Qualitative and quantitative determination of Atractylodes rhizome using ultra‐performance liquid chromatography coupled with linear ion trap–Orbitrap mass spectrometry with data‐dependent processing

A quick and effective workflow based on ultra‐performance liquid chromatography coupled with electron spray ionization and LTQ‐Orbitrap mass spectrometry (UPLC‐LTQ‐Orbitrap MS) was established for compositional analysis and screening of the characteristic compounds of three species of Atractylodes rhizome for quality evaluation. This technique was employed to determine the seven main components in Atractylodes rhizome samples. Ultimately, 78 constituents were identified; of these, seven characteristic compounds were selected for species discrimination, comprising atractylodin (63), atractylenolide I (43), atractylenolide II (49), atractylenolide III (53), atractylon (69), methyl‐atractylenolide II (54) and (4E,6E,12E)‐tetradecadecatriene‐8,10‐diyne‐1,3‐diacetate (59). The seven main compounds, including six characteristic compounds, were simultaneously determined in 29 batches of Atractylodes rhizome samples. Thus, the method validation showed acceptable results. Quantitative analysis showed significantly different contents of the seven main components among the three species of Atractylodes rhizome, which indicates possible distinctions in the pharmacological effects. This established method can simultaneously provide qualitative and quantitative results for compositional characterization of Atractylodes rhizomes and for quality control.     

Quantitative and fingerprinting analysis of Atractylodes rhizome based on gas chromatography with flame ionization detection combined with chemometrics

This study assessed the feasibility of gas chromatography with flame ionization detection fingerprinting combined with chemometrics for quality analysis of Atractylodes rhizome. We extracted essential oils from 20 Atractylodes lancea and Atractylodes koreana samples by hydrodistillation. The variation in extraction yields (1.33–4.06%) suggested that contents of the essential oils differed between species. The volatile components (atractylon, atractydin, and atractylenolide I, II, and III) were quantified by gas chromatography with flame ionization detection and confirmed by gas chromatography with mass spectrometry, and the results demonstrated that the number and content of volatile components differed between A. lancea and A. koreana. We then calculated the relative peak areas of common components and similarities of samples by comparing the chromatograms of A. lancea and A. koreana extracts. Also, we employed several chemometric techniques, including similarity analysis, hierarchical clustering analysis, principal component analysis, and partial least‐squares discriminate analysis, to analyze the samples. Results were consistent across analytical methods and showed that samples could be separated according to species. Five volatile components in the essential oils were quantified to further validate the results of the multivariate statistical analysis. The method is simple, stable, accurate, and reproducible. Our results provide a foundation for quality control analysis of A. lancea and A. koreana.     

Differentiation of essential oils in Atractylodes lancea and Atractylodes koreana by gas chromatography with mass spectrometry

Atractylodes rhizome is a valuable traditional Chinese medicinal herb that comprises complex several species whose essential oils are the primary pharmacologically active component. Essential oils of Atractylodes lancea and Atractylodes koreana were extracted by hydrodistillation, and the yield was determined. The average yield of essential oil obtained from A. lancea (2.91%) was higher than that from A. koreana (2.42%). The volatile components of the essential oils were then identified by a gas chromatography with mass spectrometry method that demonstrated good precision. The method showed clear differences in the numbers and contents of volatile components between the two species. 41 and 45 volatile components were identified in A. lancea and A. koreana, respectively. Atractylon (48.68%) was the primary volatile component in A. lancea, while eudesma‐4(14)‐en‐11‐ol (11.81%) was major in A. koreana. However, the most significant difference between A. lancea and A. koreana was the major component of atractylon and atractydin. Principal component analysis was utilized to reveal the correlation between volatile components and species, and the analysis was used to successfully discriminate between A. lancea and A. koreana samples. These results suggest that different species of Atractylodes rhizome may yield essential oils that differ significantly in content and composition.     

白术煎剂表面增强拉曼光谱分析

测试分析了白术煎剂及其在银胶中的拉曼光谱,并对其进行初步谱峰归属。白术煎剂表面增强拉曼光谱在396,548,617,730,955和1 327 cm-1处,出现6个明显的拉曼信号。测试了白术煎剂和白术煎剂-银胶混合体系的紫外-可见吸收光谱,该吸收光谱在长波区出现新共振吸收峰(999 nm处),进而研究了白术煎剂在银溶胶上的吸附特性及其表面增强机理。结果表明,表面增强拉曼光谱可能为白术煎剂或其他中药煎剂提供一种准确、直接、快速的检测方法。     

Development and Validation of Improved Reversed Phase-HPLC Method for Simultaneous Determination of Curcumin, Demethoxycurcumin and Bis-Demethoxycurcumin

Isocratic reversed phase high performance liquid chromatographic (HPLC) method using RP C18 column was developed for simultaneous determination of the curcuminoids. Mobile phase consisted of acetonitrile:0.1% trifluro-acetic acid (50:50) and flow rate was 1.5 mL min⁻¹ and elution was monitored at 420 nm. Validation in selected conditions showed that the chosen method is sensitive, selective, precise and reproducible with linear response of detector for the simultaneous determination of curcumin (C), demethoxycurcumin (DMC) and bis-demethoxycurcumin (BDMC). The limits of detection were 27.99, 31.91 and 21.81 ng mL⁻¹ for C, DMC and BDMC, respectively. Limits of quantitation for C, DMC and BDMC, were 84.84, 96.72 and 66.10 ng mL⁻¹, respectively. Linear range was form 100 to 600 ng mL⁻¹. The mean ± SD percent recoveries of curcuminoids were 99.87 ± 0.34, 100.09 ± 0.48 and 100.10 ± 0.60% of C, DMC and BDMC, respectively. Further, the method was used for quantitation of curcuminoids from turmeric rhizome.     

气相-质谱分析北细辛根和根茎中的挥发性成分

 采用 质谱联用技术对北细辛根和根茎的挥发性成分进行分离分析 ,鉴定出了 36种化学成分 ,主要成分为十五烷、甲基丁香酚、黄樟醚、3 ,5 二甲氧基甲苯等。对照北细辛根和根茎的分析结果可以发现 ,两者的挥发性成分基本相同 ,但含量有较大差异 ,这为北细辛的临床合理用药及质量控制标准的确定提供了实验依据。     

UPLC-Q-Exactive/MS–based metabonomics revealed protective effect of Zingiberis rhizome and its processed product on deficiency-cold and hemorrhagic syndrome rats

Zingiberis rhizome carbonisata (ZRC) is the processed product of Zingiberis rhizome (ZR). ZR is mainly used for warming the spleen and stomach to dispel cold, whereas ZRC is commonly applied as a treatment for deficiency-cold and hemorrhagic syndrome (DCHS). Although they have long been used to serve different clinical purposes, the specific action mechanism of the drugs and molecular changes underlying ZR processing are not clear. In this study, metabolomics study was carried out to analyze the alterations in endogenous metabolites in serum and urine samples of DCHS rat models using ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry technique and constructed principal component analysis score plots that showed that the ZRC group was completely separated from the DCHS and ZR groups but demonstrated a highly close plotting to the normal control group. The results revealed that both ZR and ZRC intervened in the metabolic pathways of DCHS models but to varying degrees and with different influencing factors. In addition, ZRC was found to function as a treatment for the metabolic disorders of DCHS through 15 pharmacodynamic biomarkers involving a series of pathways, such as glycine, serine, and threonine metabolic pathway, as well as arachidonic acid metabolic pathways. This study showed that metabolomics method based on ultra-high-performance liquid chromatography coupled with quadrupole-Exactive mass spectrometry could preliminarily illuminate the therapeutic mechanism of ZR and ZRC on DCHS and the changes in ZR processing from the molecular-level perspective. The results also provided new insight into further research on DCHS treatment.     

Herbal Composition LI73014F2 Alleviates Articular Cartilage Damage and Inflammatory Response in Monosodium Iodoacetate-Induced Osteoarthritis in Rats

The aim of this study was to determine the anti-osteoarthritic effects of LI73014F2, which consists of Terminalia chebula fruit, Curcuma longa rhizome, and Boswellia serrata gum resin in a 2:1:2 ratio, in the monosodium iodoacetate (MIA)-induced osteoarthritis (OA) rat model. LI73014F2 was orally administered once per day for three weeks. Weight-bearing distribution and arthritis index (AI) were measured once per week to confirm the OA symptoms. Synovial membrane, proteoglycan layer, and cartilage damage were investigated by histological examination, while synovial fluid interleukin-1β level was analyzed using a commercial kit. Levels of pro-inflammatory mediators/cytokines and matrix metalloproteinases (MMPs) in the cartilage tissues were investigated to confirm the anti-osteoarthritic effects of LI73014F2. LI73014F2 significantly inhibited the MIA-induced increase in OA symptoms, synovial fluid cytokine, cartilage damage, and expression levels of pro-inflammatory mediators/cytokines and MMPs in the articular cartilage. These results suggest that LI73014F2 exerts anti-osteoarthritic effects by regulating inflammatory cytokines and MMPs in MIA-induced OA rats.     

FT-Raman光谱法无损鉴定药材的真伪

本文利用傅里叶变换拉曼（FT-Raman）光谱法无损快速鉴定了正品大黄与伪品大黄以及有掺伪物的知母样品。结果表明：大黄（西宁大黄）与伪品大黄（华北大黄，山大黄，水根大黄）在拉曼谱图中均有各自的特征峰，可容易地将它们区别开；知母样品中所掺杂的其他饮片，经与白芨的拉曼谱图比对，证实为白芨饮片，该方法具有快速，准确，操作简单，不需对样品进行提取分离，可直接进行测定的特点。     

Composition and in vitro cytotoxic activities of essential oil of Hedychium spicatum from different geographical regions of western Himalaya by principal components analysis

The rhizome of Hedychium spicatum has been widely used in traditional medicines. The present study deals with the evaluation of the cytotoxic potential of rhizome essential oils from four different regions of the Western Himalaya (India) along with comparative correlation analysis to characterise the bioactive cytotoxic component. The essential oils were coded as MHS-1, MHS-2, MHS-3 and MHS-4, and characterised using GC-FID and GC–MS. The main volatile compounds identified were 1,8-cineol, eudesmol, cubenol, spathulenol and α-cadinol. In vitro cytotoxic activities were assessed against human cancer cell lines such as, the lung (A549), colon (DLD-1, SW 620), breast (MCF-7, MDA-MB-231), head and neck (FaDu), and cervix (HeLa). MHS-4 is significantly active in comparison to other samples against all cancer cell lines. Sample MHS-4 has major proportion of monoterpene alcohol mainly 1,8-cineol. Principal components analysis was performed for the experimental results and all four samples were clustered according to their percentage inhibition at different doses.