Transport and uptake characteristics of a new derivative of berberine(CPU-86017) by human intestinal epithelial cell line: Caco-2

AIM: The characteristics of transepithelial transport and uptake of CPU-86017 {[7-(4-chlorbenzyl)-7,8,13,13α-tetrahydroberberine chloride, CTHB]}, a new antiarrhythmia agent and a new derivative of berberine, were investigated on epithelial cell line (Caco-2) to further understand the absorption mechanism of berberine and its derivatives. METHODS: Caco-2 cell was used. RESULTS: 1) The permeability coefficient from the apical (AP) to basolateral (BL) of CPU-86017 was approximately 5 times higher than that from BL-to-AP transport. The effects of a P-glycoprotein (P-gp) inhibitor-cyclosporin A, some surfactants, and lower pH on the transepithelial transport of CPU-86017 were also observed. Cyclosporine A at 7.5 mg/L had no effect on the transepithelial electrical resistance (TEER); an about 4-fold enhancement on the transepithlial transport of CPU-86017 was observed. Some surfactants (sodium citrate, sodium deoxycholate, and sodium dodecyl sulfate) at 100 μmol/L and low pH (pH=6.0) induced a reversible d     

4′-甲基-7-(2-羟基-3-异丙胺基-丙氧基)-黄酮盐酸盐对冠脉血流和家兔实验性心肌梗塞的影响

SIPI-549 is a new cOmpound. In Langendorff's isolated rabbit heart, SIPI-549 significantly increased coronary blood flow (p<0.01), but did not change themyocardium contractibility and heart rate. A study of the uptake of ~(86)Rb by the mouse heart showed a significant increase of nutritional blood flow in the SIPI-549 treated group (p<0.05). On intact dogs, SIPI-549 also showed a significant increase of coronary blood flow (p<.05) and decrease of coronary resistance (p<0.01), but no change of cardiac output and blood presure was observed.In the model of myocardial infarction by high level double-ligation of the anterior descending branch of the left coronary in rabbits, iv SIPI-549 (4.1 mg/kg) significantly reduced the ΣST, NST and NO The size of infarct myocardium was significantly reduced (p<0.05) 24 h after ligation of the coronary artery Similar results were found in the propranolol (1 mg/kg) group. However, no obvious difference between the two groups was observed.     

Species differences in the CYP3A-catalyzed metabolism of TPN729, a novel PDE5 inhibitor

TPN729 is a novel phosphodiesterase 5(PDE5)inhibitor used to treat erectile dysfunction in men.Our previous study shows that the plasma exposure of metabolite M3(N-dealkylation of TPN729)in humans is much higher than that of TPN729.In this study,we compared its metabolism and pharmacokinetics in different species and explored the contribution of its main metabolite M3 to pharmacological effect.We conducted a combinatory approach of ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry-based metabolite identification,and examined pharmacokinetic profiles in monkeys,dogs,and rats following TPN729 administration.A remarkable species difference was observed in the relative abundance of major metabolite M3:i.e.,the plasma exposure of M3 was 7.6-fold higher than that of TPN729 in humans,and 3.5-,1.2-,1.1-fold in monkeys,dogs,and rats,respectively.We incubated liver S9 and liver microsomes with TPN729 and CYP3A inhibitors,and demonstrated that CYP3A was responsible for TPN729 metabolism and M3 formation in humans.The inhibitory activity of M3 on PDE5 was 0.78-fold that of TPN729(The IC50 values of TPN729 and M3 for PDE5A were 6.17±0.48 and 7.94±0.07 nM,respectively.).The plasma protein binding rates of TPN729 and M3 in humans were 92.7%and 98.7%,respectively.It was astonishing that the catalyzing capability of CYP3A4 in M3 formation exhibited seven-fold disparity between different species.M3 was an active metabolite,and its pharmacological contribution was equal to that of TPN729 in humans.These findings provide new insights into the limitation and selection of animal model for predicting the clinical pharmacokinetics of drug candidates metabolized by CYP3A4.     

Effect of Clkl gene on level of Aβ-induced autophagy in Alzheimer's disease北大核心CSCD

Aim To investigate the effect of siRNA transfection of silencing Clkl gene on autophagy levels in AD model cells. Methods The Clkl gene was silted using siRNA transfection techniques. MTT was used to observe the effects of Aβ2 5 35 and Clkl genes on cell survival. Cellular immunofluorescence was applied to observe the expression of the autophagy marker microtubule-associated protein 1 light chain 3 (LC3). Western blot was employed to detect the expression of Clkl, HIF-1α, PI3K/AKT/mTOR, autophagy-related proteins Beclinl,LC3-II/I,and p62. Quantitative realtime polymerase chain reaction (RT-qPCR) was utilized to detect the expression of Beclinl, LC3, p62 mRNA. Results Compared with the NC + NS group, the viability of HT22 cells in the NC + Aβ group was significantly reduced (P < 0.01), while the viability of cells in the siClkl + Aβ group was significantly higher than that in the N C + A β group (P < 0.01). The results of cellular immunofluorescence showed that the expression of LC3 in the NC + Aβ group was significantly higher than that in the NC + NS group (P < 0.01), and the expression of LC3 in the siClkl + Aβ group was significantly lower than that in the NC + Aβ group (P < 0.01). Compared with the NC + NS group, the expression of Beclinl and LC3 -II/I proteins in the NC + Aβ group was significantly raised (P < 0.01), and the expression of p62, PI3 K, p-AKT/AKT, and p-mTOR/mTOR was significantly reduced (P < 0.01). Compared with the NC + Aβ group, the expression of Beclinl and LC3-II/I proteins in the siClkl + Aβ group decreased significantly (P <0.01), and the expression of p62, PI3 K, p-AKT/AKT, and p-mTOR/mTOR proteins increased significantly (P < 0.01). The addition of LY294002 and YC-1 reversed the effect of silencing Clkl on Beclinl, LC3-II/I, p62, PI3K, p-AKT/AKT, p-mTOR/mTOR proteins. RT-qPCR results showed that there was no significant difference in Beclinl, LC3 and p62 mRNA levels in the siClkl + Aβ + YC-1 group compared with the siClkl + Aβ group (P > 0.05), and Beclinl, LC3 and p62 mRNA were consistent with the protein levels in the other groups. Conclusions Silencing Clkl expression can alleviate the decrease in cell viability induced by excessive autophagy of HT22 cells induced by Aβ2 535, and the mechanism may be related to the PI3 K/AKT/mTOR signaling pathway and HIF-1α. © 2023 Publication Centre of Anhui Medical University. All rights reserved.     

(-)和(+)棉酚对雄大鼠生育力的影响

Racemic, (-) and (+) gossypol, provided by the Department of Organic Chemistry of our institute, was suspended in 2.5% tween 80 solution. Adult male Wistar rats 190～220 g in weight were allotted to 5 groups. Animals in group 1 received 2.5% tween 80 solution as control. Rats in group 2 were treated with racemic gossypol at the dosage of 30 mg/kg for 2 weeks. Animals in group 3 and 4 were given 15 mg/kg of (-) gossypol for 2 weeks and 30 mg/kg of (-) gossypol for 1 week respectively. Rats in group 5 were treated with (+) gossypol at the dosage of 30 mg/kg for 2 weeks.Four weeks from the beginning of gossypol treatment the rats were cohabited with adult females for 7 days. Then the motility of the sperms in the cauda epididymides was estimated The female rats were examined for pregnancy 7 days later.Treatment with (-) gossypol at 30 mg/kg caused significant decreases in body weight of the rats (P<0.05). One of the five rats died 7 days after the last administration, while (+) gossypol and racemic gossypol at the dosage employed had no effect on the body weight. (+) Gossypol at 30 mg/kg for 2 weeks had no effect on the motility of the sperms in the cauda epididymides and no effect on the fertility of the animals: nor was there any effect on the weights of the testis, epididymis, prostate and seminal vesicle. The sperms of the cauda epididymides were found to be dead in the groups treated with 15 and 30 mg/kg of (-)gossypol. Raccmic gossypol given for 2 weeks at 30 mg/kg caused loss of fertility of the male rats which confirmed our previous findings.(?)t may be postulated that (+) gossypol has no antifertility effect nor toxicity at the dosage employed. (-) Gossypol is the active stereoisomer of racemic gossypol.     

急性结石性胆囊炎急诊手术与延期手术的疗效比较

Objective Through comparing the curative effects of urgent and delayed laparoscopic cholecystectomy (LC) on acute calculous cholecystitis,evaluating the safety and feasibility of urgent LC for acute calculous cholecystitis.Mthods 109 patients with a clinic diagnosis of acute calculous cholecystitis were randomly assigned to urgent LC (urgent group,n=55) or delayed interval surgery after initial medical treatment (delayed group,n=54,but with some reasons there only 40 patients were treated by delayed surgery).According to the operation time within 72 hours of admission or not,the urgent group was divided into two groups:37 patients in Group Ⅰ (within 72 hours) and 18 patients in Group Ⅱ (beyond 72 hours).The operation time,bile duct injury,conversion to open cholecystectomy,total hospital stay,postoperative complications and mortality of the perioperative period were compared.Results There was no difference between the urgent LC group and delay LC group in the operating time[ (91±26.2 )min versus( 104±34.7 )min],bile duct injury (1 case versus 0 case),conversion to open cholecysteetomy (3case versus 4 case),total hospital stay[ (8±2.7) day versus( 8±1.2 )day]and postoperative complications (2 case versus 1 case).The delay in performing an LC beyond 72 h affected the operating time and conversion to open eholeeystectomy.No death in perioperative period.Conclusion Urgent LC for acute calculous cholecystitis can be performed safely and successfully.Earlier urgent LC has a beneficial impact for patients.     

用2H-标记物和GC-MS分离鉴定联苯双酯在大鼠尿中的一个代谢产物

Dimethyl-4, 4′-dimethoxy-5, 6, 5′, 6′-dimethylenedioxybiphenyl-2, 2′-dicarboxylate (biphenyldimethyldicarboxylate; BDD), a synthetic compound, has been used in the treatment of chronic hepatitis with good results in reducing s-GPT. Previous work in our laboratory studied its metabolites using ³H-labeled compound in combination with TLC and found that its main methabolic pathway is demethylation followed by conjugation with glucuronic acid. This paper reports the isolation and identification of a metabolite of BDD from rat urine using ²H-labeled compound and GC-MS. Rats fasted for 12h were intragastrically given a mixture of ²H-labeled (consisting of monodeutero-and dideutero-BDD in the ratio about 1:1.3)and non-labeled BDD 150mg/kg and placed in metabolism cages for urine collection. The 24h urine was filtered and extracted three times each with 5ml of methylenedichloride. The extracts were pooled and evaporated to dryness under reduced pressure at 35℃. The residue was redissolved in chloroform and subjected to GC-MS analysis. The mass spectrum (m/z: 404, 405, 406; 373, 374, 375; 345, 346, 347; 330, 331, 332; etc)indicates that the molecular ionic and fragment peaks of the metabolite all have 14 amu less than those of BDD. This means that the metabolite isolated is mono-O-demethylated BDD. The result confirmed our findings reported previously.     

Biotransformation of metoprolol by the fungus Cunninghamella blakesleeana

Aim： To investigate the biotransformation of metoprolol, a β-cardioselective adrenoceptor antagonist, by filamentous fungus, and to compare the parallels between microbial transformation and mammalian metabolism. Methods： Five strains of Cunninghamella （ C elegans AS 3.156, C elegans AS 3.2028, C echinulata AS 3.2004, C blakesleeana AS 3.153 and AS 3.910） were screened for the ability to transform metoprolol. The metabolites of metoprolol produced by C blakesleeana AS 3.153 were separated and assayed by liquid chromatography-tandem mass spectrometry （LC/MSn）. The major metabolites were isolated by semipreparative HPLC and the structures were identified by a combination of LC/MSn and nuclear magnetic resonance analysis. Results： Metoprolol was transformed to 7 metabolites; 2 were identified as new metabolites and 5 were known metabolites in mammals. Conclusion： The microbial transformation of metoprolol was similar to the metabolism in mammals. The fungi belonging to Cunninghamella species could be used as complementary models for predicting in vivo metabolism and producing quantities of metabolite references for drugs like metoprolol.     

Anti-diarrheal and spasmolytic activities study of 3,4-Dihydro-2-( 4-Morpholinylmethy )-1 (2H)-Naphthalenone(CY)

Objective To investigate the spasmolytic and anti-diarrheal activities of 3,4-dihydro-2-(4-morpholinylmethy)-1(2H)-naphthalenone(CY),which was first synthesized by Welch,Willard M et al.in 1977.Methods The spasmolytic effects of CY were tested on isolated rabbit small intestine at the concentration of 0.01 to 3 mM;the diarrheal-index was evaluated on diarrhea mice to study the anti-diarrheal activities of CY.Results CY(0.1-1 mM)inhibited spontaneous motility of rabbit small intestine and at the concentration of 0.01 to 3 mM CY inhibited the contractile response of rabbit small intestine and colon induced by acetylcholine(10-2 mg·mL-1),high K+(60 mM)and BaCl2(1 mg·mL-1).When tested against calcium channel blocked in rabbit small intestine and colon,CY caused a rightward shift in the Ca2+ dose-response curves,similar to that produced by verapamil,a well-known calcium antagonist.CY could inhibit the diarrhea induced by castor oil,MgSO4 and liquid paraffin and LD50 of CY is 277.2 mg·kg-1.Conclusions CY may produce its spasmolytic and anti-diarrheal effects as a calcium antagonist.     

6-取代二氢哒嗪酮类化合物的合成及血小板聚集抑制作用

In order to develop more potent and less toxic, antithrombotic agents, ten 6-(4-substituted piperazinyl acetyl aminophenyi)-4, 5-dihydro-3(2H)-pyridazinones were synthesized. The title compounds were tested in vitro for platelet aggregation inhibitory activity with ADP-induced rat platelets and PAF-induced rabbit platelets. Preliminary tests showed that all of the pyridazinones could inhibit ADP-induced rat platelet aggregation. Ⅰ₇, Ⅰ₈, Ⅰ₉ were more potent than the control compound CI 930. Ⅰ₉ was the most potent compound with IC₅₀ of 0.99 μmol/L. Pertaining to PAF-induced rabbit platelet aggregation. Ⅰ₉ was the most potent inhibitor with IC₅₀ of 3.7 μmol/L.     

7-(4-氯苄基)-7,8,13,13a-四氢小檗碱在家兔体内的代谢产物分析

7(4氯苄基)7,8,13,13ａ四氢小檗碱(ＣＴＨＢ)结构式为:是中国药科大学对小檗碱进行结构改造得到的新化合物,有较强的抗心律失常作用,已在多个国家申请了专利。我们曾对该化合物进行了Ｃ,Ｈ化学位移的全归属[1],鉴定了在大鼠体内的主要代谢产物[2]。为了比较该化合物在不同动物体内的代谢情况,我们用家兔进行试验,前文[3]已经报道了用ＨＰＬＣ/ＤＡＤ对家兔胆汁中7(4氯苄基)7,8,13,13ａ四氢小檗碱代谢产物的鉴别,本文进一步研究该化合物在家兔体内的代谢途径。材料与方法　　动物　家兔,重25ｋｇ,由本校动物房提供。　　仪器　ＨＰ1100ＨＰＬ…     

7-(4-氯苄基)-7,8,13,13a-四氢小檗碱氯化物在犬体内的药代动力学和药效学

以 QT校正 ( QTc)延长百分率为效应指标 ,用药代动力学 -药效学 ( PK- PD)结合模型对家犬 iv1 .0 ,2 .0 mg· kg-1的 7- ( 4 -氯苄基 ) - 7,8,1 3,1 3a-四氢小檗碱氯化物 ( CPU860 1 7)后在体内的处置和效应作定量分析 .给家犬 iv CPU860 1 7后的血药浓度经时过程符合二房室模型 ,药物的效应与效应室浓度之间的关系符合 S形 Emax模型 .CPU860 1 7的 PK- PD性质在所用剂量范围内均为非剂量依赖性 .     

LC/DAD/MSD技术研究大鼠胆汁中盐酸非洛普的II相代谢产物

目的研究大鼠服药后胆汁中盐酸非洛普(DDPH)II相代谢产物.方法收集大鼠空白胆汁及给药后12h内的胆汁,以LC/DAD/MSD技术判断II相代谢产物峰位.以HPLC法制备II相代谢产物馏分并以β-葡糖醛酸酶水解,再进行分析;同时将与II相代谢产物相应的I相代谢产物对照品按相同条件进行分析对照.结果大鼠给药后胆汁色谱图中峰M₇,M₈和M₉为DDPH的II相代谢产物,它们的β-葡糖醛酸酶水解产物分别为M₃,M₂和M₁.结论β-1-O-{3,5-二甲基-4-[-2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₇),β-1-O-{2,4-二甲基-3-[2-甲基-2-(3,4-二甲氧基苯乙氨基)-乙氧基]-苯基}-葡糖醛酸(M₈)和β-1-O-{2-甲氧基-4-[1-甲基-2-(2,6-二甲基苯氧基)-乙氨基-乙基]-苯基}-葡糖醛酸(M₉)为大鼠ipDDPH后产生的II相代谢产物.     

LC/DAD/MSD技术研究大鼠服药胆汁中盐酸非洛普I相代谢产物

目的　研究大鼠服药后胆汁中盐酸非洛普(DDPH)I相代谢物。方法　大鼠做胆管插管,分别收集ipDDPH之前的空白胆汁及服药后12h内的服药胆汁,将大鼠胆汁以葡糖醛酸酶水解后进C-18SPE小柱进行纯化富集,再进行LC/DAD/MSD分析;同时将合成的6个DDPH模拟代谢物M₁-M₆的对照品混合液按相同条件进行LC/DAD/MSD分析对照。结果　大鼠服药胆汁色谱图中峰A,B,C,D,E和F分别与M₁,M₂,M₃,M₅,M₄和M₆的保留时间、紫外吸收光谱、分子量及碎片离子完全一致。结论　M₁,M₂,M₃,M₄,M₅和M₆为大鼠ipDDPH后产生的体内I相代谢物。     

LC/MS分析大鼠体内氧化苦参碱及其主要代谢物LC/MS分析大鼠体内氧化苦参碱及其主要代谢物

目的研究氧化苦参碱在大鼠体内的主要代谢产物。方法以氧化苦参碱和苦参碱为对象优化液相色谱/电喷雾离子阱质谱(LC/ESI-ITMSⁿ)实验条件，分析总结其电喷雾质谱的一级电离规律和二级质谱裂解规律，作为氧化苦参碱大鼠体内代谢物结构分析的依据。健康大鼠腹腔肌注40 mg·kg^-1氧化苦参碱，收集0~24 h的尿样，尿样中的代谢物经C₁₈小柱进行富集与纯化后，在优化的LC/ESI-ITMSⁿ条件下进样分析。代谢物的结构推导主要依据代谢物的色谱保留时间及其电喷雾离子阱质谱(ESI-ITMSⁿ)电离规律。结果在大鼠尿样中有原药及其6种I相氧化及还原代谢产物，且主要代谢物为苦参碱。未发现II相代谢物。结论本法不仅操作简便、快速，而且灵敏度高、专属性强。该分析技术是研究药物代谢最有效的方法之一。     

Characterization of a new rat urinary metabolite of piperine by LC/NMR/MS studies

Potential of piperine, an active alkaloid of black and long peppers, to increase the bioavailability of drugs in humans is of great clinical significance owing to its omnipresence in food. In an attempt to further study the reported differences in its metabolism in rats and humans, a new major urinary metabolite was detected in rat urine and plasma using HPLC. The metabolite was partially purified using reverse phase column chromatography on Sephadex^®-LH 20 and characterized as 5-(3, 4-methylenedioxy phenyl)-2E,4E-pentadienoic acid-N-(3-yl propionic acid)-amide with the help of LC/NMR/positive ESI–MS studies. Complete mass fragmentation pattern could be assigned with MS/MS studies. The metabolite has a unique structure compared to the previously reported metabolites in that it retains methylenedioxy ring and conjugated double bonds while the piperidine ring is modified to form propionic acid group. Mechanism of formation of the metabolite by oxidation and cleavage of piperidine ring is proposed. Kidney appears to be the major excretion route for piperine metabolites in rats as no metabolite could be detected in feces.     

Development of LC/MS/MS assay for the determination of 5-ethyl-2-{5-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]-2-propoxyphenyl}-7-propyl-3,5-dihydropyrrolo[3,2-d]pyrimidin-4-one (SK3530) in human plasma: application to a clinical pharmacokinetic study

5-Ethyl-2-{5-[4-(2-hydroxyethyl)piperazine-1-sulfonyl]-2-propoxyphenyl}-7-propyl-3,5-dihydropyrrolo[3,2-d]pyrimidin-4-one (SK3530) is a new phosphodiesterase type-5 inhibitor currently undergoing a Phase III investigation for the treatment of male erectile dysfunction. This study first describes a rapid and sensitive LC/MS/MS assay method for the quantification of SK3530 and its major metabolite, SK3541, in human plasma. The assay was validated to demonstrate the specificity, linearity, recovery, lower limit of quantification (LLOQ), accuracy, and precision. The multiple reaction monitoring was based on the transition of m/z=532.5-->99.1 for SK3530, 488.6-->295.5 for SK3541, and 520.3-->99.1 for SK3304 (internal standard). The assay utilized a single liquid-liquid extraction and isocratic elution, and the LLOQ was 1 ng/ml using 0.2 ml human plasma. The assay was linear over a range from 1 to 1000 ng/ml for both SK3530 and SK3541, with correlation coefficients >0.9999. The mean intra- and inter-day assay accuracy ranged from 94.7 to 101.6% and 96.8 to 101.1% for SK3530 and 92.6-105.7% and 97.4-107.8% for SK3541, respectively. The mean intra- and inter-day precision was between 7.2-12.1% and 5.7-7.4% for SK3530 and 4.6-13.2% and 5.0-14.1% for SK3541, respectively. The developed assay was applied to a clinical pharmacokinetic study after oral administration of SK3530 in healthy male volunteers (dose 100 mg).     

左旋黄皮酰胺在大鼠体内的排泄

左旋黄皮酰胺[（-）-clausenamide]是从芸香科黄皮属植物黄皮[Clausena lansium（lour） sheels]叶的水浸膏中分离得到的有效成分，经不对称合成和拆分制备而得。药效学研究表明，左旋黄皮酰胺促进突触体谷氨酸释放，增加大鼠脑皮层厚度和海马CAL区突触数及NMDA受体密度，提高小鼠脑皮层和海马的胆碱乙酰转移酶活性，对抗樟柳碱引起的乙酰胆碱含量降低。这些结果表明左旋黄皮酰胺具有较好的促智和神经保护作用以及潜在的抗老年痴呆作用；右旋体作用不明显，且有较强的毒性。     

一种新的小檗碱衍生物CPU-86017在人小肠上歧细胞（Caco-2细胞）中的转运与摄取特性

AIM: The characteristics of transepithelial transport and uptake of CPU-86017 [[7-(4-chlorbenzyl)-7,8,13,13alpha-tetrahydroberberine chloride, CTHB]], a new antiarrhythmia agent and a new derivative of berberine, were investigated on epithelial cell line (Caco-2) to further understand the absorption mechanism of berberine and its derivatives. METHODS: Caco-2 cell was used. RESULTS: 1) The permeability coefficient from the apical (AP) to basolateral (BL) of CPU-86017 was approximately 5 times higher than that from BL-to-AP transport. The effects of a P-glycoprotein (P-gp) inhibitor-cyclosporin A, some surfactants, and lower pH on the transepithelial transport of CPU-86017 were also observed. Cyclosporine A at 7.5 mg/L had no effect on the transepithelial electrical resistance (TEER); an about 4-fold enhancement on the transepithlial transport of CPU-86017 was observed. Some surfactants (sodium citrate, sodium deoxycholate, and sodium dodecyl sulfate) at 100 micromol/L and low pH (pH=6.0) induced a reversible decrease of TEER; enhancements of the transepithelial transport of CPU-86017 were also observed with some surfactants; 2) In the process of uptake of CPU-86017, the initial uptake rates of CPU-86017 were saturable with a Vmax of (250+/-39) microg/min/g (protein) and Km of (0.90+/-0.12) mmol/L. This process was enhanced by cyclosporine A (7.5 mg/L) with a Vmax of (588+/-49) microg/min/g (protein) and Km (0.42+/-0.08) mmol/L. CONCLUSION: Some surfactants and P-gp inhibitors can be considered as enhancers of its transepithelial transport and uptake.     

5－乙基－5－对氟苯甲酰丙基巴比妥酸在兔体内的代谢产物研究

用高效液相色谱法对５－乙基－５－对氟苯甲酰丙基巴比妥酸在家兔体内的代谢物进行了分离，用二极管阵列检测器测定了原药及代谢物紫外图谱，得出代谢物的紫外吸收性质；同时测试了原药及代谢物的质谱，推测出代谢物的可能结构，并用化学方法合成了代谢物对照品，通过核磁和质谱的测定进一步确证了结构。