Comparative analysis of nucleosides,nucleobases, and amino acids in different parts of Angelicae Sinensis Radix by ultra high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry

A rapid and sensitive ultra high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry method was established and employed to determine 21 nucleosides, nucleobases, and amino acids in 60 samples from different parts of Angelicae Sinensis Radix. The established methods were validated by good linearity (r² > 0.9937), limits of detection (0.12–77.75 ng/mL), limits of quantitation (0.31–272.13 ng/mL), intra‐ and interday precisions (RSD ≤ 4.84%, RSD ≤ 6.26%), stability (RSD ≤ 5.92%), repeatability (RSD ≤ 7.14%), recovery (91.4–103.4%), and matrix effects (0.92–1.03). Chemical comparative analysis revealed that the content of total analytes in four parts of Angelicae Sinensis Radix were different, and exhibited the order: Head (14.89 mg/g) > Body (10.15 mg/g) > All (8.22 mg/g) > Tail (6.23 mg/g). Principal component analysis showed that the samples could be classified into four groups in accord with four different parts of Angelicae Sinensis Radix. The results could provide a scientific basis and reference for the quality control of Angelicae Sinensis Radix, and may be conducive to further research on the pharmacological activities of Angelicae Sinensis Radix.     

西洋参药材质量评价研究进展

对影响西洋参药材质量的主要因素如产地、采收期和干燥方法等方面进行了综述。认为目前西洋参药材质量评价研究中存在的关键问题是评价指标过于片面单一，未来质量评价研究应考虑引入质量整体评价技术，注重中医药理论，开展有效成分、药理作用与中药功效的整合研究。药材质量研究应指导药用植物的产地筛选、生长调控，服务于药材市场的规范，以促进中药现代化进程的发展。     

A purge and trap technique to capture volatile compounds combined with comprehensive two‐dimensional gas chromatography/time‐of‐flight mass spectrometry to investigate the effect of sulfur‐fumigation on Radix Angelicae Dahuricae

Sulfur‐fumigation is known to reduce volatile compounds that are the main active components in herbs used in herbal medicine. We investigated changes in chemical composition between sun‐dried and sulfur‐fumigated Radix Angelicae Dahuricae using a purge and trap technique to capture volatile compounds, and two‐dimensional gas chromatography/time‐of‐flight mass spectrometry for identification. Using sun‐dried Radix Angelicae Dahuricae samples as a reference, the results showed that 73 volatile compounds, including 12 sulfide compounds, were found to be present only in sulfur‐fumigated samples. Furthermore, 32 volatile compounds that were found in sun‐dried Radix Angelicae Dahuricae samples disappeared after sulfur‐fumigation. The proposed method can be applied to accurately discriminate sulfur‐fumigated Radix Angelicae Dahuricae from different commercial sources. Copyright © 2014 John Wiley & Sons, Ltd.     

超高效液相色谱-串联质谱法快速检测洛党参中乙基多杀菌素残留

建立了超高效液相色谱-串联质谱(UHPLC-MS/MS)快速检测洛党参中乙基多杀菌素-J(XDE-175-J)和乙基多杀菌素-L(XDE-175-L)残留的方法。样品经乙腈提取,乙二胺-N-丙基硅烷和石墨化炭黑净化后,外标法定量。结果表明,XDE-175-J和XDE-175-L的峰面积与其质量浓度分别在0.075~75μg/L和0.025~25μg/L范围内,呈良好线性关系(r20.990);在0.375、3.75、75μg/kg加标水平下,XDE-175-J的平均回收率为88.4%~113.5%,相对标准偏差(RSD,n=5)为2.0%~4.2%;在0.125、1.25、25μg/kg加标水平下,XDE-175-L的平均回收率为84.4%~99.5%,RSD(n=5)为2.5%~4.9%。乙基多杀菌素的定量下限(LOQ)为0.375μg/kg(XDE-175-J)和0.125μg/kg(XDE-175-L)。该方法简便、快捷,灵敏度高,回收率和重复性良好,能满足农药残留检测技术要求,可用于大量洛党参样品中乙基多杀菌素的残留分析。     

葛根-丹参组方对动脉粥样硬化鹌鹑血管内皮组织 的影响

目的：考察葛根-丹参(1:1)组方对动脉粥样硬化鹌鹑血管内皮组织的影响. 方法: 连续喂养高脂饲料造模同时给药, 观察葛根-丹参对动脉粥样硬化鹌鹑主动脉组织脂质斑块、内皮及内膜组织的影响. 结果: 葛根-丹参组方配伍后, 无论乙酸乙酯部位还是水提物, 均能较好的减少泡沫细胞及脂质的沉积, 改善内皮细胞的肿胀、变性, 抑制血管内膜的增厚及巨噬细胞的数量, 尤以乙酸乙酯部位作用明显. 结论: 葛根丹参组方配伍具有一定的减少主动脉组织脂质斑块形成, 保护血管内皮细胞作用, 有效地阻止AS的形成与发展.     

Simultaneous determination of 15 marker constituents in various radix Astragali preparations by solid-phase extraction and high-performance liquid chromatography

An improved quality control method was developed to simultaneously determine 15 major constituents (eight flavonoids and seven saponins) in various radix Astragali preparations, using SPE for pretreatment of samples, HPLC with diode-array and evaporative light scattering detectors (DAD-ELSD) for quantification in one run, and HPLC-ESI-TOF/MS for definite identification of compounds in preparations. Optimum separations were obtained with a ZORBAX C(18) column, using a gradient elution with 0.3% aqueous formic acid and ACN. This established method was fully validated with respect to linearity, precision, repeatability, and accuracy, and was successfully applied to quantify the 15 compounds in 19 commercial samples, including 3 dosage forms, i. e., oral solution, injection, concentrated granule, and its processed products of radix Astragali. The results demonstrated that many factors might result in significant differences in quality of the final preparations, including crude drugs, pretreatment processes, manufacturing procedure, storage conditions, etc. Then the developed method provided a reasonable and powerful manner to ensure the efficacy, safety, and batch-to-batch uniformity of radix Astragali products by standardizing each procedure, and thus should be proposed as quality control for the clinical use and modernization of herbal preparations.     

Quick analysis of baicalin in Scutellariae Radix by enzyme-linked immunosorbent assay using a monoclonal antibody

To establish an immunoassay for baicalin (BA), a hybridoma cell line (9D6) secreting a monoclonal antibody (MAb) against BA was prepared by cell fusion with splenocytes derived from a mouse immunized with BA-bovine serum albumin (BSA) conjugate and a myeloma cell line, SP2/0-Ag14. MAb 9D6 shows specific reactivity against BA and its aglycone, baicalein, but not against other natural products. We developed an enzyme-linked immunosorbent assay (ELISA) using MAb 9D6 in a competitive manner, ranging from 200 ng/mL to 2 μg/mL. After validating the developed ELISA on the basis of intra- and inter-assays and a recovery experiment, it was found that the ELISA was not only simple, but also sufficiently reliable and accurate for quality control of Scutellariae Radix. It allowed determination of BA in complex and mixed materials, such as Kampo medicines.     

Simultaneous determination of harpagoside and cinnamic acid in rat plasma by liquid chromatography electrospray ionization mass spectrometry and its application to pharmacokinetic studies

A simple and sensitive method was developed for the simultaneous quantification of harpagoside and cinnamic acid in rat plasma using high-performance liquid chromatography system coupled to a negative ion electrospray mass spectrometric analysis. The plasma sample preparation was a simple deproteinization by the addition of two volumes of acetonitrile. The analytes were separated on an Intersil C8-3 column (2.1 mm i.d.x250 mm, 5 microm) with acetonitrile-5 mm ammonium formate aqueous solution (60:40, v/v) as mobile phase at a flow-rate of 0.2 mL/min. Detection was performed on a quadrupole mass spectrometer equipped with electrospray ionization (ESI) source operated under selected ion monitoring (SIM) mode. [M+HCOO]- at m/z 539 for harpagoside, [M-H]- at m/z 147 for cinnamic acid and [M-H]- at m/z 137 for salylic acid (internal standard) were selected as detecting ions, respectively. The method was validated over the concentration range 7-250 ng/mL for harpagoside and 5-500 ng/mL for cinnamic acid. The lower limits of quantitation for harpagoside and cinnamic acid were 7 and 5 ng/mL, respectively. The intra- and inter-day precisions (RSD%) were within 9.5% and the assay accuracies (RE%) ranged from -5.3 to 3.0% for both analytes. Their average recoveries were greater than 86%. Both analytes were proved to be stable during all sample storage, preparation and analysis procedures. The method was successfully applied to the pharmacokinetic study of harpagoside and cinnamic acid following oral administration of Radix Scrophulariae extract to rats.     

Analysis of 14 terpenoids and sterols and variety discrimination of Codonopsis Radix using ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry

Recently, we confirmed that the 95% ethanol-extracted fraction of Codonopsis Radix, which contains several triterpenoids and sterols, possesses pharmacological activities. However, due to the low content and diverse types of triterpenoids and sterols, their similar structures, lack of ultraviolet absorption, and difficulty in obtaining controls, few studies have so far assessed their contents in Codonopsis Radix. We accordingly constructed an ultra-high-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry technique for the simultaneous quantitative determination of 14 terpenoids and sterols. Separation was performed on the Waters Acquity UPLC HSS T3 C₁₈ column (100 × 2.1 mm, 1.8 μm) with 0.1% formic acid (A) and 0.1% formic acid in methanol (B) as mobile phase under gradient elution. The determination coefficients for each of the matrix calibration curves were ≥0.9925. The average recovery ranged from 81.25% to 118.05%, with relative standard deviations of <4%. The contents of 14 components in 23 batches were quantified and further analyzed through chemometrics. Linear discriminant analysis can distinguish sample varieties. The quantitative analysis method can accurately determine the contents of 14 components and thereby provide the chemical basis for the quality control of Codonopsis Radix. It also could be a valuable approach for the classification of different Codonopsis Radix varieties.     

Qualitative identification and quantitative comparison of Physochlainae Radix from different regions based on chemometric methods

Physochlainae Radix (PR) is an essential herbal medicine that has been generally applied for treating cough and asthma. In this study, a comprehensive strategy for quality evaluation of PR from different origins was established by integrating qualitative identification, quantitative analysis, and chemometric methods. A total of 58 chemical components were identified by ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS/MS), and a sensitive and rapid UHPLC-QqQ-MS/MS method was established for the simultaneous determination of 12 compounds. In addition, multivariate statistical analysis was applied for discriminant analysis to compare the differences among 30 batches of PR samples. The results showed that the 30 batches of PR collected from four provinces could be clustered into three categories, in which scoparone, protocatechuic acid, tropic acid, and scopolin were important components to distinguish the primary and non-primary producing areas, as well as superior and inferior products of PR. Chemometric results were consistent and validated each other, and systematically explained the intrinsic quality characteristics of PR. This study first demonstrated that LC-MS combined with multivariate statistical analysis, provided a comprehensive and effective means for quality evaluation of PR.