高效液相色谱法测定康得灵胶囊中黄芩苷的含量

目的 建立康得灵胶囊中黄芩苷的HPLC测定方法。方法 色谱柱为Agilent Tc-C₁₈色谱柱（4.6 mm×250 mm,5 μm）,柱温为30 ℃;流动相为乙腈-0.5‰磷酸溶液（26：74）,流速为1.0 ml/min,检测波长265 nm。结果 黄芩苷保留时间约为16 min。以峰面积（Y）对进样浓度（X, μg/ml）线性回归,得回归方程：Y=22 114.67 X－112 836.7,r=0.998 8,线性范围5.410~108.2 μg/ml。平均加样回收率为98.78%,RSD为0.74%。结论 本方法操作简便,测定结果准确可靠,可用于康得灵胶囊中黄芩苷的含量测定。     

高效液相色谱法测定人血浆中阿克他利的浓度及其药动学研究

目的 建立人血浆中阿克他利浓度的高效液相色谱检测方法。 方法 血浆样品采用液-液萃取的方法,采用Dikma C₁₈柱分离,以水(含1.2%乙酸)-甲醇 (50:50, V/V) 的混合溶液为流动相,检测波长为245 nm,流速为1.0 ml/min,柱温30 ℃。 结果 建立的人血浆内阿克他利测定方法线性范围为75~4 000 ng/ml,定量下限可达75 ng/ml。日内、日间精密度(RSD)均小于10%,方法准确度大于90%,提取回收率大于88%。 结论 该方法快速、灵敏、专属,适用于人血浆中阿克他利浓度的测定。     

阿糖胞苷在大鼠体内转化为阿糖尿苷的药动学研究

目的 建立测定大鼠血浆中阿糖尿苷的LC-MS/MS方法,用于大鼠尾iv注射用盐酸阿糖胞苷后阿糖尿苷在体内的药动学研究。方法 采用LC-MS/MS法。ACQUITY UPLC BEH C₁₈色谱柱（50 mm×2.1 mm,1.7 μm）;流动相：水–乙腈,梯度洗脱;体积流量：0.2 mL/min;柱温：40 ℃;进样量：5 μL。离子源：ESI源;扫描方式：多反应监测（MRM）方式,扫描时间为0.1 s;毛细管电压：2.5 kV;锥孔电压：26 V;离子源温度：110 ℃;去溶剂气温度：350 ℃;去溶剂气流量：500 L/h;锥孔气流量：50 L/h。采用回归方程计算血浆中阿糖尿苷。SD大鼠尾iv注射用盐酸阿糖胞苷,制备血药质量浓度–时间曲线,计算药动学参数。结果 阿糖尿苷在1.0～1 000 ng/mL线性关系良好,日内、日间RSD值均小于15%,准确度在±15%,平均提取回收率在90%以上,基质效应在97.3%,稳定性良好。药动学参数：t_max是1.0 h,C_max是134.2 ng/mL,AUC_0-t是2 316.0 ng•h/mL,t_1/2是4.3 h。结论 该方法适合大鼠尾iv阿糖胞苷后阿糖尿苷在体内的药动学研究。     

应用高效液相色谱法测定猫人参中积雪草酸的含量

目的 建立猫人参药材中积雪草酸的含量测定方法。 方法 采用高效液相色谱法,色谱柱为Agilent HC-C₁₈柱(4.6 mm×250 mm,5.0 μm),流动相为乙腈(30 mmol/L)-醋酸铵水溶液(35:65),流速1.0 ml/min,柱温25 ℃,紫外检测波长210 nm,进样量25 μl,运行时间35 min。 结果 积雪草酸与周围干扰峰达到基线分离,线性范围25.3~506.0 μg/ml, r=0.999 6;日内及日间精密度均<3% (n=5),平均回收率为99.4%(RSD=1.9%, n=6),猫人参中积雪草酸的含量为0.751 mg/g。 结论 该法简便快捷、测定结果准确、实用性强,可用于猫人参药材中积雪草酸的含量测定。     

高效液相色谱法测定人血浆中盐酸氨溴索的浓度

目的 建立HPLC测定人血浆中盐酸氨溴索含量的方法。 方法 选用Diamonsil-C₁₈柱(200 mm×4.6 mm,5μm),流动相:甲醇-0.01 mol/L磷酸盐缓冲液(pH 7.0)-四氢呋喃(700:320:25,v/v/v),流速:1.2 ml/min,检测波长:242 nm,柱温:室温。 结果 该方法低、中、高3种浓度的提取回收率为88.2％ ~104.8％ ,日内、日间相对标准差为2.7％~5.3％(n=6),在10.2~610.8 ng/ml范     

复方虎杖酊的质量标准研究

目的 建立复方虎杖酊的质量控制标准。方法 采用薄层色谱法对复方虎杖酊中虎杖、蜂房和冰片进行定性鉴别;采用HPLC法测定虎杖中虎杖苷的含量,色谱柱为Agilent Eclipse XDB-C₁₈柱（4.6 mm×250 mm,5 μm）,流动相：乙腈-水（23：77）,检测波长306 nm,流速1.0 ml/min,柱温40℃。结果 复方虎杖酊中虎杖、蜂房和冰片的薄层色谱图斑点明显,阴性对照无干扰;虎杖苷的定量测定在8.25~66.0 μg/ml范围内与峰面积呈良好的线性关系（r=0.999 9）,平均回收率为100.1%,RSD为0.2%。结论 本实验建立的方法快速简便,准确可靠,可作为该制剂的质量控制方法。     

高效液相色谱法测定气管炎颗粒中黄芩苷的含量

目的 建立高效液相色谱法测定气管炎颗粒中黄芩苷的含量。 方法 采用HPLC法,色谱柱:ZORBAX SB-C₁₈柱(4.6 mm×150 mm,5 μm),以甲醇-0.1%磷酸(47:53)为流动相,流速为1 ml/min,检测波长为280 nm。 结果 黄芩苷在6.63~210 μg/ml范围内呈良好线性关系(r=0.999 8,n=6),平均加样回收率为98.16%,RSD为1.89%。 结论 本法简便、灵敏、准确、重复性好,结果可靠,可有效地控制气管炎颗粒的质量。     

大鼠一次性灌服酸枣仁提取物后棘苷的药代动力学研究

目的用反相高效液相色谱法,以磺胺甲唑为内标,对一次性ig酸枣仁提取物后的大鼠血浆中棘苷进行药代动力学研究。方法血浆样品经乙腈沉淀蛋白后,于50 ℃氮气流下吹干,残渣用流动相溶解后进行分析。色谱条件为色谱柱:Hypersil C₁₈柱,200 mm×4.6 mm ID,5 μm;流动相:乙腈-水-冰醋酸(15∶85∶1);流速:0.7 mL·min^-1;检测波长:334 nm;柱温:35 ℃。结果血浆中棘苷在18.1～903.5 μg·L^-1成良好线性关系(R²≥0.995)。平均回收率为94.5%,日内、日间精密度RSD均小于9.0%。该法定量限为18.1 μg·L^-1,血浆样品在-20 ℃可稳定保存。结论该法简便、灵敏、准确,可用于大鼠一次性灌服酸枣仁提取物后血浆中棘苷的浓度测定及其药代动力学研究。     

LC-MS/MS测定人血浆中多黏菌素B浓度及其应用

目的 建立测定人血浆中多黏菌素B总浓度的液相色谱串联质谱(LC-MS/MS)方法，并应用于临床多黏菌素B的治疗药物监测。方法 采用Welch Ultimate LP-C₁₈(100mm×2.1mm，5μm)色谱柱对多黏菌素B的主要成分进行色谱分离，建立时间程序为4.0min的LC-MS/MS法，以乙腈为沉淀剂进行蛋白沉淀的样品前处理，用正离子扫描以及多反应监测模式进行分析物测定。结果 多黏菌素B的总浓度线性范围为100~10000ng·mL^-1，各主要成分线性系数均>0.996，批内和批间精密度RSD均≤6%，准确度为91.88%~114.02%，测定3例使用多黏菌素B患者的体内暴露量，AUC_0-24h分别为39.73，64.15，107.75mg·L^-1·h^-1。结论 本研究建立了一种稳健的测定人血浆中多黏菌素B总浓度的LC-MS/MS方法，灵敏度和特异性好，并成功用于临床上多黏菌素B的治疗药物监测。     

用HPLC法测定红茴香注射液中槲皮苷的含量

目的 建立高效液相色谱法测定红茴香注射液中槲皮苷的含量。方法 色谱柱为Dikma C₁₈(250 mm×4.6 mm,5 μm),以乙腈-0.1%磷酸水溶液（25∶75）为流动相,流速为1.0 ml/min,柱温30 ℃,检测波长为256 nm。结果 槲皮苷在0.215～3.225 μg范围内呈良好线性关系,相关系数为0.999 6,该法的平均回收率为99.39%,RSD为0.82%（n=6）,重复性为1.194 mg/ml,RSD为0.40%。结论 3批红茴香注射液的槲皮苷平均含量结果为1.191 mg/ml,该法简便、快速、准确, 可作为测定红茴香注射液中槲皮苷含量的方法。     

Development and validation of a LC-MS/MS method with electrospray ionization for determination of LASSBio-579 in rat plasma

A simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of LASSBio-579 in plasma rat, using fluconazole as internal standard. Analyses were performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water (80:20, v/v), containing 0.4mM ammonium hydroxide and 0.2 mM acetic acid at a flow rate of 1.0 ml/min (split ratio 1:5). A Micromass triple quadrupole mass spectrometer, equipped with an electrospray ionization interface, operated in the positive mode. Plasma samples were deproteinized with acetonitrile (1:2) and 50 microl of the supernatant were injected into the system. The retention times of LASSBio-579 and IS were approximately 4.7 and 2.4 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 30-2000 ng/ml with determination coefficient >0.98. The lower limit of quantification was 30 ng/ml. The accuracy of method was within 15%. Intra- and inter-day relative standard deviations were less or equal to 13.5% and 6.4%, respectively. The applicability of the LC-MS/MS method for pharmacokinetic studies was tested using plasma samples obtained after intraperitoneal administration of LASSBio-579 to male Wistar rats. No interference from endogenous substances was observed, showing the specificity of the method developed. The reported method can provide the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of LASSBio-579 in pre-clinical pharmacokinetic studies.     

LC-MS determination and pharmacokinetic studies of ursolic acid in rat plasma after administration of the traditional chinese medicinal preparation Lu-Ying extract

Sambucus chinensis L. is a native perennial herb distributed throughout China. In traditional Chinese medicine (TCM), this herb is known as Lu-Ying. Ursolic acid is the major effective constituent of Lu-Ying. A rapid, sensitive, and accurate liquid chromatography-mass spectrometry (LC-MS) method for the determination of ursolic acid in rat plasma was developed and validated. Plasma samples taken from rats that had received Lu-Ying extract orally were acidified with acetic acid and then extracted with a mixture of hexane-dichloromethane-2-propanol (20:10:1, v/v/v). Separation of ursolic acid was accomplished on a C(18) column interfaced with a single quadrupole mass spectrometer. The mobile phase consisting of methanol and water (95:5, v/v) was delivered at a flow rate of 1.0 ml/min. Atmospheric pressure chemical ionization was operated in negative-ion mode. Using selected ion-monitoring mode, the deprotonated molecules [M-H](-) at m/z 455 and 469 were used to quantify ursolic acid and glycyrrhetic acid (internal standard), respectively. The assay was shown to be linear over the range of 10-1000 ng/ml (r> or =0.9960) with a lower limit of quantification of 10 ng/ml. The method was shown to be reproducible and reliable with intraday precision below 7.8%, interday precision below 8.1%, accuracy within +/-4.3%, and mean extraction recovery excess of 83.6%, which were all calculated from the blank plasma sample spiked with ursolic acid at three concentrations of 20, 200, and 800 ng/ml. The LC-MS method has been successfully applied to pharmacokinetic studies of ursolic acid after oral administration of Lu-Ying ethanolic extract (at a dose containing 80.32 mg/kg ursolic acid) to rats. The main pharmacokinetic parameters were: t(1/2), 4.3 h; K(e), 0.16 1/h; t(max), 1.0 h; C(max), 294.8 ng/ml; AUC(0-t) and AUC(0-infinity), 1007.1 ng.h/ml and 1175.3 ng.h/ml, respectively.     

Pharmacokinetic study of the novel, synthetic trioxane antimalarial compound 97-78 in rats using an LC-MS/MS method for quantification

The present study has been designed to investigate the pharmacokinetic parameters of the novel trioxane antimalarial 97-78 (US Patent 6316493 B1, 2001) in male and female rats after single oral and intravenous administration. The pharmacokinetic profile of 97-78 was investigated in the form of its completely converted metabolite 97-63 after dose administration. Quantification of metabolite 97-63 in rat plasma was achieved using a simple and rapid LC-MS/MS method. The LC-MS/MS method has been validated in terms of accuracy, precision, sensitivity and recovery for metabolite 97-63 in rat plasma. The intra- and interday accuracy (% bias) and precision (% RSD) values of the assay were less than 10% for metabolite 97-63. The chromatographic run time was 4.0 min and the weighted (1/x2) calibration curves were linear over the range 1.56-200 ng/ml. This method was successfully applied for analysis of pharmacokinetic study samples. Maximum plasma concentrations of 97-63 at 47 mg/kg oral administration in male and female rats were 1986.6 ng/ml and 4086.7 ng/ml at time (Tmax) 0.92 h and 0.58 h, respectively. The area under the curve (AUC(0-infinity)), elimination half-life (t(1/2) beta) and mean residence time (MRT) were 4669.98 ng x h/ml, 2.8 h and 4.2 h in male and 11786.0 ng x h/ml, 4.52 h and 4.32 h in female rats respectively. After single oral and intravenous administration of 97-78 to male and female rats significant differences were observed in pharmacokinetic parameters (AUC and t (1/2) beta) for metabolite 97-63.     

A rapid and sensitive LC-MS/MS assay to quantify yonkenafil in rat plasma with application to preclinical pharmacokinetics studies

A novel method for the quantitation of yonkenafil, a new synthetic phosphodiesterase V inhibitor, in rat plasma using high-performance liquid chromatography/tandem mass spectrometry (LC-MS/MS) has been developed. The analyte and internal standard (diazepam) were extracted from plasma (100 microl) by liquid-liquid extraction and separated on a C18 column using 10mM ammonium acetate buffer: methanol (15:85, v/v) as mobile phase in a run time of 3.0 min. The detector was a Q-trap mass spectrometer with an ESI interface operating in the multiple reaction monitoring (MRM) mode. The assay was linear over the concentration range 1.0-1000 ng/ml with a limit of detection of 0.20 ng/ml. Intra- and inter-day precision (as relative standard deviation) were both within 8.45% with good accuracy. The method was successfully applied to a preclinical pharmacokinetic study of yonkenafil in rat after sublingual, oral and intravenous administration. The results demonstrate that the sublingual route gives a higher bioavailablity than the oral route and may represent a useful alternative route of yonkenafil administration.     

LC-MS/MS法测定大鼠血浆中吴茱萸碱的浓度及其药代动力学研究

目的：建立用于测定吴茱萸碱血药浓度的液相色谱-串联质谱联用分析方法,并研究吴茱萸碱在大鼠体内的药代动力学。方法：6只大鼠灌胃给药吴茱萸碱100mg/kg,眼底取血,LC-MS/MS法测定血药浓度,并用DAS药代动力学程序拟合计算药代动力学参数。结果：吴茱萸碱浓度在0.2～50ng/mL内,线性关系良好（r^2=0.9997）。提取回收率96.12%～99.46%,日内、日间RSD分别为4.61%～13.51%和5.65%～11.49%。主要药代动力学参数为：Cmax=（5.3±1.5）ng/mL;tmax=（22±8）min;t1/2=（451±176）min。结论：建立的LC-MS/MS方法专属性强,灵敏度高,可用于吴茱萸碱的体内定量分析。     

Validation of rapid and simple LC-MS/MS method for determination of voriconazole in rat plasma

A rapid, simple and sensitive LC-MS/MS analytical method was developed and validated for the determination of voriconazole (VRC) in rat plasma, using ketoconazole as internal standard (IS). Analysis was performed on a Shimadzu HPLC system using a Shimadzu C18 column and isocratic elution with acetonitrile-water-formic acid (60:40:0.05, v/v/v), at a flow of 1.0 mL/min (split ratio 1:5), and a mass spectrometer Micromass, equipped with a double quadrupole and an electrospray ionization interface, operated in a positive mode. Plasma samples were deproteinized with methanol (1:2) and 30 microL of the supernatant was injected into the system. The retention times of VRC and IS were approximately 3.3 and 2.7 min, respectively. Calibration curves in spiked plasma were linear over the concentration range of 50-2500 ng/mL with determination coefficient >0.98. The lower limit of quantification was 50 ng/mL. The accuracy of the method was within 5%. Intra- and inter-day relative standard deviations were less or equal to 12.5 and 7.7%, respectively. The applicability of the LC-MS-MS method for pharmacokinetic studies was tested using plasma samples obtained after intravenous administration of VRC to male Wistar rats. The reported method provided the necessary sensitivity, linearity, precision, accuracy, and specificity to allow the determination of VRC in pre-clinical pharmacokinetic studies.     

Determination of deoxyschizandrin in rat plasma by LC-MS

A simple, rapid and sensitive LC-MS method was developed for quantification of deoxyschizandrin in rat plasma. A 50 miccrol plasma sample was extracted by ether and performed on Elite Hypersil C(18) column (200 mm x 4.6 mm, 5 microm) with the mobile phase of methanol-water (84:16, v/v) in a run time of 6.5 min. The analyte was monitored with positive atmospheric pressure chemical ionization (APCI) by selected ion monitoring (SIM) mode. A good linear relationship was obtained over the range of 1.0-50.0 ng/ml and the validated method was successfully applied for the pharmacokinetic studies of deoxyschizandrin in rat. After oral administration of 4 mg/kg deoxyschizandrin and Schisandra extract which contained the same dose of deoxyschizandrin to male rats, the C(max) of deoxyschizandrin were 15.8+/-3.1 and 34.3+/-16.8 ng/ml, T(max) were 0.51+/-0.13 and 3.83+/-1.83 h, T(1/2) were 5.3+/-2.2 and 6.5+/-3.4h.     

Simultaneous rapid quantification of ginsenoside Rg1 and its secondary glycoside Rh1 and aglycone protopanaxatriol in rat plasma by liquid chromatography-mass spectrometry after solid-phase extraction

Ginseng saponins isolated from ginseng, have been regarded as the principal constituents responsible for the biological activities. The aim of this study was to set up a liquid chromatography-mass spectrometry (LC-MS) method for simultaneously determine the concentration of Ginsenoside Rg1 and its secondary glycoside Rh1 and aglycone protopanaxatriol (PPT) in rat plasma so as to study the pharmacokinetics of Rg(1) after intraveneous (i.v.) and intragastric gavage (i.g.) administration. One hundred microliters or 1.0 ml of rat plasma samples from i.v. or i.g. treated rats were used respectively for analysis. After solid-phase extraction (SPE) and high performance liquid chromatography (HPLC) separation, the chloride adduct anions [M+Cl]- of Rg1, Rh1 and PPT were analyzed by LC-MS in selected ions monitoring (SIM) mode. Rg(1) could be determined by this LC-MS method over the ranges of 1.56-250 ng/ml and 250-20,000 ng/ml with the correlation coefficients of 0.999 and 0.9998, respectively. The detection limits (LOD) of this method was 20 pg (S/N>3) for Rg1, 100 pg for Rh1 and 10 pg for PPT. Chromatographic separation was achieved in less than 8 mins. The method has been used for the pharmacokinetic study of Rg1 in rats.     

A rapid HPLC/ESI-MS method for the quantitative determination of oridonin in rat plasma

A rapid and accurate method using liquid chromatography with electrospray ionization mass spectrometric detection (HPLC/ESI-MS) was developed and validated for the determination of oridonin in rat plasma. The analytes were extracted with ethyl acetate-n-butyl alcohol (100:2, v/v) after spiking the samples with ethyl hydroxybenzoate (internal standard). The separation was carried out on a Diamon-sil C18 column with an isocratic mobile phase consisting of methanol-water (80:20, v/v) at a flow rate of 1.0 ml/min. The lower limit of quantification (LLOQ) of the method was 10 ng/ml and the linear range was 10-4000 ng/ml. The intra-day and inter-day accuracy and precision of the assay were less than 9%. This method has been applied successfully to a preliminary pharmacokinetic study involving the intravenous administration of oridonin to rats.     

LC-MS-MS法测定血浆中多西他赛和紫杉醇浓度及在乳腺癌患者中的应用

目的：建立一种快速、灵敏的液相色谱－串联质谱（LC-MS/MS）法检测乳腺癌患者血浆中多西他赛、紫杉醇的浓度。方法：多西他赛和紫杉醇互为内标,血浆样品100μL加入1 mL叔丁基甲醚萃取,分离有机相,以氮气吹干后流动相复溶进样。色谱柱为Agilent Eclipse XDB-C18（2.1 mm×100 mm,3.5μm）,流动相为0.4%甲酸水溶液-0.4%甲酸乙腈溶液（20∶80,v/v）,流速0.3 mL·min-1,柱温为40℃。采用多反应监测（MRM）进行定量,电喷雾电离源（ESI）正离子方式进行检测,多西他赛与紫杉醇用于定量分析的检测离子对分别为m/z 808.5→m/z 527.2和m/z 854.3→m/z 569.4。结果：多西他赛和紫杉醇的线性范围分别为5.0~1000 ng·mL-1和1.0～500 ng· mL-1,最低检测浓度分别为5.0 ng·mL-1和1.0 ng·mL-1。两药低、中、高三个浓度的批内和批间RSD均<15%,平均提取回收率分别为65.9%~84.3%和90.4%~106.5%。结论：本法快速、准确、灵敏、专属性强,适用于同步测定多西他赛和紫杉醇血药浓度及其在中国乳腺癌患者中的药动学研究。