安胃片质量标准研究

对安胃片进行质量标准研究。采用薄层色谱法对安胃片中延胡索进行鉴别,并且采用高效液相色谱法对其中的延胡索乙素进行含量测定。色谱柱为Kromasil C18色谱柱（4.6mm×150mm,5μm）,流动相为甲醇-水（三乙胺调pH值为8.68,V：V=65：35）,检测波长为280nm,流速1.0mL/min,柱温为室温。结果显示在薄层色谱中均可检出延胡索特征斑点;延胡索乙素在0.18—1.47μg范围内线性关系良好,r=0.9999,平均加样回收率为97.49%（RSD=1.64%,n=6）。本方法简便可行、重复性好,能有效控制该制剂的质量。     

胃复宁片的质量标准

建立胃复宁片的质量标准。采用薄层色谱法对胃复宁片中的延胡索和白术进行鉴别;采用HPLC测定胃复宁片中延胡索乙素的含量。薄层色谱斑点清晰,阴性对照无干扰;延胡索乙素含量在0.22—1.10μg范围内与峰面积呈良好线性关系(r=0.9992);平均回收率为97.1%,RSD=2.09%(n=6)。方法简单、准确、重现性好,可有效地控制胃复宁片的质量。     

Chiral high-performance liquid chromatographic separation of the enantiomers of tetrahydropalmatine and tetrahydroberberine, isolated from Corydalis yanhusuo

HPLC methods have been developed for chiral resolution of the enantiomers of dl-tetrahydropalmatine (THP) and dl-tetrahydroberberine (THB), two active constituents of Corydalis yanhusuo W.T. Wang. On the analytical scale, good baseline separation of the enantiomers was achieved using cellulose tris(3,5-dimethylphenylcarbamate) chiral stationary phases in both normal-phase and polar organic modes. Validation of the analytical methods, including linearity, limits of detection, recovery, and precision, and semipreparative resolution of dl-THP and dl-THB, were achieved with methanol as mobile phase, without any basic additives, in polar organic mode using cellulose tris(3,5-dimethylphenylcarbamate) chiral stationary phases. On the semipreparative scale, small quantities of the individual enantiomers of THP and THB were isolated for study of the chiroptical properties of the individual enantiomers.     

Affinity capillary electrophoresis coupling with partial filling technique and field‐amplified sample injection for enantioseparation and determination of DL‐tetrahydropalmatine

A novel, simple and sensitive method for the enantioseparation and determination of DL ‐tetrahydropalmatine (DL ‐THP) was developed using ACE in combination with partial filling technique and field‐amplified sample injection. A chiral selector, i.e. BSA, was used for the enantioseparation of DL ‐THP in ACE. Effects of BSA concentration, pH and separation voltage on the effectiveness of the enantiomer separation were evaluated. In an optimal condition, D ‐ and L ‐THP were completely enantio‐separated in less than 9 min by partially filling an electrophoretic capillary with 50 μmol/L BSA (50 mbar, 100 s) and carrying out an electrophoresis with 20 mmol/L phosphate buffer (pH 7.4) at 15 kV. The sensitivity was further improved by making use of field‐amplified sample injection to lower the LOD (defined as S/N=3) down to 6 ng/mL. Real samples were also tested and promising results for the determination of DL ‐THP enantiomers were obtained.     

Application of nonaqueous capillary electrophoresis for quantitative analysis of quinolizidine alkaloids in Chinese herbs

An easy, rapid method for simultaneous determination of tetrandrine (TET), fangchinoline (FAN), sinomenine (SIN) and tetrahydropalmatine (TEP) in Chinese herbs was developed by nonaqueous capillary electrophoresis without pretreatment for the first time. Optimum separation was achieved with a fused-silica capillary column ( i.d.) and a running buffer containing 50 mM ammonium acetate, 1.0% acetic acid and 20% acetonitrile in methanol medium. The applied voltage was 20.0 kV. The analytes were detected by UV at 214 nm. The effects of concentration of ammonium acetate, acetic acid and organic modifier on electrophoretic behavior of the analytes were studied. Regression equations revealed linear relationships (correlation coefficients: 0.9991-0.9999) between the peak area of each analyte and the concentration. The levels of analytes in Stephania tetrandra S. Moore, Rhizoma corydalis and Sinomenium acutum Rehd. et. Wils were easily determined with recoveries ranging from 96 to 107%.     

Simultaneous determination of tetrahydropalmatine and tetrahydroberberine in rat urine using dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography

Dispersive liquid-liquid microextraction (DLLME) coupled with high-performance liquid chromatography (HPLC)-UV detection was applied in rat urine for the extraction and determination of tetrahydropalmatine (THP) and tetrahydroberberine (THB), both active components in Rhizoma corydalis. Various parameters affecting the extraction efficiency, such as the type and volume of extraction and dispersive solvent, pH, etc. were evaluated. Under the optimal conditions (extraction solvent: 37 μL of chloroform, dispersive solvent: 100 μL of methanol, alkaline with 100 μL of 1 mol/L NaOH, and without salt addition), the enrichment factors of THP and THB were more than 30. The extraction recoveries were 69.8-75.8% and 72.7-77.6% for THP and THB in rat urine, respectively. Both THP and THB showed good linearity in the range of 0.025-2.5 μg/mL, and the limit of quantification was 0.025 μg/mL (S/N=10, n=6). The intra-day and inter-day precision of THP and THB were <12.6%. The relative recoveries ranged from 95.5 to 107.4% and 96.8 to 100.9% for THP and THB in rat urine, respectively. The method has been successfully applied to rat urine samples. The results demonstrated that DLLME is a very simple, rapid and efficient method for the extraction and preconcentration of THP and THB from urine samples.     

延胡索乙素在兔体内脏的分布滞留研究

本文为了对药物在脏器中的分布滞瘤问题进行研究，首先给家兔口服元胡水煎液，２４ｈ之后，再采用高灵敏的胶束荧光法，对主要脏器中所含延胡索乙素进行测定，所测心肝、肾中的含量分别为０．０７９８μｇ／ｇ，１．４０μｇ／ｇ，５．１４μｇ／ｇ，效果良好。     

High-Performance Liquid Chromatography with Electrospray Mass Spectrometry for Rapid and Sensitive Determination of Sanguinarine and Chelerythrine in Exogenously Contaminated Honey

A simple, rapid, and sensitive high-performance liquid chromatographic (HPLC) method coupled with electrospray mass spectrometry (ESI-MS) has been used to determine sanguinarine and chelerythrine in exogenously contaminated honey. Sample extracts were separated on a C₈ reversed-phase HPLC column with acetonitrile–acetate buffer (40:60) as mobile phase. After ESI the abundance of protonated molecules was recorded by selected-ion recording (SIR) of m/z 332.5, 348.5, and 356.5 for sanguinarine, chelerythrine, and the internal standard, tetrahydropalmatine, respectively. The internal standard technique was used to construct calibration plots for quantitation of sanguinarine and chelerythrine; the linear ranges were 5.25–1050 and 3.75–750 ng mL^–1, respectively, with correlation coefficients of 0.9993 and 0.9989, respectively. The limits of detection for sanguinarine and chelerythrine were 1.60 and 1.11 ng mL^–1, respectively.     

Simultaneous quantification of l‐tetrahydropalmatine and its urine metabolites by ultra high performance liquid chromatography with tandem mass spectrometry

l ‐tetrahydropalmatine (l ‐THP) is a tetrahydroprotoberberine isoquinoline alkaloid that has been used as an analgesic agent in China for more than 40 years. Recent studies indicated its potential application in the treatment of drug addiction. In this study, a sensitive and rapid method using ultra high performance liquid chromatography with MS/MS was developed and validated for simultaneous quantitation of l ‐THP and its desmethyl metabolites. Enzymatic hydrolysis was integrated into sample preparation to enable the quantitative determination of both free and conjugated metabolites. Chromatographic separation was achieved on an Agilent Poroshell 120 EC‐C₁₈ column. Detection was performed by MS in the positive ion ESI mode. The calibration curves of the analytes were linear (r² > 0.9936) over the concentration range of 1–1000 ng/mL with the lower limit of quantification at 1 ng/mL. The precision for both intra‐ and interday determinations was <8.97%, and the accuracy ranged from ?8.74 to 8.65%. The recovery for all the analytes was >70% without significant matrix effect. The method has been successfully applied to the urinary excretion study of l ‐THP in rats. The conjugates were found to be the major urine metabolites of the drug.