目的：建立高效液相色谱分离检测脑蛋白水解物注射液各种氨基酸的测定方法。方法：氨基酸含量测定采用异硫氰酸苯酯柱前衍生化；色谱柱为C18柱（VP-DOS；150 mm×46 mm）；流动相：A为0.1 mol/L醋酸钠溶液（冰醋酸调节pH 6.5）-乙腈（93∶7），B为水-乙腈（1∶4）；检测波长：254 nm；流速：1 mL/min；柱温：40℃。结果：16种氨基酸在32 min内完全分离，各种氨基酸之间没有出现干扰现象，且多次重复测定结果误差很小。结论：建立的高效液相色谱法准确性高、重现性好，可以有效地控制脑蛋白水解物注射液的质量。
Objective To determine the best threshold value of hemoglobin A2 (HbA2) for diagnosis of β-thalassemia (β-thal) carriers by using high performance liquid chromatography (HPLC), and to improve the application value of HbA2 as a diagnostic index for β-thal carriers to reduce the rates of missed diagnosis and misdiagnosis. Methods Using reverse dot blot (RDB) as a gold standard method, HbA2 results of 1 007 β-thal carriers and 606 normal controls in the past two years determined by HPLC were divided into true positive, false positive, true negative, and false negative based on the different threshold values of HbA2 results. Then, the evaluation indexes such as sensitivity, specificity, positive and negative likelihood ratio, and Youden’s index were evaluated. Next, the receiver operator characteristic (ROC) curve was drawn to determine the best threshold value of HbA2 for diagnosis of β-thal carriers by HPLC. Results If ≥4.0% was taken as the threshold value of HbA2 for diagnosis of β-thal carriers by HPLC, the evaluation indexes values were shown as follows: sensitivity 99.21%, specificity 99.34%, positive likelihood ratio 150.30, negative likelihood ratio 0.008, and Youden’s index 0.99. The Youden’s index was better than the other threshold values, and the corresponding tangent point was the peak point of the ROC curve. Conclusion When ≥4.0% serves as the best threshold value of HbA2 for diagnosis of β-thal carriers using HPLC, integrated evaluation performance of the corresponding sensitivity and specificity is the most ideal, and the authenticity of the diagnostic test is the best.
目的 建立测定胎盘灌流液中格列苯脲浓度的高效液相色谱（HPLC）检测方法。方法 采用的色谱柱为Symmetry Shield RP C18（150 mm×4.6 mm，5 μm），柱温40℃，流动相为NaH2PO4缓冲盐（25 mmol/L，pH值5.2）︰乙腈=1︰1；内标为格列齐特，流速1.0 mL/min，检测波长228 nm，采用液-液萃取预处理方法测定胎盘灌流液中格列苯脲的浓度。 结果 格列苯脲浓度线性范围为2.0～25.0 μg/mL，线性方程为：y=0.226x+0.002，r=0.999 9 （n=6），日内相对标准偏差（RSD）＜3.1%，日间RSD＜9.5%，方法学回收率为95.32%～103.35%。 结论 HPLC检测方法灵敏、简便，可用于胎盘灌流液中格列苯脲浓度的检测。
目的 建立柱前衍生化反相高效液相色谱-荧光检测法测定血浆中同型半胱氨酸（Hcy）浓度的方法。 方法 以tris-(2-carboxylethyl)-phosphine hydrochloride （TCEP）为还原剂，以7-fluorbenzo-2-oxa-1,3-diazole-4-sulfonate （SBD-F）为衍生剂，色谱柱为Xterra RP18柱，柱温35℃，流动相为甲醇︰磷酸二氢钠缓冲液（pH值3.0）=3︰97，激发波长380 nm，发射波长510 nm，外标法定量。 结果 Hcy浓度在1.95～125 ?mol/L范围内线性关系良好（r=0.999 8）。日内和日间相对标准偏差均＜7%，方法回收率为103.2%～111.9%。 结论 此方法准确、灵敏、快速，是一种适合于实验室研究和临床检测血浆中Hcy浓度的方法。
目的 采用高效液相色谱法测定受试者口服埃索美拉唑肠溶胶囊与埃索美拉唑镁肠溶片后血药浓度，评价埃索美拉唑肠溶胶囊的生物等效性。 方法 2009年9月－10月，36例健康男性受试者单次交叉口服埃索美拉唑肠溶胶囊（试验制剂）和埃索美拉唑镁肠溶片（参比制剂），测定给药后不同时间点血浆中埃索美拉唑经时血药浓度，采用DAS 2.0软件进行药物代谢动力学参数计算和生物等效性评价。 结果 受试者单次口服试验制剂与参比制剂后，达峰时间分别为（2.19 ± 0.96）、（2.43 ± 0.92） h，峰浓度分别为（1 748.86 ± 615.81）、（1 442.92 ± 476.41） μg/L，药时曲线下面积（AUC）0-t分别为（3 927.14 ± 1 839.10）、（3 878.79 ± 1 734.84） μg/L·h，AUC0-∞分别为（3 998.36 ± 1 866.22）、（3 918.31 ± 1 773.44） μg/L·h。试验制剂与参比制剂的生物等效性为94.0%，其90%CI为（82.3%，107.2%）。 结论 埃索美拉唑肠溶胶囊与埃索美拉唑镁肠溶片生物等效。
【摘要】 目的 考察用乌头碱水解物、新乌头碱水解物、次乌头碱水解物的含量作为附片水煎剂及附片质量控制指标的可行性。 方法 采用高效液相色谱分析法，色谱柱为Shimpack CLC-ODS，以甲醇-乙酸铵溶液（0.2 mol/L）（200︰210）为流动相，流速1.0 mL/min，检测波长为241 nm，对乌头碱新，乌头碱、次乌头碱水解物的含量测定进行研究。 结果 3种水解物检测方法测得水解物在进样范围有良好线性关系，重现性好，稳定性好，生药提取时间宜选定为40 min，不同批号的附片中3种水解物含量差别较大。 结论 此方法简便，使用新乌头碱、乌头碱、次乌头碱的水解物作为含附片制剂的质量控制指标是可行的。【Abstract】 Objective To examine the feasibility of regarding the hydrolyzate from aconitine, new aconitine, and hypaconitine to be the decoction of radix aconite and the quality control index. Methods High performance liquid chromatographic method (HPLC) method was adopted. Shimpack CLC-ODS column with UV detection at 241 nm was used. The mobile phase consisted of (200∶210), and the flow rate was 1.0 mL/min. Results The content of the hydrolysis objects in the water decoction and toxicity were correlated with each other, and the contents of hydrolysis were significantly different among different groups of radix aconiti. Conclusion This method is simple, accurate and effective for content research of hydrolyzate from aconitine-type alkaloids in Monkshoo decoction of Radix Aconiti, and it is feasible to take it as the quality control index.
目的:建立以高效液相色谱法测定枣仁安神颗粒中丹酚酸B含量的方法。方法:色谱柱为phenomenex Lunar C-18(150mm×4.6mm id,5μm),流动相为甲醇-乙腈-甲酸-水(30∶10∶1∶59),流速为1.0mL·min-1,检测波长为286nm。结果:丹酚酸B进样量在0.1664～1.6640μg范围内与峰面积分值呈良好的线性关系(r=0.9999);平均加样回收率为99.95%,RSD=0.09%(n=9)。结论:本方法操作简便,准确,可用于该制剂中丹酚酸B的含量测定。
Objective To study the distribution and concentration of meropenem in rabbit bile. Methods The rabbits were cannulated with a silicone tube in the common bile duct and the blank bile was collected. The rabbits were then administered intravenously with meropenem. Multiple bile samples (1.5 ml) were collected at different phases after the administrations. According to requirement of the high performance liquid chromatography (HPLC), the specificity test was undertook. The blank bile was then mixed with meropenem and mobile phase, respectively, in order to obtain a series of bile samples at different concentrations ranging from 0.5 to 500 μg/ml. The samples were analyzed with HPLC and the chromatographic peak area of meropenem contents were quantitated through external reference method. The linear regression equation was used to analyzed the relationship between the drug concentrations and the chromatographic peak areas. The bile samples that were collected after drug administrations were pretreated and the chromatographic peak areas were assayed by the liquid chromatograph. The bile concentrations were then calculated according to the regression equation, and the concentration-time distribution of meropenem in the bile was obtained ultimately. Results The specificity test indicated the bile dopant peak and the meropenem chromatographic peak were well-separated under chromatographic condition of the mobile phase. The standard curve regression equation was S=2 209.10C-1 251.34, r=0.999 9, and minimum quantitation limit of meropenem was 0.5 μg/ml. After a single i.v. administration of 75 mg/kg of meropenem in each rabbit, drug concentrations reached (38.36±14.17) μg/ml immediately in bile， which significantly exceeded the minimum inhibitory concentrations (MIC90) for most gram negatives, which range from 0.031 to 2 μg/ml. The bile concentration of meropenem decreased quickly over time, and meropenem was eliminated completely in rabbit bile 3 hours after intravenous injection. Conclusion Meropenem could achieve adequate bile concentration for the treatment of biliary tract infection due to susceptible bacteria. However, because of its rapid biliary elimination, meropenem should be used in shorter interval time.
ObjectiveTo determinate the concentration of isonicotinyl hydrazide (INH) in the hydrothorax and thereby increase the drug concentration of the hydrothorax to enhance the anti-tuberculosis efficacy through experiments. MethodsBetween February and June 2009, we used high performance liquid chromatography (HPLC) to determinate the concentration of INH in the hydrothorax of tuberculosis (TB) patients. The separation of sample was performed on Agilent ZORBAX SB-C18 (5 μm, 4.6 mm×2.5 mm) column with a fixed sample injection volume of 20 μL. The mobile phase was methanol-water (20︰80) at a flow rate of 1 mL/min. The ultraviolet detective wavelength was set at 254 nm; The column temperature was room temperature. ResultsIn the range of 0.25-10.00 μg/mL (r=0.998 9), moxi-floxacin showed a good linear relation in HPLC. The recoveries of moxi-floxacin at three concentrations (0.5, 5.0, and 10.0 μg/mL) were 97.95%, 100.64%, and 102.84%, respectively, with intra-day relative standard deviation (RSD) at 3.49%, 1.45%, and 2.03%, respectively and intra-day RSD at 3.85%, 5.68%, 4.15%, respectively. ConclusionThe measuring method adopted in the experiment can accurately determinate the concentration of INH in the hydrothorax with high sensitivity and exceptional reproducibility.
ObjectiveTo establish a method for content determination of ferulic acid in Piwang massage lotion. MethodsHigh performance liquid chromatography was used. The analysis was carried on Diamonsil C18 (150 mm×4.6 mm, 5 μm, Dimma Science and Technology Co. Ltd.) column with mobile phase consisted of methanol-2% acetic acid (32︰68). The column temperature was 30℃. The detection wavelength was 320 nm. ResultsThe responses of ferulic acid liner in range of 4.55-91.00 μg/mL (R2=0.999 9). The average recovery of ferulic acid was 98.78% (relative standard deviation is 2.668%). ConclusionThe method is simple, rapid and good repeatability. It can be used for quality control of Piwang massage lotion