Development of an HPLC–UV–ELSD Method for Quantification of Multiple Components of a Chinese Medicine Made from Radix salvia miltiorrhiza and Panax notoginseng

‘Fufang Danshen tablet’ (FDT), made from Radix salvia miltiorrhiza and Panax notoginseng, is a widely used botanical drug derived from traditional Chinese medicine. Quantification of the active components of Radix salvia miltiorrhiza and Panax notoginseng is very important for regulation of FDT products. In this study HPLC hyphenated with ultraviolet (UV) detection and evaporative light-scattering detection (ELSD) was used for simultaneous determination of nine active components (three salvianolic acids, three tanshinones, and three saponins) of FDT products. Separation was performed on a 250 mm × 4.6 mm i.d., 5.0 μm particle-size, C₁₈ column with linear gradient elution. UV detection at 280 and 254 nm was used for detection of the three salvianolic acids and the three tanshinones, respectively. ELSD was used for detection of the three saponins, which were difficult to analyze by use of UV detection. The linearity of the calibration plots was excellent over the concentration ranges investigated (values of R ² were >0.99 for all the analytes) and recovery measured at three concentrations was between 92.2 and 107.7%. The validated method was successfully used for simultaneous determination of these components in FDT products.     

加速溶剂萃取-气相色谱质谱法测定太子参中酰胺类除草剂的含量

建立了太子参中乙草胺、丁草胺和S-异丙甲草胺的加速溶剂萃取-气相色谱/质谱测定的分析方法。对提取溶剂、萃取温度、净化材料、不同冲洗体积和静态萃取时间、循环次数等实验条件进行了优化。用HP-5MS弹性石英毛细管柱经柱程序升温技术分离,并用质谱检测器检测,内标法计算含量。本方法测定太子参中乙草胺、丁草胺和S-异丙甲草胺的检出限分别为0.16 ng/g、0.18 ng/g和0.05 ng/g,精密度分别为2.6%、3.9%和3.1%,回收率为80.2%-104.1%。所测样品不含上述3种除草剂残留。本方法简便、干扰小、检测效果好,可用于太子参药材中此类除草剂残留的分析。     

One single LC-MS/MS analysis for both phenolic components and tanshinones in Radix Salviae Miltiorrihizae and its medicinal products

A LC-MS/MS method was developed for the separation and simultaneous determination of phenolic components including danshensu, protocatechuic acid, protocatechuic aldehyde and caffeic acid as well as tanshinones including cryptotanshinone, tanshinone I and tanshinone IIA in samples of Radix Salviae Miltiorrhizae and Salviae Miltiorrhizae tablet. Triple quadrupole mass spectrometry was optimized in both positive and negative ion multiple reaction monitoring modes for the simultaneous quantitative analysis of the two different types of active components by using a time-segment program. The method gave recoveries of 85.4-106.4% with relative standard deviations of 2.4-8.0% for the spiked herb samples. The limits of detection were 0.30-0.83 μg/g for the analysis of 1.0 g Radix Salviae Miltiorrhizae or tablet samples.     

保证多指标成分含量稳定的中药材最优化调配方法

提出了一种利用HPLC指纹图谱测定中药材多指标成分含量, 再运用线性、非线性最优化理论调配中药材, 从而保证中药材调配物多指标成分含量稳定的新方法. 方法的核心是所提出的7种最优化调配目标函数及相应的约束条件. 实验验证, 以10批板蓝根药材为基础, 在7种最优化目标下, 控制5个主要HPLC色谱峰面积稳定均一, 结果显示, 峰面积最大相对偏差的绝对值为6.3%.     

气相色谱法测定柴胡注射液中环己醇

建立了气相色谱法测定柴胡注射液中环己醇的定量分析方法，以正己烷为萃取剂，水蒸汽蒸馏浓缩富集提取液，用PEG－20M柱，氢火焰检测器（FID）检测。结果表明，柴胡注射液中环己醇质量浓度在10.0μg/mL-120.0μg/mL范围内呈良好的线性关系（r=0.998），回收率为92 .3%-95.1%，RSD为1.1%－1.2%。     

Separation and determination of two sesquiterpene lactones in Radix inulae and Liuwei Anxian San by microemulsion electrokinetic chromatography

A novel microemulsion electrokinetic chromatography (MEEKC) method for separating and determining two sesquoterpene lactones, alantolactone (AL) and isoalantolactone (IAL), in Radix inulae and Liuwei Anxian San has been developed. The effects of several important factors such as internal organic phases, concentration of microemulsion, concentration of acetonitrile, injection time and running voltage were systematically investigated to determine the optimum conditions. The optimum microemulsion system was composed of n-hexane (0.32% w/w), SDS (1.24% w/w), 1-butanol (2.64% w/w), acetonitrile (10% w/w) and 10 mm sodium tetraborate buffer (85.80% w/w, pH 9.2). The applied voltage was 20 kV. The analytes were detected at 214 nm. Regression equations revealed linear relationships (correlation coefficients 0.9950 for AL and 0.9946 for IAL) between the peak area of each analyte and the concentration. The limits of detection (defined as a signal-to-noise ratio of about 3) were approximately 0.45 microg/mL for AL and 0.56 microg/mL for IAL. The levels of the analytes were successfully determined with recoveries ranging from 98.2 to 104.3%. Furthermore, a simple and effective extraction method, with methanol in an ultrasonic water bath for 60 min, was used for sample preparing. Also, MEEKC was compared with micellar electrokinetic chromatography (MEKC) and shown better separation results.     

A novel method for extraction and separation of total flavones and total astragalosides from <Emphasis Type="Italic">Radix astragali</Emphasis>

In this paper a novel method for extraction and separation of total flavones and total astragalosides from Radix astragali using a resin method is suggested. Ethanol is used as the extractant, the macroporous absorption resin modeled H-107 is used to enrich the astragalus extract, and the macroporous strong base anion resin D-280 is used to separate the total flavones and the total astragalosides. This method allows a good yield of total flavones of 600 mg/100 g Radix astragali, a purity of 94.2%, and a yield of total astragalosides of 28 mg/100 g Radix astragali. Published in Khimiya Prirodnykh Soedinenii, No. 1, pp. 26–29, January–February, 2007.     

黄芩药材中铅的测定及其水煎过程中铅的浸出率研究

测定了黄芩药材中有害元素铅的含量，并研究了黄芩水煎过程中铅的浸出率。     

Simultaneous Determination of Seven Active Compounds in Radix Salviae Miltiorrhizae by Temperature-Controlled Ultrasound-Assisted Extraction and HPLC

A simple and effective high-performance liquid chromatographic (HPLC) method has been developed for simultaneous quantification of three phenolic acids (3,4-dihydroxyphenyllactic acid (Chinese name danshensu), protocatechuic aldehyde, and salvianolic acid B) and four diterpenes (dihydrotanshinone I, cryptotanshinone, tanshinone I, and tanshinone II_A) in radix salviae miltiorrhizae. Chromatography was performed on a 250 mm × 4.6 mm i.d., 5-μm particle size, C₁₈ column. The mobile phase was a linear gradient prepared from 0.1% (v/v) aqueous formic acid and acetonitrile at a flow-rate of 1.0 mL min⁻¹. All the target components were well separated with high resolution and without interference. Good linearity (R ² > 0.999) was observed over the concentration ranges investigated, and intra-day and inter-day precision were high. Temperature-controlled ultrasound-assisted extraction was used to prevent hydrolysis of thermally unstable components during the sample-extraction procedure, and the extraction conditions were carefully optimized. Recovery of the seven components was from 98.45 to 100.63% and relative standard deviations were always <1.5%. The validated method was successfully used for simultaneous quantification of the three phenolic acids and the four diterpenes in radix salviae miltiorrhizae of different geographic origins.     

A rapid method for simultaneous determination of 14 phenolic compounds in Radix Puerariae using microwave-assisted extraction and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry

A microwave-assisted extraction (MAE) and ultra high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry (UHPLC-DAD-TOF-MS) method was developed for simultaneous determination of 14 phenolic compounds in the root of Pueraria lobata (Wild.) Ohwi and Pueraria thomsonii Benth. Operational conditions of MAE were optimized by central composite design (CCD). The optimized result was 65% ethanol as extraction solvent, 17 mL of extraction volume, 100 °C of extraction temperature and 2 min of hold time. A Zorbax SB C₁₈ (50 mm × 4.6 mm I.D., 1.8 μm) and gradient elution were used during the analysis. The chromatographic peaks of 14 investigated compounds in samples were successfully identified by comparing their retention time, UV spectra and TOF mass data with the reference substances. All calibration curves showed good linearity (r > 0.9997) within the test ranges. The intra-day and inter-day variations were less than 1.77% and 2.88%, respectively. The developed method was successfully applied to determine the investigated compounds in 10 samples of Radix Puerariae Lobatae and Radix Puerariae Thomsonii, respectively. The result indicated that MAE and UHPLC-DAD-TOF-MS system might provide a rapid method for the quality control of Radix Puerariae.