Quantification of tripdiolide in human whole blood by liquid chromatography coupled with atmospheric pressure chemical ionization tandem mass spectrometry

A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) assay was developed and validated to determine tripdiolide in human whole blood using dexamethasone acetate as an internal standard (I.S.). Liquid-liquid extraction with ethyl acetate was used to isolate them from the biological matrix. Detection was performed on a mass spectrometer coupled with a negative atmospheric pressure chemical ionization (APCI) in the multiple-reaction monitoring (MRM) mode. The calibration curve was linear (r² = 0.9973) in the concentration range of 0.5-100.0 ng/mL in human whole blood with a lower limit of quantification of 0.5 ng/mL. Intra-day and inter-day relative standard deviations (R.S.D.s) were less than 7.0 and 10.1%, respectively. Extraction recoveries of tripdiolide ranged from 80.5 to 90.1%. This assay can be used to determine trace tripdiolide in human whole blood.     

Simultaneous Determination of Triptolide and Tripdiolide in Extract of Tripterygium wilfordii Hook. f. by LC–APCI-MS

A novel method using liquid chromatography coupled with atmospheric pressure chemical ionization mass spectrometry in the negative selected ion monitoring mode has been developed and validated for rapid simultaneous determination of triptolide and tripdiolide in the extract of Tripterygium wilfordii Hook. f. The molecular ions m/z [M–H]⁻ 359 and 375 were selected for the quantification in selected ion monitoring mode for triptolide and tripdiolide. Standard calibration curve was linear over the concentration range of 0.12–24 and 0.15–30 μg mL⁻¹ for triptolide and tripdiolide. The relative standard deviations of intra- and inter-day were in the range of 4.7–9.9 and 8.9–12.6%. The average recoveries were between 96.4 and 104.6%. The limits of quantitation were 2.0 × 10⁻³ and 2.5 × 10⁻³ μg mL⁻¹ for triptolide and tripdiolide.     

雷酚内酯的极谱行为研究

研究雷酚内酯在多种体系中的极谱行为。实验表明 ,在不同体系和不同pH范围 ,雷酚内酯产生两个还原波 ,其峰电流和峰电位受酸度影响的情况不同 ,而在B -R缓冲体系条件下 ,峰电流明显受溶液酸度的影响 ,但峰电位基本上保持不变 ;在pH =6.1的B -R缓冲溶液中 ,雷酚内酯的两峰电位分别为 -0 .1 1V和-1 .1V(vs.SCE) ,两峰的峰电流与雷酚内酯的浓度在 3 .1 2 9× 1 0 - 6 mol·L- 1～ 3 .1 2 9× 1 0 - 5mol·L- 1的范围内成良好的线性关系 ,线性回归方程分别为iP′(nA·s- 1) =3 1 1 6+2 .447× 1 0 7c(mol·L- 1) (r =0 .9994)和iP′(nA·s- 1) =1 3 5 2 +4 .83 3× 1 0 7c(mol·L- 1) (r =0 .9993 ) ;循环伏安研究表明 ,两波都是不可逆吸附波     

Two new terpenes from Tripterygium wilfordii

<正>Two new terpenes,wilfornine H(1) and triptobenzene Q(2) were isolated from Tripterygium wilfordii,as well as 11 terpenes. Their structures were elucidated by spectroscopic methods.Compounds 5,9-13 showed significant immunosuppressive activities.     

Two New Abietane Diterpenoids from the Roots of Tripterygium wilfordii Hook. f.

Two new abietane‐type diterpenoids, named triptobenzene R ( 1 ) and triptobenzene S ( 2 ), together with three known abietane‐type diterpenoids, triptophenolide ( 3 ), triptonodiol ( 4 ), and triptonoterpene methyl ether ( 5 ), were isolated from the roots of Tripterygium wilfordii Hook . f. Their structures and relative configurations were established by detailed spectral studies, including 1D‐ and 2D‐NMR (HSQC, HMBC, and NOESY), and HR‐ESI‐TOF‐MS, and by comparison with published data. Their absolute configurations were assigned by the CD technique, applied for the first time to abietane diterpenes from Tripterygium wilfordii. Compound 2 is the first abietane‐type norditerpenoid isolated from the genus Tripterygium.     

应用UPLC-Q-TOF-MS/MS技术分析生物诱导后雷公藤悬浮细胞化学成分

利用超高效液相色谱-四级杆-飞行时间质谱连用技术(UPLC-Q-TOF-MS/MS)和化学计量学结合的方法,分析生物诱导后雷公藤悬浮细胞体系中化学成分的变化.分析样品相似度、层次聚类分析(HCA)和主成分分析(PCA),建立554个共有峰的质谱指纹图谱.结果表明:通过对雷公藤离体培养体系中的质谱指纹图谱分析,鉴定了104种化学成分;生物诱导后雷公藤悬浮细胞和培养液中部分化学成分质量分比显著升高,雷公藤内酯乙、山海棠酸衍生物1、TH1、雷公藤甲素衍生物2、雷酚内酯、雷公藤甲素衍生物1、雷酚新内酯质量比分别提高了7.41,6.50,6.07,5.85,3.63,1.59,1.13倍;质荷比(m/z)为224,278,678,701的未鉴定成分质量比分别提高了2.88,1.57,4.04,6.27倍.     

A Novel Diterpenoid,11—O—β—D—glucopyranosylneo—triptophenolide, from Tripterygium wilfordii Hook.f

A new diterpenoid was isolated from the ethanolic extract of the dried root bark of Tripterygium wilfordii Hook.f. It is the first example of abietane diterpenoid glycoside isolated from Tripterygium wilfordii Hook.f.Its structure was identified to be 11-0-β-D-glucopyranosyl-neotriptophenolide based on spectral methods.     

Separation and simultaneous determination of seven bioactive components in Tripterygium wilfordii Hook. F. and Tripterygium preparations by micellar electrokinetic capillary chromatography

A simple, comprehensive, and highly selective MEKC method has been developed for simultaneous analysis of seven bioactive components (triptolide, wilfortrine, wilfordine, wilforgine, wilforine, triptophenolide, and triptonide) in the root extracts of Tripterygium wilfordii Hook. F. (TWHF) and Tripterygium preparations (TPs). Optimal BGE consisted of 10 mM sodium tetraborate, 30 mM SDS, and 30% v/v methanol. The separation voltage was 20 kV and the temperature was 25°C. A DAD was used and the detection wavelength was at 218 nm. Under the optimum conditions, the baseline separation of seven components was achieved in less than 26 min. Excellent precision, good stability, and accuracy were obtained. For all analytes, linear calibrations were established within 10–100 μg/mL. The LOD and LOQ were within 1.2–4.2 μg/mL and 4.0–14 μg/mL, respectively. The developed method was suitable for the determination of key components in TWHF and TPs.     

Simple method for determination of five terpenoids from different parts of Tripterygium wilfordii and its preparations by HPLC coupled with evaporative light scattering detection

By optimizing the extraction, separation, and analytical conditions, a reliable and accurate high-performance liquid chromatography method coupled with evaporative light scattering detection (ELSD) was developed for simultaneous determination of five terpenoids, i.e., triptolide, tripchlorolide, demethylzelastral, wilforlide B, and wilforlide A, in root, stem, leaves, root bark, twig, and root without bark of Tripterygium wilfordii Hook. f and six of its herbal preparations. This approach would thus provide a more accurate and general method for evaluating the quality of the herb and its preparations. Separation of these five terpenoids was achieved on a ZORBAX Eclipse XDB-C8 column with gradient elution using water and acetonitrile as solvents, both containing 0.05% formic acid, at a temperature of 30 degrees C and a flow rate of 0.8 mL/min. The drift tube temperature of ELSD was set at 100 degrees C, and the nitrogen flow rate at 1.5 L/min. Good linear relationships were obtained with correlation coefficients for the analytes exceeding 0.992, and the LOD and LOQ were less than 0.149 microg and 0.297 microg on column, respectively. Intra-day and inter-day precision of the analytes were less than 1.25% and 5.97%, respectively, and the average recovery rates obtained were in the range of 95.9 +/- 3.7% to 100.4 +/- 5.0% for all terpenoids with RSDs below 4.99%. Quantitative analysis of the five terpenoids in different parts of Tripterygium wilfordii and its six preparations showed that the contents of the terpenoids varied significantly. The tender root contained higher concentrations of triptolide, tripchlorolide, demethylzelastral, and wilforlide B than any other part of the herb. Correspondingly, the root bark contained the greatest concentration of wilforlide A, and the stem and twig came in second and third. This suggested that we could infer whether the medicinal materials were absolute roots without bark or not from the comparative contents of these terpenoids in the tablets in view of the fact that only the roots without bark are the valid officinal part of the plant. This method and the quantitation results obtained can provide a scientific and general as well as simple and convenient approach for the product manufacturers to set up quality control standards and for informing the public about the quality and safety of the preparations.     

Intermittent counter-current extraction as an alternative approach to purification of Chinese herbal medicine

This paper describes intermittent counter-current extraction, a novel method of using a conventional twin column counter-current chromatograph to either split a sample into two groups of compounds or extract and enrich a target compound from a crude extract. The first method is demonstrated by splitting a model mixture of four compounds into two groups. The second method is demonstrated by the extraction and enrichment of a high value target compound, triptolide, from a Chinese herbal medicine crude extract of Tripterygium wilfordii Hook. f., where it is found at low concentration (2%). This was achieved by retaining and enriching the target compound within the column while washing away all other components of the crude material. The success of the first method allowed the second method to be carried out without the need for costly preliminary experiments with the high value sample. 188 mg of triptolide at greater than 98% purity was separated from 9.2 g of crude extract, using 10 l of solvent in a 3-h separation.