目的 建立超高效液相色谱法(UPLC)测定罗汉果中11-O-罗汉果苷Ⅴ、罗汉果皂苷Ⅴ、赛门苷Ⅰ、罗汉果苷Ⅳ、罗汉果苷Ⅲ、罗汉果苷Ⅲe、罗汉果苷Ⅱe的含量.方法 采用Waters BEH C18色谱柱(50 mm×2.1 mm,1.7μm),流动相为水-乙腈溶液,梯度洗脱;柱温35℃;流速为0.3 mL?min-1;进样量为1μL;采用紫外检测器进行定量测定,检测波长203 nm;采用质谱检测器进行定性鉴别,检测模式为选择离子监测,监测离子的质荷比为1307.8,1309.8,1147.8,1147.8,985.7,985.7和823.7.结果 11-O-罗汉果苷Ⅴ、罗汉果皂苷Ⅴ、赛门苷Ⅰ、罗汉果苷Ⅳ、罗汉果苷Ⅲ、罗汉果苷Ⅲe、罗汉果苷Ⅱe分别在0.50～20.10,5.03～201.20,0.51～20.24,0.52～20.63,0.51～20.22,0.51～20.56,0.50～19.92μg?mL-1范围内线性关系良好(r≥0.999);平均回收率分别为100.7％,98.4％,100.4％,99.0％,100.4％,99.2％和99.2％,RSD分别为1.9％,1.0％,2.0％,1.9％,1.3％,0.8％和2.1％;质谱鉴别结果与紫外检出情况一致.结论 该方法操作便捷、结果准确,重复性好,可用于同时测定罗汉果中的11-O-罗汉果苷Ⅴ、罗汉果皂苷Ⅴ、赛门苷Ⅰ、罗汉果苷Ⅳ、罗汉果苷Ⅲ、罗汉果苷Ⅲe、罗汉果苷Ⅱe的含量.

目的 建立LC-MS/MS测定复方制剂中二甲双胍、吡格列酮及其活性代谢产物酮基吡格列酮(M-Ⅲ)、羟基吡格列酮(M-Ⅳ)的浓度.方法 吡格列酮样品以乙腈沉淀蛋白,美金刚为内标,采用Ultimate C18(2.1 mm×150 mm,5μm)色谱柱,以乙腈-5 mmol·L 1醋酸铵(55∶45)为流动相,流速为0.3 mL· min-1;二甲双胍样品以乙腈沉淀蛋白,卡马西平为内标,采用Ultimate XB-CN色谱柱(2.1 mm×150 mm,5μm),以乙腈-0.1％甲酸水溶液(内含10 mmol·L-1乙酸铵)(73:27)；流速为0.3mL· min-1,柱温:35℃.结果 吡格列酮、M-Ⅲ、M-Ⅳ线性范围分别为5.08 ～1016.0μg·L-1,2.45 ～490.0 μg·L-1,4.20 ～840.0 μg·L-1,定量下限分别为5.08、2.45、4.20 μg·L-1,线性良好.提取回收率均大于89.67％,日内日间RSD均小于15％.结论 本法测定准确、简便可靠,适用于复方二甲双胍吡格列酮片的人体药动学研究.

: A rapid LC-MS/MS method with good accuracy and sensitivity was developed and validated for the pharmacokinetics study of metoprolol (MP) in beagle dogs. The plasma samples were simply precipitated by methanol and then analyzed by LC-MS/MS. An Ultimate XB-C18 column (150 × 2.1 mm ID, 5 μm) was used for separation, with methanol-water containing 0.2% formic acid (65:35, v/v) as the mobile phase at a flow rate of 0.2 mL/min. Monitoring ions of MP and internal standard (hydroxypioglitazone) were m/z 268.1/115.6 and m/z 373.1/150.2, respectively. The linear range was 3.03–416.35 ng/mL with an average correlation coefficient of 0.9996, and the limit of quantification was 3.03 ng/mL. The intra- and inter-day precision was less than 15%. At low, middle and high concentrations, the recovery, the matrix effect and the accuracy was in the range of 76.06%–95.25%, 93.67%–104.19% and 95.20%–99.96% respectively. The method was applied for the pharmacokinetics study of MP tartrate tablets (50 mg). The AUC0-t, Tmax and Cmax were respectively 919.88 ± 195.67 μg/L·h, 0.96 ± 0.33 h, 349.12 ± 78.04 ng/mL.

Objective To evaluate in vitro release behavior of fexofenadine capsules,establish a LC-MS/MS method for the content determination of fexofenadine and study its pharmacokinetics in Beagle dogs.Methods The release behavior was evaluated by using a cup method in four media;a LC-MS/MS method was used to determine drug concentration after a single dose of administration of tablets or capsules in six dogs;pharmacokinetic parameters were calculated with the DAS2.1.1 program.Results The two preparations showed good release behavior;the AUC0-t of fexofenadine capsules and tablets were(20.00±7.48)and(16.42±9.37) mg·L-1·h,AUC0-∞ were(20.30±8.08)and(16.68±9.44) mg·L-1·h,t1/2 were(3.10±1.60)and(4.54±2.03)h,tmax were(1.58±0.49)and(1.67±0.68) h,and Cmax were(4.24±1.69)and(3.68±1.89) mg·L-1,respectively.Conclusion The capsules and tablets of fexofenadine present similar release behavior in vitro.No obvious difference has been found between the main pharmacokinetic parameters of them,which suggests they would have the same therapeutic efficacy.

Objective:To develop an ultra performance liquid chromatography-tandem mass spectrometry method(LC-MS/MS) for the determination of ambroxol in human plasma and its bioequivalence study.Method:Quantitation was performed on a triple quadruple mass spectrometer employing electrospray ionization.The ion pairs for ambroxol and internal standard were monitored at m/z 379.0/263.9 and 237.0/194.1 in positive ion mode,respectively.Carbamazepine was used as the internal standard,and acetonitrile was used to precipitate plasma proteins.The column was Ultimate XB-CN(2.1 mm×150 mm,5μm).The mobile phase consisted of acetonitrile and 0.2%formic acid(40:60) at a flow rate of 0.3 ml·min~(-1).Result:Linearity was within the range of 0.53-187.50 ng·ml~(-1).The lowest quantitation limit was identifiable and reproducible at 0.53 ng·ml~(-1).The Intra- and inter-day RSD were both less than 15%.The accuracy was 101.41%,105.40%and 106.04%,respectively.The extraction recoveries were 82.51%,93.57%and 89.19%,and the matrix effects were 90.48%,92.36%and 100.81%,respectively.The stability tests showed the promising stability of the samples during the whole study.Conclusion:A rapid,sensitive and simple LC-MS/MS method for determination of ambroxol in human plasma is developed,which is successfully applied in its bioequivalence study.

目的:评价2种匹伐他汀钙片在人体内的生物等效性。方法:20例男性健康志愿者随机交叉单剂量口服试验制剂和参比制剂,利用LC-MS/MS法测定匹伐他汀体内血药浓度。结果:试验制剂和参比制剂的主要药动学参数如下:AUC0-t分别为（180.2±37.6）μg.h.L-1和（182.6±36.1）μg.h.L-1;AUC0-∞分别为（190.0±38.1）μg.h.L-1和（189.6±37.4）μg.h.L-1;Cmax分别为（54.3±18.2）μg.L-1和（65.1±22.4）μg.L-1;tmax分别为（0.80±0.25）h和（0.78±0.26）h;t1/2分别为（13.4±4.5）h和（12.0±2.8）h;经方差分析和双单侧t检验显示,主要药动学参数无显著差异,以AUC0-t计算匹伐他汀钙片的相对生物利用度为（100.3±19.9）%。结论:试验制剂与参比制剂具有生物等效性。

目的 研究泽泻总三萜提取物对大鼠尿草酸钙结石形成的影响,从而确认泽泻抗泌尿系结石的活性成分.方法 在实验的28 d内给予乙二醇及阿法骨化醇诱导雄性SD大鼠泌尿系草酸钙结石形成.造模的同时分别给予不同剂量的泽泻总三萜提取物,分别为205.2、68.4及22.8 mg/kg.实验结束时,检测各组大鼠体重,血尿素氮(BUN)、肌酐(Cr)、P、Ca2+、Mg2+含量,24 h尿量,尿pH值,24 h尿Ca2+、Mg2+、尿草酸(Ox)的分泌量,右肾系数,肾Ca2+、Mg2+含量;显微镜下观察肾组织切片中草酸钙结晶沉积及病理变化,评价泽泻总三萜对泌尿系结石形成的影响.结果 乙二醇及阿法骨化醇诱导的泌尿系草酸钙结石大鼠血BUN、Cr、24 h尿Ox、尿Ca2+分泌量、肾Ca2+含量及右肾系数明显升高,肾Mg2+含量明显下降.泽泻总三萜提取物能降低泌尿系结石大鼠的血BUN、Cr、24 h尿Ca2+含量、肾Ca2+含量及右肾系数水平,增加24 h尿量及肾组织Mg2+含量.肾组织病理学检查可知泽泻总三萜能减少大鼠肾组织内草酸钙晶体的沉积,改善肾脏组织的损伤情况.结论 泽泻总三萜提取物可以有效抑制乙二醇及阿法骨化醇诱导的大鼠泌尿系草酸钙结石的形成,泽泻总三萜是泽泻抗泌尿系结石的活性成分.

Objective To study the effects of the total triterpenoids extract of Alismatis Rhizoma on calcium oxalate urinary stone formation in rats,thereby confirming the anti-urolithiatic active constituents of Alismatis Rhizoma.Methods Adult male Sprague Dawley rats were given ethylene glycol and alfacalcidol to induce urolithiasis for 28 days.Meanwhile,the rats received the total triterpenoids extract at the doses of 205.2,68.4,22.8 mg/kg body weight,respectively.On the 28th day,body weight,the levels of serum urea,creatinine,P,Ca2+ and Mg2+,24 h urine volume,urine pH,Ca2+,Mg2+ and oxalate,relative kidney weight and renal Ca2+,Mg2+ were evaluated to estimate the anti-urolithiatic effects of the total triterpenoids extract.Kidney sections were examined under a light microscope to study the calcium oxalate crystal deposition and pathological changes.Results The urolithiasis induced by ethylene glycol and alfacalcidol not only caused significant increase in the levels of serum urea and creatinine,urine oxalate and Ca2+,relative kidney weight and renal Ca2+,but also the reduction in renal Mg2+.The administration of the total triterpenoids extract reversed the changes in the levels of serum urea and creatinine,urine Ca2+,relative kidney weight,renal Ca2+ and Mg2+.It also diminished the number and size of calcium oxalate crystal deposits and protected the kidneys from serious damage.Conclusion The total triterpenoids extract of Alismatis Rhizoma has a beneficial effect against calcium oxalate urinary stone formation induced by ethylene glycol and alfacalcidol.The total triterpenoids are the anti-urolithiatic active constituents of Alismatis Rhizoma.

OBJECTIVE To evaluate the bioequivalence of two preparations of diacerhein in healthy volunteers. METHODS 20 healthy volunteers were randomly divided into two groups and taken a single oral dose of diacerhein of test and reference preparation respectively. The concentration in plasma was determined by LC-MS/MS. RESULTS The main pharmacokinetic parameters of test and reference were as follows: AUC_(0-t) (25 205. 6 ± 15 694. 9)μg·h·L~(-1) and (24 927. 7 ± 15 869. 8)μg·h· L~(-1) ;AUC_(0-∞)(26 266.3±17 021.4)μg·h·L~(-1)and (259 61. 7 ± 16 592. 5)μg·h·L~(-1);C_(max) (4 276. 8± 2 391. 8)μg·L~(-1) and (4 352. 8 + 2 415. 4)μg·L~(-1);t_(max)(2. 3± 0. 7)h and(2. 3 ± 0. 8)h;t_(1/2) (4. 5 ± 1. 4)h and(4. 4 ± 0. 9)h;The relative bioavailability of diacerhein was (105. 2 ± 22. 6) % 。CONCLUSION The test and reference preperation of diacerhein are bioequivalent.

A sensitive, rapid and simple liquid chromatography-electrospray ionization mass spectrometry (LC-ESI-MS/MS) method was developed for the quantitative determination of cyclobenzaprine in human plasma, to study the pharmacokinetic behavior of cyclobenzaprine capsule in healthy Chinese volunteers. With escitalopram as the internal standard (IS), sample pretreatment involved a one-step liquid-liquid extraction using saturated sodium carbonate solution and hexane-diethyl ether (3:1, v/v). The separation was performed on an Ultimate XB-CN column (150 × 2.1 mm, 5 µm). Isocratic elution was applied using acetonitrile-water (40:60, v/v) containing 10 m M ammonium acetate and 0.1% formic acid. The detection was carried out on a triple-quadrupole tandem mass spectrometer in multiple reaction monitoring mode via electrospray ionization. The ion-pairs including m/z 276.2-216.2 for cyclobenzaprine and m/z 325.2-109.1 for IS were used for monitoring. Linear calibration curves were obtained over the range of 0.049-29.81 ng/mL with the lower limit of quantification at 0.049 ng/mL. The intra- and inter-day precision showed ≤ 6.5% relative standard deviation. The established method laid the groundwork for follow-up studies and provided basis for the clinical administration of cyclobenzaprine.

A sensitive, simple and rapid ultra fast liquid chromatography (UFLC)-ESI-MS/MS method was established for the simultaneous determination of 3,3',4',5,7-pentamethylquercetin (PMQ) and its possible metabolite 3,3',4',7-tetramethylquercetin (TMQ) in dog plasma using 4',5,7-trimethylapigenin (TMA) as the internal standard. The plasma sample was pretreated with acetonitrile for protein precipitation and the analytes were separated on an Ultimate XB-CN column (5 μm, 2.1 mm × 150 mm) with the mobile phase consisting of acetonitrile and water (2:1, v/v). Detection was performed on a triple-quadrupole tandem mass spectrometer under a positive multiple reaction-monitoring mode (MRM). The mass transition ion-pair was followed as m/z 373.1-312.1 for PMQ, 359.1-344.0 for TMQ and 313.1-298.1 for TMA. The validated concentration ranged from 1.272 to 3060 ng/mL for PMQ and from 10.35 to 1725 ng/mL for TMQ. The lower limit of quantifications for PMQ and TMQ were 1.272 ng/mL and 10.35 ng/mL, respectively. The developed-method was successfully applied for the pharmacokinetic study of PMQ and its metabolite TMQ in dogs following a single oral dose.

OBJECTIVE To study the pharmacokinetics and bioequivalence of two preparations of nicergoline tablet in healthy volunteers.METHODS In a two-way cross-over study,single oral dose of nicergoline tablets of test and reference preparation was given to 20 healthy volunteers respectively.Nicergoline concentration in plasma was determined by HPLC-MS/MS.RESULTS The main pharmacokinetic parameters of nicergoline major metabolite 10α-methoxy-9,10-dihydrolysergol(MDL) of nicergoline tablets(test and reference) were as follows:AUC(0-t)(502.559±119.468) and(487.320±118.702)μg.h.L-1;AUC(0-∞)(515.650±124.198) and(502.537±126.780)μg.h.L-1;Cmax(32.445±8.203)and(30.590±6.760)μg.L-1;Tmax(4.300±1.780)and(3.650±1.565)h;T1/2(10.969±1.641)and(11.473±2.054)h.The F(0-t)and F(0-∞) of the test tablets were(104.9±23.5)% and(104.4±23.0)%.CONCLUSION The test and reference preparation of nicergoline tablets are bioequivalent.

OBJECTIVE To establish an UFLC-MS/MS method for determination ot tiopronin in human plasma and investigating the pharmacokinetics and bioequivalence of tablets and enteric-coated capsules in healthy volunteers.METHODS Twenty volunteers were randomly divided into two groups(test and reference)in a double cross-over design.Pharmacokinetic parameters were calculated with DAS 2.0 program.RESULTS The linear range was 4.05-8 100 μg·L-1,and the limit of determination was 4.05 μg·L-1.The method recoveries were 85.0%-91.3% and the RSD of within-day and between-day was less than 15%.The main pharmacokinetic parameters of tiopronin tablets and enterosoluble capsules were as follows:Cmax(2 151.1±1 135.0) μg·L-1 and (2 363.0±1 055.1)μg·L-1;tmax(4.8±1.3) h and (4.3±1.2)h;AUC(0-t)(11 618.2±3 625.5) μg·h·L-1 and (12 824.0 ±4 464.1) μg·h·L-1;AUC(0-∞) (12 677.7±3 874.9) μg·h·L-1 and (13 803.1±4 686.0) μg·h·L-1.CONCLUSION The method is simple,sensitive and specific and the statistical analysis showed that the test and reference preparation were bioequivalent.

目的建立测定普通针毛蕨地下部分中Protoapigenone的梯度洗脱反相高效液相色谱法。方法采用EurospherC18色谱柱(250mm×4.0mm,5μm),甲醇-水(0.1%磷酸,pH值=3.0)为流动相,梯度洗脱,检测波长250nm,柱温25℃。结果Protoapigenone在7.5～480.0mg·mL-1范围里线性关系良好,普通针毛蕨地下部分中Protoapigenone含量为1.14%,RSD为3.5%。结论该法稳定、简便,可用于普通针毛蕨药材的质量控制。