合物。通过波谱数据分析,鉴定其结构分别为:(3S)-3-苄基-2,5-哌嗪二酮(1)、(3S,6S)-3-(2-丁基)-6-(1-羟乙基)-2,5-哌嗪二酮(2)、(3S,6S)-3-(2-丁基)-6-羟甲基-2,5-哌嗪二酮(3)、(4S)-4-苄基-1,3-恶唑-2,5-二酮(4)、6'-羟基-4,2',3',4'-四甲氧基-对三联苯(5)、(3S,6S)-3-(4-羟苄基)-6-异丁基-2,5-哌嗪二酮(6)、(3R,8aS)-3-异丙基-六氢吡咯并[1,2-a]吡嗪-1,4-二酮(7)、(3S,6S)-3-异丁基-6-甲基-2,5-哌嗪二酮(8)、(3S,6S)-3-苄基-6-(1-羟乙基)-2,5-哌嗪二酮(9)和(3S,6S)-3-苄基-6-(1-羟丙基)-2,5-哌嗪二酮(10)。化合物4目前只见化学合成报道,为新天然产物。     

白及非多糖组分止血活性成分研究

初步研究白及非多糖组分(BS-80EE)中止血活性成分。采用超高压液相色谱-电喷雾-四级杆-飞行时间串联质谱(UHPLC-ESI-Q-TOF MS)联用技术分析BS-80EE中化学成分;利用二磷酸腺苷(ADP)、凝血酶(THR)诱导的血小板聚集实验和小鼠凝血时间(CT)实验初步评价其止血活性。结果显示,从BS-80EE中共成功指认了12个化合物。其中4-hydroxymethylphenylβ-D-glucoside(1)、blestroside(5)、gymnoside I(7)、gymnosideⅨ(14)、armatuside(17)能显著促进血小板聚集作用。4-Hydroxymethylphenylβ-D-glucoside(1)、α-isobutylmalic acid(3)、blestroside(5)、gymnoside I(7)、militarine(8)、gymnosideⅨ(14)、1-p-hydroxybenzyl-4-methoxyphenanthrene-2,7-diol(18)可显著缩短肝素化小鼠的CT值。该研究结果初步明确了BS-80EE发挥止血的物质基础。     

Liquid chromatography-tandem mass spectrometry analysis of protocatechuic aldehyde and its phase I and II metabolites in rat

A method using liquid chromatography-electrospray ionization tandem mass spectrometry (LC-MS/MS) analysis was established for the identification of metabolites in rat after oral administration of protocatechuic aldehyde, a major bioactive phenolic acid in the roots of Salvia miltiorrhiza. Eleven metabolites in rat plasma and urine were firstly identified as protocatechuic aldehyde, protocatechuic acid and their methylated, glucuronized or glycine conjugates on the basis of their MS fragmentation behaviors, while nine of these metabolites (except protocatechuic aldehyde and protocatechuic acid) were detected in rat bile. In addition, the possible metabolic pathway was proposed for the first time. In the phase I metabolism, protocatechuic aldehyde could be oxidized to protocatechuic acid. The conjugates would be formed in rat intestine, liver and kidney and excreted from rat urine and bile. Enthrohepatic circulation played an important role in the metabolism of protocatechuic aldehyde. The results proved that the established method was simple, reliable and sensitive, revealing that it could be used to rapid screen and identify the structures of active components responsible for pharmacological effects of protocatechuic aldehyde and to better understand its in vivo metabolism.     

降胆固醇乳酸菌对高脂日粮大鼠肠道代谢产物及胆汁酸代谢的研究

通过对肥胖大鼠肠道代谢物改变的研究,揭示乳酸菌降低肥胖大鼠血清胆固醇的机制及对各代谢途径产生的影响。选取健康雄性5周龄SD大鼠并分为3组:饲喂低脂日粮的对照组(C),饲喂高脂日粮的高脂组(H),饲喂高脂日粮同时灌胃乳酸菌BX-1的乳酸菌组(BX-1)。通过气相色谱-质谱联用(GC-MS)非靶向的方式测定各处理组大鼠粪便代谢产物的变化。灌胃乳酸菌BX-1可影响肥胖大鼠肠道代谢物,尤其可促进肥胖大鼠体内泛酸及多种糖代谢中间产物(醛糖、木糖、乳糖、核糖、鼠李糖、果糖)含量的增加。同时影响氨基酸代谢产物脯氨酸的大量增加,伴随焦谷氨酸和鸟氨酸的降低。BX-1还能促进高脂日粮大鼠肠道内不饱和脂肪酸十八烯酸的增加。BX-1对胆汁酸代谢物的影响较大,促进肥胖大鼠肠道内游离胆汁酸以及牛磺类结合胆汁酸的大量生成。胆固醇作为胆汁酸合成的前体物质,有助于血清胆固醇的降低。BX-1主要通过调节肥胖大鼠体内糖代谢、氨基酸代谢以及胆汁酸代谢来降低血清胆固醇含量。     

Influence of glucocorticoid on the metabolism of aldosterone in the isolated perfused rat liver and kidney.

The effect of glucocorticoid deficiency and excess on the extraadrenal metabolism of D-[4-14C]aldosterone (at 4 nM) was studied by radioimmunoassay and by high-performance liquid chromatography in the isolated perfused liver and kidney of adult Wistar rats. Bilateral adrenalectomy was performed 3 weeks before experiments. In nonadrenalectomized rats, 0.3 mg/kg/day dexamethasone was continuously infused subcutaneously for 1 week before experiments. Adrenalectomy did not affect hepatic or renal metabolism of aldosterone. Dexamethasone treatment did not change the renal handling of aldosterone. However, the hepatic clearance of aldosterone was 19% lower (P less than 0.05) in livers of dexamethasone treated rats than in livers of normal rats. After 5 minutes, perfusate [4-14C]aldosterone metabolites were lower in livers of dexamethasone-treated than in livers of normal rats (P less than 0.05). Similar perfusate levels were then obtained. Radiometabolite peaks with similar relative retention times were found in the hepatic perfusate of all groups. However, the ratio between circulating polar metabolites of aldosterone and the metabolites less polar than tetrahydroaldosterone, after 5 and 15 minutes, was highest in livers of dexamethasone-treated rats. Biliary elimination of 14C was similar in all groups. Significant amounts of conjugated tetrahydroaldosterone were only excreted in the bile of dexamethasone-treated rats. In conclusion, glucocorticoid excess reduced the hepatic clearance of aldosterone and changed the pattern of the hepatic metabolites of aldosterone both in circulation and in bile.     

The role of bile and digestive juices in cadmium effect on the absorption processes in rat small intestine

The effects of bile and digestive juices was studied on the intestinal absorption of water, sodium and glucose in the small intestine of rats after their intoxication with one dose of cadmium 1.33 mg/kg of body weight injected intravenously. The investigations were carried out on 60 rats by the method of intestinal perfusion. The obtained results showed that cadmium inhibited the intestinal absorption of these substances. Bile and digestive juices abolished partially this effect during their physiological secretion. After administration of cholagogues no protective role of bile and digestive juices was observed alleviating the toxic effects of cadmium, and the intestinal absorption was even more reduced.     

The mechanisms of intestinal absorption of the carcinogen MNNG (N-methyl-N'-nitro-N-nitrosoguanidine)

We studied the characteristics and mechanisms of MNNG (N-methyl-N'-nitro-N-nitrosoguanidine) intestinal absorption and the interaction between bile acids and fatty acids and MNNG absorption rate in vivo in male Sprague-Dawley rats. We perfused a segment of the proximal small bowel with a physiological solution containing MNNG to assess its basic kinetics and the influence of some physiological and dietary factors on carcinogen absorption. We found that MNNG was absorbed by simple passive diffusion. Transport of MNNG was the highest at pH 6.0. The addition of the bile salt, taurocholate by itself, greatly increased MNNG absorption, while the addition of the long-chain unsaturated fatty acids, oleic and linoleic, decreased the rate of absorption of MNNG. The phospholipid lecithin addition to the perfusate did not change the rate of MNNG absorption. Induction of dietary vitamin A deficiency (serum vitamin A level decreased from 40.9 to 13.7 micrograms/dl) did not change the absorption rate of MNNG. These studies demonstrate that bile acids, dietary fatty acids, and the pH of the intestinal content can modify the rate of absorption of this carcinogen by the small intestine. Since initial intestinal absorption determines serum levels and subsequent reabsorption and enterohepatic cycling determines long-term lumenal levels, serum levels, and total body content, factors which modify the rate of intestinal absorption of MNNG could also modify its carcinogenicity.     

Simultaneous determination of sulforaphane and its major metabolites from biological matrices with liquid chromatography-tandem mass spectroscopy

A simple, sensitive and specific LC-MS/MS method for the simultaneous determination of sulforaphane (SFN) and its major metabolites, the glutathione (SFN-GSH) and N-acetyl cysteine conjugates (SFN-NAC) from biological matrices was developed and validated. The assay procedure involved solid-phase extratcion of all three analytes from rat intestinal perfusate using C2 extraction cartridges, whereas from rat plasma, metabolites were extracted by solid-phase extraction and SFN was extracted by liquid-liquid extraction with ethyl acetate. Chromatographic separation of SFN, SFN-GSH and SFN-NAC was achieved on a C8 reverse phase column with a mobile phase gradient (Mobile Phase A: 10mM ammonium acetate buffer, pH: 4.5 and Mobile Phase B: acetonitrile with 0.1% formic acid) at a flow rate of 0.3 mL/min. The Finnigan LCQ LC-MS/MS was operated under the selective reaction monitoring mode using the electrospray ionization technique in positive mode. The nominal retention times for SFN-GSH, SFN-NAC and SFN were 8.4, 11.0, and 28.2 min,, respectively. The method was linear for SFN and its metabolites with correlation coefficients >0.998 for all analytes. The limit of quantification was 0.01-0.1 microm depending on analyte and matrix, whereas the mean recoveries from spiked plasma and perfusate samples were approximately 90%. The method was further validated according to U.S. Food and Drug Administration guidance in terms of accuracy and precision. Stability of compounds was established in a battery of stability studies, i.e., bench top, auto-sampler and long-term storage stability as well as freeze/thaw cycles. The utility of the assay was confirmed by the analysis of intestinal perfusate and plasma samples from single-pass intestinal perfusion studies with mesenteric vein cannulation in rats.     

In vitro intestinal microflora-mediated metabolism of biliary metabolites from 1-nitropyrene-treated rats

To investigate the modifying role of the intestinal microflora in the metabolism of 1-nitropyrene (1-NP) via enterohepatic circulation, we collected bile from male Wistar rats administered [3H]1-NP orally. The bile was mixed with the intestinal contents (IC) prepared from untreated rats and the mixture was incubated anaerobically under an atmosphere of nitrogen at 37 C. Samples of the reaction mixture were removed at intervals to assay their mutagenic potential, to determine the radioactivity bound to the IC, and for analysis of the biliary metabolites. The binding of the radioactivity to the IC increased linearly as a function of time during the 1-hr incubation. The time-dependent binding does not occur with heat-treated IC and the binding was inhibited by addition of D-saccharic acid 1,4-lacton, a beta-glucuronidase inhibitor. The mutagenicity (for Salmonella typhimurium strain TA98 without S9 mix) of the bile increased early in the incubation period and then decreased very rapidly. The mutagenicity of the bile was also enhanced by treatment with a sonicated IC extract or beta-glucuronidase, but not with a heat-treated IC or aryl-sulfatase. The metabolites produced after the bile was incubated for short periods with the IC were mainly nitrohydroxypyrenes; at later times nitroreduction occurred. The level of acetylaminohydroxypyrenes, which were formed by deconjugation, did not change during the incubation. To determine the degree of contribution of the IC to the total acetylating capacity, we measured acetyltransferase activity of the IC and various organs in Wistar rats. The liver had the highest N-acetyltransferase activity among the seventeen organs examined. Considerable activity was also detected in the kidney, small intestine, lung, and testis, but the IC showed very low activity. The acetylating capacity of the IC was 0.27% of the total capacity in rats, and that of the liver was more than 80%. These results suggest that the nitrohydroxypyrenes formed from 1-NP in the liver were conjugated to glucuronic acid and excreted via the bile duct into intestine. Hydrolysis of these glucuronide conjugates by bacterial beta-glucuronidase liberated into intestine, free nitrohydroxypyrenes, which were direct-acting mutagens. The released aglycons were then rapidly nitro-reduced by intestinal microflora, but contribution of the intestinal microflora to acetylation of the reduced metabolites is very low.     

Structural characterization of metabolites of salvianolic acid B from Salvia miltiorrhiza in normal and antibiotic-treated rats by liquid chromatography-mass spectrometry

This study was conducted to compare the in vivo metabolites of salvianolic acid B (Sal B) between normal rats and antibiotic-treated rats and to clarify the role of intestinal bacteria on the absorption, metabolism and excretion of Sal B. A valid method using LC-MS(n) analysis was established for identification of rat biliary and fecal metabolites. And isolation of normal rat urinary metabolites by repeated column chromatography was applied in this study. Four biliary metabolites and five fecal metabolites in normal rats were identified on the basis of their MS(n) fragmentation patterns. Meanwhile, two normal rat urinary metabolites were firstly identified on the basis of their NMR and MS data. In contrast, no metabolites were detected in antibiotic-treated rat urine and bile, while the prototype of Sal B was found in antibiotic-treated rat feces. The differences of in vivo metabolites between normal rats and antibiotic-treated rats were proposed for the first time. Furthermore, it was indicated that the intestinal bacteria showed an important role on the absorption, metabolism and excretion of Sal B. This investigation provided scientific evidence to infer the active principles responsible for the pharmacological effects of Sal B.