反相离子对高效液相色谱法测定SD大鼠血浆中河豚毒素的含量

建立了大鼠血浆中河豚毒素(TTX)反相离子对高效液相色谱测定方法。血浆样品加水涡漩,再加沉淀剂V(乙腈)∶V(含0.5%HAc的甲醇液)=3∶1),旋涡离心后取上清液进样测定。色谱柱为Agilent ZorbaxSB C18柱(150 mm×4.6 mm×5μm);流动相为10?N-90%的8 mmol/L庚烷磺酸钠与0.005%TFA混合溶液,配好后调到pH5.0;紫外检测波长196 nm;流速1.0 mL/min;柱温为室温。本方法TTX标准品检出限为1.055 mg/L,血浆中TTX的检出限为0.1055 mg/L。血浆中TTX的线性范围为21.1~211 mg/L,r=0.9989。日内和日间精密度RSD均小于3%。本方该法准确、专属性强,适用于血浆中TTX的浓度测定。     

高效液相色谱-二极管阵列检测法测定L-丙氨酸与L-氨基丙醇

建立了柱前衍生同时测定L丙-氨酸、L氨-基丙醇的高效液相色谱方法。色谱条件为:Shim-pack VP-ODS柱(150 mm×4.6 mm.ID.,5μm);二极管阵列检测器;流动相为甲醇-磷酸盐缓冲溶液(NaH2PO4浓度为0.02 mol/L,pH 4.00 H3PO4调节)(体积比为30∶70),流速为0.5 mL/m in,检测波长为222 nm,柱温为30℃。L丙-氨酸与L氨-基丙醇的线性范围分别为2.30~178 mg/L和9.3~8690 mg/L;检出限分别为1.1 mg/L和1.6 mg/L;日内、日间测定的相对标准偏差(RSD)分别为0.22%、0.46%和0.02%、0.03%。     

高效液相色谱法测定人血浆中的芦氟沙星

 建立了测定人血浆中芦氟沙星质量浓度的高效液相色谱法 ,血浆用二氯甲烷提取 3次 ,以 UltrasphereODS(4.6mm i.d.× 2 5 0 mm)为色谱柱 ,流动相为甲醇 -1 0 mmol/L 溴化四丁铵 -三乙胺 (体积比为 3 2∶ 68∶0 .5 ) ,用磷酸调 p H2 .8,检测波长 2 95 nm,流速为 1 .2 m L/min,以培氟沙星为内标。血浆中芦氟沙星的线性范围为 0 .1～ 1 0 mg/L ,最低检测质量浓度为 0 .0 5 mg/L ,回收率为 99.7% ,日内、日间 RSD分别为 2 .3 3 %和3 .83 %。方法简便、快速、准确 ,适用于人血浆中芦氟沙星质量浓度的测定。     

酸化沉淀蛋白-超快速液相色谱-串联质谱法测定肝损伤大鼠血浆中的头孢呋辛

为了研究二代头孢类新药头孢呋辛赖氨酸在肝损伤大鼠体内的药代动力学过程,建立了采用超快速液相色谱-串联质谱(UFLC-MS/MS)快速测定肝损伤模型大鼠血浆中头孢呋辛含量的方法。血浆样品在酸性条件下用乙腈沉淀蛋白,采用Shim-pack XR-ODS色谱柱(75 mm×3.0 mm, 2.2 μm)为分析柱、乙腈-0.1%甲酸水溶液(40:60, v/v)为流动相、流速为400 μL/min进行色谱分离,采用电喷雾负离子(ESI~)模式电离、多反应监测(MRM)模式进行质谱检测,用于定量分析的离子对分别为m/z 423.2→206.8 (头孢呋辛)和m/z 454.1→238.4 (内标头孢噻肟)。结果表明,大鼠血浆中头孢呋辛的质量浓度在0.01～1 mg/L和1～400 mg/L范围内线性关系良好(r＞0.99),定量限为0.01 mg/L,日内和日间精密度(以相对标准偏差(RSD)计)均小于11.5%,准确度(RE)为~7.1%～2.2%,平均萃取回收率大于83.5%,样品运行时间仅为3.0 min,能够满足生物样品的测定需求。该法简便、快速,已用于肝损伤大鼠静脉注射头孢呋辛赖氨酸的药代动力学预实验研究。     

血浆辅酶Q10的高效液相色谱快速测定

建立了一种简单快速测定血浆辅酶Q10(CoQ10)的反相高效液相色谱方法.血浆经正丙醇萃取,上层清液直接进样分析.色谱柱为Hypersil ODS2 5μm,150 mm ×4.6 mm i.d,以异丙醇-甲醇(体积比19)作流动相,275 nm作检测波长,外标法定量.在0.05～20 mg/L范围内,峰高与质量浓度呈良好的线性关系(r=0.999 8),血浆中辅酶Q10的检出限为0.03 mg/L(S/N=3).该方法简单、快速、精密度高(RSD＜5%),适宜于血浆辅酶Q10含量的检测.     

柱前衍生化高效液相色谱-荧光检测法测定桑叶中的1-脱氧野尻霉素

采用柱前衍生化高效液相色谱-荧光检测法测定了桑叶中的1-脱氧野尻霉素(DNJ)。用0.05 mol/L HCl提取桑叶中的DNJ,采用6-氨基喹啉基-N-羟基琥珀酰亚氨基甲酸酯(AQC)试剂在pH 8.5硼酸盐缓冲液下对DNJ进行衍生化,以0.02 mol/L磷酸二氢钾缓冲液(pH 5.0)-乙腈(体积比为85∶15)为流动相,利用C18色谱柱(5 μm,250 mm×4.6 mm)分离,在激发波长为250 nm、发射波长为395 nm条件下进行荧光检测,DNJ的AQC衍生物与衍生化试剂的水解产物分离良好。方法的线性范围为0.5～25 mg/L,检出限为0.02 mg/L（S/N=3）。实验测得桑叶中DNJ含量为0.12%;回收率为96.1%～98.6%。     

柱前衍生反相高效液相色谱-荧光检测法测定大鼠血浆中的奈替米星

建立了一种简便、灵敏的氯甲酸芴甲酯(FMOC-Cl)柱前衍生反相高效液相色谱-荧光检测血浆中奈替米星的新方法,同时研究了其药代动力学。对色谱条件进行了优化,采用ZORBAX Eclipse XDB-C8柱(150 mm×4.6 mm,5 μm),流动相为乙腈-水(体积比为85:15),流速为1.0 mL/min,荧光检测激发波长为265 nm,发射波长为315 nm,得到奈替米星的平均加标回收率为96.62%～100.84%(n=3),对奈替米星检测的线性范围为0.045～8.88 mg/L,相关系数为0.9993,方法的日内与日间精密度分别低于3%与3.5%,最低检出限(S/N=3)与定量限(以3倍检出限计)分别为0.01和0.03 mg/L。方法简便、快速、灵敏,样品用量少(30 μL奈替米星血浆溶液已能满足该药含量的测定以及药物代谢的研究),为大鼠体内奈替米星的药代动力学研究提供了可靠的分析手段。     

高效液相色谱法测定奶粉和液态奶中的三聚氰胺

建立了奶粉和液态奶中三聚氰胺的高效液相色谱（HPLC）检测方法。经阳离子交换固相萃取柱净化后的样品采用HPLC测定。优化的色谱条件：C18柱（4.6 mm ×200 mm,5 μm）,流动相为乙腈-0.01 mol/L庚烷磺酸钠（pH 3.3）（体积比为10∶90）,流速为1.0 mL/min,检测波长为236 nm,柱温为40 ℃,进样量为20 μL。方法的线性范围为1~500 mg/L,检出限为0.2 mg/kg （S/N=3）,定量限为1 mg/kg （S/N=15）,回收率为81.4%~83.7%,相对标准偏差为3.3%~8.5%（n=6）。     

超高效液相色谱-三重四极杆质谱法测定血浆和尿液中马桑亭和马桑宁

建立了超高效液相色谱-三重四极杆质谱联用技术测定血浆和尿液中马桑中毒标志物马桑亭和马桑宁的方法。血浆和尿液样品经固相支持液液萃取法提取净化后，溶于15%（v/v）甲醇水溶液中，以Cortecs C₁₈色谱柱（100 mm×2.1 mm，1.6 μm）作为分析柱进行分离，电喷雾负离子多反应监测（MRM）模式下检测，以氟苯尼考作为内标物，基质工作曲线内标法定量。血浆和尿液中马桑亭和马桑宁的平均加标回收率为86.2%~110%，相对标准偏差为5.1%~14.6%（n=6），血浆中马桑亭和马桑宁的检出限（S/N=3）分别为0.01 μg/L和0.1 μg/L，尿液中马桑亭和马桑宁的检出限分别为0.03 μg/L和0.3 μg/L。本法简单、灵敏、准确，可用于血浆和尿液中马桑亭和马桑宁的中毒检测。     

固相萃取高效液相色谱法测定人血浆中的巴洛沙星

建立了测定人血浆中巴洛沙星含量的固相萃取高效液相色谱紫外检测方法。采用pH 4.5的磷酸盐缓冲液并经Waters Oasis HLB固相萃取小柱对血浆样品进行预处理。以环丙沙星作内标,以十二烷基磺酸钠溶液(取560 mL水加入3.2 g十二烷基磺酸钠)-乙腈(体积比为56∶44,用磷酸调pH至3.0)为流动相,在Diamonsil C18色谱柱(150 mm×4.6 mm,5 μm)上进行分离。方法的线性范围为25～3200 μg/L,线性关系良好(r=0.9996);巴洛沙星的检出限（S/N≥3）为5 μg/L;方法的准确度为100.3%～103.8%;平均提取回收率为57.5%～77.0%,相对标准偏差小于5.1%。方法准确、灵敏,可满足血药浓度监测和药代动力学参数测试的需求。     

HPLC analysis and pharmacokinetics of icariin in rats

As a prerequisite to the determination of pharmacokinetic parameters of icariin in rats, an HPLC method using UV detection was developed and validated. Icariin and the internal standard, quercetin, were extracted from plasma samples using ethyl acetate after acidification with 0.05 mol/L NaH2PO4 solution (pH 5.0). Chromatographic separation was achieved on an Agilent XDB Cls column (250 x 4.6 mm id, 5 microm) equipped with a Shim-pack GVP-ODS C18 guard column (10 x 4.6 mm id, 5 microm) using a mobile phase of ACN/water/acetic acid (31:69:0.4 v/v/v) at a flow rate of 1.0 mL/ min. Detection was at 277 nm. The calibration curve was linear from 0.05 to 100.0 microg/mL with 0.05 microg/mL as the lower LOQ (LLOQ) in plasma. The intra- and interday precisions in terms of RSD were lower than 5.7 and 7.8% in rat plasma, respectively. The accuracy in terms of relative error (RE) ranged from -1.6 to 3.2%. The extraction recoveries of icariin and quercetin were 87.6 and 80.1%, respectively. The main pharmacokinetic parameters for rats were determined after a single intravenous administration of 10 mg/kg icariin: t1/2, 0.562 +/- 0.200 h; AUC0-infinity, 8.73 +/- 2.23 microg x h/mL; CLToT, 20.10 +/- 5.80 L/kg x h; Vz, 1.037 +/- 0.631 L/kg; MRT0-infinity, 0.134 +/- 0.040 h; and Vss, 0.170 +/- 0.097 L/kg.     

高效液相色谱-质谱法测定比格犬血浆中的艾普拉唑及其药代动力学

运用高效液相色谱-电喷雾质谱(HPLC-ESI-MS)技术,建立了快速、简单、灵敏的比格犬静脉滴注艾普拉唑钠盐后血药浓度的检测方法。血浆样品采用蛋白沉淀法,以丁螺环酮作为内标,色谱柱为Teknokroma Kromasil C18(100 mm×2.1 mm, 5 μm),流动相为水-甲醇-乙腈(69:8:23, v/v/v)(含0.1%的甲酸),流速0.2 mL/min,采用电喷雾(ESI)离子源以正离子方式检测。绘制血药浓度-时间曲线,并采用DAS 2.0计算药代动力学参数。方法学实验结果表明内源性杂质不干扰艾普拉唑和内标的测定,线性范围为5～10000 μg/L (r=0.994),最低定量限为5 μg/L,精密度和准确度均符合生物样品测定的要求。低、中、高3个浓度的绝对回收率在106%左右,基质效应小于142.0%,表明该方法适合比格犬血浆中艾普拉唑浓度的测定及药代动力学研究。比格犬静脉滴注艾普拉唑钠盐3个剂量(0.2 mg/kg、0.8 mg/kg和3.2 mg/kg)后的药-时曲线下面积(AUC(0~∞))分别为(2.4×104±3×103)、(8.8×104±1.6×104)和(5.4×105±8×104) μg/L•min,呈线性药物代谢动力学过程。     

Determination and pharmacokinetic study of syringin and chlorogenic acid in rat plasma after administration of Aidi lyophilizer

A high-performance liquid chromatographic (HPLC) method was developed for the first time to simultaneously quantify syringin and chlorogenic acid in rat plasma using wavelength-transfer technology. The analysis was performed on a Diamonsil C(18) column (200 x 4.6 mm i.d., 5 microm particle size) with isocratic mobile phase consisting of acetonitrile-0.05% phosphoric acid (12:88, v/v). The linear ranges were 0.20-10 and 0.25-30 microg/mL, respectively. The lower limits of quantification were 0.20 and 0.25 microg/mL, respectively. The method was shown to be reproducible and reliable with intraday precision below 8.5 and 6.1%, interday precision below 7.1 and 5.5%, accuracy within +/-7.1 and +/-8.6%, and mean extraction recovery excess of 92.1 and 80.9%, respectively, which were all calculated from the blank plasma sample spiked with syringin and chlorogenic acid at three concentrations of 0.20, 1.0 and 6.0 microg/mL for syringin and 0.25, 2.0 and 20 microg/mL for chlorogenic acid. This method was validated for specificity, accuracy and precision and was successfully applied to the pharmacokinetic study of syringin and chlorogenic acid in rat plasma after intravenous administration of Aidi lyophilizer.     

Determination and pharmacokinetic study of chlorogenic acid in rat dosed with Yin-Huang granules by RP-HPLC

A reversed-phase high-performance liquid chromatographic (RP-HPLC) method was described for the determination of chlorogenic acid (CGA) in rat plasma using protocatechuic acid as internal standard (IS). CGA in plasma was extracted with acetonitrile, which also acted as deproteinization agent. Chromatographic separation was performed on a Kromasil C18 column with methanol-0.2 m acetic acid (pH 3.0, 25:75, v/v) as mobile phase at a flow-rate of 1.0 mL/min with an operating temperature of 30 degrees C and UV detection at 300 nm. The standard curve was found to be linear over the concentration ranges of 0.4-2.5 microg/mL and 2.5-40 microg/mL, and the limit of quantification (LOQ) was 0.4 microg/mL. The analytical precision and accuracy were validated by relative standard deviation (RSD) and relative error, which were in ranges 3.14-10.78% and -2.20-5.00%, respectively. The average recovery of CGA was 87.59%. The method was successfully applied to the pharmacokinetic study of CGA in Yin-Huang granules.     

HPLC determination and pharmacokinetics of osthole in rat plasma after oral administration of Fructus Cnidii extract

A simple and sensitive high-performance liquid chromatographic (HPLC) method is developed for the determination of osthole in rat plasma and applied to a pharmacokinetic study in rats after administration of Fructus Cnidii extract. After addition of fluocinonide as an internal standard, plasma samples are extracted with diethyl ether. HPLC analysis of the extracts is performed on a Hypersil ODS2 analytical column, using methanol-0.4% acetic acid (65:35, v/v) as the mobile phase. The UV detector is set at 322 nm. The standard curve is linear over the range 0.0520-5.20 microg/mL (r = 0.9979). The mean extraction recoveries of osthole at three concentrations were 81.0%, 91.2%, and 90.7%, respectively. The intra- and interday precisions have relative standard deviations from 1.9% to 4.9%. The limit of quantitation is 0.0520 microg/mL. The HPLC method developed can easily be applied to the determination and pharmacokinetic study of osthole in rat plasma after the animals are given the Fructus Cnidii extract. The plasma concentration of osthole from six rats showed a Cmax of 0.776 +/- 0.069 microg/mL at Tmax of 1.0 +/- 0.3 h.     

Development of an HPLC method for the determination of salidroside in beagle dog plasma after administration of salidroside injection: application to a pharmacokinetics study

A simple RP-HPLC method was established for the determination of salidroside in dog plasma. Salidroside is one of the most active ingredients of Rhodiola L. The method had within-run precision values in the range of +/- 2.3 to +/- 9.1% (n = 5) and between-run precision in the range of +/- 3.2 to +/- 9.8%. A simple protein precipitation for salidroside extraction was processed using ACN at precipitant-to-plasma volume ratio (P-P ratio) of 3:2. The extraction recoveries of salidroside at seven concentrations were higher than 63.2%. There was a linear relationship between chromatographic area and concentration over the range of 0.83-520 microg/mL for salidroside in plasma (R = 0.9926). The LOQ (S/N = 10) of the method was 0.83 microg/mL. The method was applied in a study of the pharmacokinetics of salidroside injection in six beagle dogs. The major pharmacokinetic parameters of C(max), AUC(0-24), AUC(0-infinity), and t(1/2) of salidroside in beagle dogs after i.v. administration of a single 75 mg/kg (5 mL/kg) dose were 96.16 +/- 8.59 microg/mL, 180.3 +/- 30.6 microg h/mL, 189.3 +/- 32.1 microg h/mL, and 2.006 +/- 0.615 h, respectively.     

HPLC determination of andrographolide in rat whole blood: study on the pharmacokinetics of andrographolide incorporated in liposomes and tablets

A sensitive and simple high-performance liquid chromatographic method with UV detection was developed and validated for the determination of andrographolide in rat whole blood. Carbamazepine was employed as internal standard and the blood sample was extracted with chloroform. Chromatographic separations were achieved on a Chromasil ODS column (250 x 4.6 mm, 5 microm). The mobile phase was consisted of methanol-water (52:48, v/v) and delivered at 0.8 mL/min. The detection wavelength was set at 225 nm. The calibration curve had a good linearity in the range 0.053-530 microg/mL in rat whole blood with its correlation coefficient being 0.996. The extraction recovery of andrographolide was ranged from 65.7 to 72.6%. The intra-day and inter-days repeatabilities were below 4.2% in terms of the percentage of relative standard deviation (RSD). The method was used to provide data on the pharmacokinetics of the drug in rats. The data obtained was processed using the 3P87 pharmacokinetic program. The results showed that the disposition of andrographolide after intravenous administration of liposomal andrographolide conformed to a two-compartment open model with alpha = 4.75 x 10(-2) +/- 2.41 x 10(-3) min(-1), beta = 3.16 x 10(-3) +/- 1.58 x 10(-4) min(-1), V(c) = 174.67 +/- 13.97 mL, k(21) = 1.60 x 10(-2) +/- 8.12 x 10(-4) min(-1), k(10) = 9.38 x 10(-3) +/- 5.62 x 10(-4) min(-1), k(12) = 2.53 x 10(-2) +/- 1.27 x 10(-3) min(-1) and AUC(0-infinity) = 1525.47 +/- 92.35 microg min/mL. For the intragastric administration of andrographolide tablets, the disposition of andrographolide followed a one-compartment open model with k(e) = 6.78 x 10(-3) +/- 3.53 x 10(-4) min(-1), k(a) = 3.69 x 10(-2) +/- 4.68 x 10(-3) min(-1), T(max) = 59.69 +/- 3.61 min, C(max) = 1.62 +/- 0.11 microg/mL, V(c) = 1056.90 +/- 83.42 mL, AUC(0-infinity) = 348.75 +/- 24.41 microg min/mL.     

A simple HPLC method for the quantification of mizoribine in human serum: pharmacokinetic applications

A simple high-performance liquid chromatographic (HPLC) method was developed and validated for the quantification of mizoribine in human serum. After the addition of 70% perchloric acid and 3-methylxanthine (50 microg/mL, internal standard) to human serum, the samples were mixed and centrifuged at 12,000 rpm (1432 g) for 10 min. The supernatant was injected onto a C(18) column eluted with a mobile phase of 20 mm Na2HPO4 and methanol (93:7, v/v, pH 3) containing 0.04% octanesulfonic acid and detected utilizing an ultraviolet detector at 275 nm. The linear calibration curve was obtained in the concentration range of 0.1-4.0 microg/mL and the lower limit of quantification was 0.1 microg/mL. This method was validated with selectivity, linearity, precision and accuracy. In addition, the method was successfully applied to estimate the pharmacokinetic parameters of mizoribine in Korean subjects following an oral administration of 100 mg mizoribine (two Bredinine 50 mg tablets). The maximum serum concentration (C(max)) of 2.30 +/- 0.83 microg/mL was reached 2.27 +/- 0.66 h after an oral dose. The mean AUC(0-12 h) and the elimination half-life (t(1/2)) were 13.2 +/- 4.79 microg h/mL and 3.10 +/- 0.74 h, respectively.     

HPLC determination and pharmacokinetics of chlorogenic acid in rabbit plasma after an oral dose of Flos Lonicerae extract

A simple and sensitive high-performance liquid chromatography (HPLC) method has been developed for the determination of chlorogenic acid (3-O-caffeoyl-D-quinic acid) in plasma and applied to its pharmacokinetic study in rabbits after administration of Flos Lonicerae extract. Plasma samples are extracted with methanol. HPLC analysis of the extracts is performed on a C(18) reversed-phase column using acetonitrile-0.2% phosphate buffer (11:89, v/v) as the mobile phase. The UV detector is set at 327 nm. The standard curves are linear in the range 0.0500-1.00 microg/mL (r = 0.9987). The mean extraction recovery of 85.1% is obtained for chlorogenic acid. The interday precision (relative standard deviation) ranges from 5.0% to 7.5%, and the intraday precision is better than 9.0%. The limit of quantitation is 0.0500 microg/mL. The plasma concentration of chlorogenic acid shows a C(max) of 0.839 +/- 0.35 microg/mL at 34.7 +/- 1.1 min and a second one of 0.367 +/- 0.16 microg/mL at 273.4 +/- 39.6 min.     

High performance liquid chromatographic method for the determination and pharmacokinetic studies of oxyresveratrol and resveratrol in rat plasma after oral administration of Smilax china extract

A sensitive and simple HPLC method has been developed and validated for the determination of oxyresveratrol (trans-2,4,3',5'-tetrahydroxystilbene, OXY) and resveratrol (trans-3,5,4'-trihydroxystilbene, RES) in rat plasma. The plasma samples were extracted with ethyl acetate and analyzed using HPLC on an Aglient Zorbax SB-C(18) column (250 x 4.6 mm, 5 microm) at a wavelength 320 nm, with a linear gradient of (A) acetonitrile and (B) 0.5% aqueous acetic acid (v/v), at a flow rate of 1.0 mL/min. The method was linear over the range of 0.1265-25.3 microg/mL for OXY and 0.117-23.4 microg/mL for RES. The extraction recovery for OXY, RES and internal standard ranged from 71.1 to 88.3%. The intra- and inter-day precisions were better than 10%, and the accuracy ranged from 89 to 108%. The validated method was used to study the pharmacokinetic profiles of OXY and RES in rat plasma after oral administration of Smilax china root extract.