High-performance Liquid Chromatography Coupled With Evaporative Light Scattering Detection Research Articles

Changes in Triacylglycerols Content and Quality Control Implications of Coix Seeds during Processing and Storage.

Coix seed is a kind of widespread cereal, and it is used as a folk medicine in China. The present work focuses on the analysis of changes in triacylglycerols (TAGs) content and mycotoxins of coix seed during the processing and storage period for its quality control. Using ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF-MS) and high-performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD) techniques, 42 lipid components in coix seeds were identified, and seven molecular species of TAG in coix seeds from different localities in China were measured and compared, respectively. A correlation analysis between the morphological features and TAGs contents revealed the integrity instead of the particle size of the seed, displaying a highly positive correlation with its quality. The higher contents of TAGs in hulled coix seed than in polished coix seed proposed an alternative processing way. During storage, the changes in TAGs contents of seeds indicated that the storage period should be less than 3 months, and the intact seeds could maintain lipid stability better than the powder. Furthermore, the air humidity and temperature should be controlled during coix seed storage to prevent the production of mycotoxins. These results provide significant insight into the effective control of coix seed quality during processing and storage.

A strategy for herbal interaction between Ziziphi spinosae Semen and vinegar processed Schisandrae Chinensis fruit when co-decocted based on process route design of new drug of Chinese herbal formulae

For characterizing the influence of vinegar processed Schisandrae Chinensis Fruit (vSCF) on the dissolution of constituents of Ziziphi spinosae Semen (ZSS) when co-decocted, a strategy for herbal interaction was currently proposed based on process route design of new drug of Chinese herbal formulae. Firstly, the content of jujuboside A (JuA) in ZSS-vSCF co-decoction was determined at different ratios by high performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD), to demonstrate the influence of vSCF on JuA of ZSS. As the results showed, the content of JuA was declined drastically in ZSS-vSCF co-decoction compared to the ZSS single decoction, especially at the ratio of 1:1. Hence, the ratio of 1:1 was used in the following studies. Furtherly, as pre-extraction is usually taken in the pharmaceutical industry of Chinese patent medicines (CPM) to avoid herbal interaction, thus the pre-extracted solvents of vSCF were investigated. The content of JuA in coupled decoction was indeed improved when vSCF was pre-extracted before co-decocted with ZSS. Among the pre-extracted solvents, 60% ethanol possessed a better-improved effect. Therefore, 60% ethanol extract of vSCF was treated with JuA and the transformed compounds of JuA can be identified by ultra high performance liquid chromatography with electrospray ionization quadrupole time-of-flight mass spectrometry (UPLC-ESI-Q-TOF-MS/MS). Then, we reported that jujubosides, the main saponins in ZSS, would be transformed into its ebelin-lactone forms through dehydration, RDA cleavage and hydrolysis of glycosidic bond, which could be induced by organic acid in vSCF. Additionally, to elucidate the comprehensive influence, 45 compounds were tentatively identified by UPLC–ESI-Q-TOF-MS/MS, and their dissolution variation in different ZSS-vSCF co-decoction was represented by manually acquired EICs, as well. It can be found that vSCF can promote extraction efficiencies of alkaloids and flavonoid O-glycosides of ZSS when co-decocted, while saponins were suppressed. Similarly, the pre-extraction of vSCF could reverse the suppression. To further elucidate the significance of ZSS-vSCF co-decoction, a comparison of molecular modeling between JuA and its ebelin-lactone form were carried out, revealing that the corresponding ebelin-lactone form had a stronger binding affinity than JuA toward calmodulin (CaM). Collectively, to lower the herbal interaction, a pre-extraction method was developed against the decrease of JuA in ZSS-vSCF co-decoction. And the dissolution of different types of constituents in ZSS was investigated by a semi-quantitative method, as well. Also, ebelin-lactone form of JuA, which resulted from ZSS-vSCF co-decoction, demonstrated a stronger binding affinity toward CaM in virtual screening, but the future pharmacological and pharmacokinetic research is worthy to be further studied.

Application of ATR-FTIR spectroscopy for profiling of non-structural carbohydrates in onion (Allium cepa L.) bulbs

Qualitative and quantitative composition of non-structural carbohydrates comprising glucose, fructose, sucrose and fructooligosaccharides (FOS) is one of the key determinants of market suitability, storability and technological processability of onions. To develop a cost-effective and rapid tool for carbohydrate profiling, applicability of attenuated total reflectance Fourier-transform infrared (ATR-FTIR) spectroscopy of onion juice was investigated with special regard to FOS patterns. As reference, detailed carbohydrate profiles of onion juices were generated by high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD).Hierarchical cluster analysis (HCA) of ATR-FTIR spectra was successfully applied for classifying onions into fresh market, storage and dehydrator type according to HPLC-ELSD profiles.A bootstrapping method for automatized test-set validation by projection to latent structures (PLS) algorithms using HPLC and ATR-FTIR spectroscopy data was developed. Model statistics showed promising perspectives for reliable quantification of individual saccharides and sum parameters. The presented methodology allows estimating the nutritional and pre-biotic value directly during cultivation and processing.

Ginseng ameliorates exercise-induced fatigue potentially by regulating the gut microbiota.

The therapeutic effects of water extract of ginseng (WEG) on exercise-induced fatigue (EF) have been reported in several previous studies, but the molecular mechanisms involved remain unexplored. In this study, the anti-EF effects of WEG were studied, and the potential mechanisms were discussed. We characterized the chemical components of WEG by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) and high performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD), and then examined the anti-EF effects of WEG on a rat model of weight-loaded swimming with a focus on endogenous metabolism and gut microbiota. WEG contains abundant (90.15%, w/w) saccharides and ginsenosides with structurally diverse glycosyls. WEG taken orally showed strong anti-EF effects by ameliorating energy metabolism abnormality, oxidative stress, lipid peroxidation, inflammatory response, disorders in the metabolism of bile acid, amino acid, fatty acid and lipid, as well as the gut microbiota dysbiosis. Given that gut microbiota is significantly associated with energy expenditure, systemic inflammation and host metabolism, these findings suggest a potential central role of the gut microbiota in mediating the anti-EF effect of WEG. That is, the saccharides and ginsenosides in WEG serve as energy substrates for specific intestinal bacteria, thereby beneficially regulating the gut microbiota, and the reshaped gut microbial ecosystem then triggers several molecular and cellular signaling pathways (e.g. butyrate or TGR5 signals) to achieve the therapeutic effects on EF. The outcomes highlighted here enable deeper insight into how WEG overcomes EF.

Characterization of a recombinant bile salt hydrolase (BSH) from Bifidobacterium bifidum for its glycine-conjugated bile salts specificity

Deconjugation of bile salts in vivo not only promotes the cholesterol metabolism of the host, but also provides cellular carbon, nitrogen and sulphur for some probiotics. In this study, the gene of BSH from Bifidobacterium bifidum (ATCC 29521) was amplified and expressed using glutathione S-transferase (GST) Gene Fusion System. The enzyme was purified in a mild conditions without denaturation and renaturation, and the molecular mass was estimated to be 35 kDa. The optimum pH for the enzyme catalysis was 5.5. The BSH enzyme was stable from 37 to 57 °C. In addition, this enzyme was completely inhibited by Cu2+, while many other metal ions only gave moderate inhibition. Hydrolysis efficiency of BSH was quantified in two ways: ninhydrin colouration method and high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD). The hydrolysis process was clearly observed through real-time picture monitoring. The activity of BSH on glycine-conjugated bile salts was better than on taurine-conjugated bile salts. The proportion of hydrolysis for both sodium glycochenodeoxycholate (GCDCA) and sodium glycoursodeoxycholate (GUDCA) were more than 90% in 60 min. However, the proportion of hydrolysis for sodium taurochenodeoxycholate (TCDCA) and sodium tauroursodeoxycholate (TUDCA) were only 68% and 43% at the same condition. The study of this paper provides a better choice for hydrolysis of glycine-conjugated bile salts. Highlights The specificity of BSH was high for glycine-conjugated bile salts. Hydrolysis process of bile acid was obtained through real-time picture monitoring. Hydrolysis efficiency of BSH was quantified in two ways: ninhydrin colouration method and HPLC-ELSD.

Comparative Analysis of Carbohydrates, Nucleosides and Amino Acids in Different Parts of Trichosanthes kirilowii Maxim. by (Ultra) High-Performance Liquid Chromatography Coupled with Tandem Mass Spectrometry and Evaporative Light Scattering Detector Methods.

Trichosanthes kirilowii Maxim. is one of the original plants for traditional Chinese medicines Trichosanthis Fructus, Trichosanthis Semen, Trichosanthis Pericarpium and Trichosanthis Radix. Amino acids, nucleosides and carbohydrates are usually considered to have nutritional value and health-care efficacy. In this study, methods involving high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD), UV-visible spectrophotometry and ultra-high-performance liquid chromatography coupled with tandem mass spectrometry (UHPLC-MS/MS) were established for quantifying carbohydrates (fructose, glucose, stachyose, raffinose and polysaccharide), fourteen nucleosides and twenty one amino acids. Moreover, sixty-three samples from nine different parts, including pericarp, seed, fruit pulp, stem, leaf, main root, main root bark, lateral root and lateral root bark of T. kirilowii from different cultivated varieties were examined. The established methods were validated with good linearity, precision, repeatability, stability, and recovery. The results showed that the average content of total amino acids in roots (15.39 mg/g) and root barks (16.38 mg/g) were relatively higher than for others. Contents of nucleosides in all parts of T. kirilowii were below 1.5 mg/g. For carbohydrates, fruit pulp has a higher content than others for glucose (22.91%), fructose (20.63%) and polysaccharides (27.29%). By using partial least-squared discriminate analysis (PLS-DA), Variable importance in the projection (VIP) plots and analysis of variance (ANOVA) analysis, the characteristic components of the different organs (fruit, stems and leaves, roots) were found. This analysis suggested there were potential medicinal and nutritive health care values in various parts of the T. kirilowii, which provided valuable information for the development and utilization of T. kirilowii.

Stronger anti-obesity effect of white ginseng over red ginseng and the potential mechanisms involving chemically structural/compositional specificity to gut microbiota

BackgroundGinseng has therapeutic potential for treating obesity and the associated gut microbiota dysbiosis. However, whether white ginseng and red ginseng, the two kinds of commonly used processed ginseng, possess different anti-obesity effects remains unknown. PurposeAnti-obesity effects of water extracts of white ginseng and red ginseng (WEWG and WERG) were compared, and the potential mechanisms were discussed. MethodsChemical profiles of WEWG and WERG were characterized by ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) and high performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD). Anti-obesity effects of WEWG/WERG were examined by determining fat accumulation, systemic inflammation, enteric metabolic disorders and gut microbiota dysbiosis in high-fat diet (HFD)-fed obese mice. ResultsBoth WEWG and WERG exerted anti-obesity effects, with WEWG stronger than WERG. Compared to WERG, WEWG contained less contents of carbohydrates (polysaccharides, oligosaccharides, free monosaccharides) and ginsenosides, but chemical structures or compositions of these components in WEWG were characteristic, i.e. narrower molecular weight distribution and higher molar ratios of glucose residues of polysaccharides; higher content ratios of oligosaccharides DP2–3 (di-/tri-saccharides)-to-oligosaccharides DP4–7 (tetra-/penta-/hexa-/hepta-saccharides), sucrose-to-melibiose, maltose-to-trehalose and high-polar-to-low-polar ginsenosides. WEWG better ameliorated fat accumulation, enteric metabolic disorders and gut microbiota dysbiosis in HFD-fed obese mice than WERG. ConclusionThe stronger anti-obesity effect of white ginseng appears to correlate with differences in its chemical profile as compared to red ginseng. The carbohydrates and ginsenosides in WEWG potentially present more structural and compositional specificity to the obesity-associated gut bacteria, allowing more beneficial effects of WEWG on the gut microbiota dysbiosis. This consequently better alleviates the enteric metabolic disorders and systemic inflammation, thereby contributing to the stronger anti-obesity effect of WEWG as compared to WERG.

Comparison of the Bioactive Components in Two Seeds of Ziziphus Species by Different Analytical Approaches Combined with Chemometrics.

The Ziziphus species are considered to be the medicine and food dual purposes plants. Among them, the seed of Ziziphus jujuba var. spinosa (ZS) has traditionally been used as an ethnomedicine in Asian countries for thousands years. Owing to the significant benefits for human health, the demand for ZS increased year by year, and the wild resources have become increasingly scarce, which resulted in a shortage of market supply for ZS and product adulteration by substituting ZS with the seeds of Z. mauritiana Lam. (ZM). However, whether the bioactivity of ZM is similar to ZS has not been fully confirmed till now. Thus, to provide potential information for evaluating the similarity of the health promoting activities between these two Ziziphus seeds, their chemical profiles, including triterpenoids, flavonoids, nucleosides, free amino acids and fatty acids were compared using high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD), ultra-high performance liquid chromatography coupled with triple-quadrupole mass spectrometry (UHPLC-TQ MS), and gas chromatography coupled with mass spectrometry (GC-MS) methods. Furthermore, a more holistic investigation was performed with multivariate principle component analysis and orthogonal projections to latent structures-discriminant analysis analyses to explore the relative variability between the seeds of two species. The results showed that a significant difference exists between ZS and ZM, and ZS was more rich in saponins, polyunsaturated fatty acids and some amino acids, whereas ZM was particularly rich in saturated fatty acids and flavonoids. The above results suggested the bioactivities of ZM for human health may not be similar to ZS owing to their difference in chemical profiles. These results would also be helpful for distinguishing the ZM from ZS with the chemical markers obtained from the study, and set a scientific foundation for establishing the quality control method of ZS.

Determination of Five Aminoalcohol-diterpenoid Alkaloids in the Lateral Root of Aconitum carmichaeli by HPLC-ELSD with SPE.

A convenient and accurate high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) method using solid phase extraction (SPE) was established for quantification of five aminoalcohol-diterpenoid alkaloids (ADAs), including mesaconine, aconine, hypaconine, fuziline and neoline, in the lateral roots of Aconitum carmichaeli (Fuzi) for the first time. The Fuzi extractive was purified using strong cation-exchange SPE. The chromatographic separation was performed on a Gemini C18 column (150 × 4.60 mm, 5 μm) with the mobile phase of methanol-water-diethylamine (48:52:0.01, v/v/v), adjusted to pH 10.2 with acetic acid. The detector was Alltech ELSD 2000ES (drift tube temperature: 90°C; gas flow-rate: 2.3 L/min). Five ADAs in Fuzi were well separated. The calibration curves showed good linearity (r > 0.9990) in the range of 0.0125-0.3750 mg/mL for each alkaloid. The recoveries were in the range of 95.1-105.7%, with relative standard deviations < 5.0%. This method is accurate, specific and repeatable for the determination of five ADAs in different Fuzi samples, which can be applied to the quality control of Fuzi.

Three-Dimensional Evaluation on Ecotypic Diversity of Traditional Chinese Medicine: A Case Study of Artemisia annua L.

Artemisinin is the first-line drug for anti-malaria recommended by the World Health Organization (WHO). As the sole natural plant source of artemisinin, ecotypes of Artemisia annua L. vary widely in artemisinin content between nations, and China is the main producing area of A. annua. Here we present a three-dimensional evaluation on ecotypic diversity of A. annua from 12 main producing areas in China using high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) method, DNA barcoding and ecological analyses. The results indicated that A. annua exhibited high ecotypic diversity. A. annua grown in the South of the Qinling Mountains-Huaihe River Line had a high artemisinin content, whereas the northern ones were low. Similar pattern was noted in the genetic diversity. The southern A. annua had high intraspecific variation in contrast to the northern A. annua. In terms of ecological analyses, humidity and sunshine time could be the major limiting ecological factors that affect the accumulation of artemisinin. This is the first reported three-dimensional evaluation integrating chemical, molecular and ecological analyses of the ecotypic diversity of A. annua. The work will facilitate exploring the genetic basis of chemical variations and developing strategies for the breeding and cultivation of high quality A. annua.

Simultaneous Determination of Four Triterpenoid Saponins in Aralia elata Leaves by HPLC‐ELSD Combined with Hierarchical Clustering Analysis

Aralia elata leaves are known to have several biological activities, including anti-arrythmia, antitumor, anti-inflammatory, anti-fatigue, antimicrobial and antiviral effects. Our previous study found that triterpenoid saponins from the leaves of A. elata had antitumor effects. Quantification of the triterpenoids is important for the quality control of A. elata leaves. To establish high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) for the simultaneous determination of four major triterpenoid saponins, including Aralia-saponin IV, Aralia-saponin VI, 3-O-β-d- glucopyranosyl-(1→3)-β-d-glucopyranosyl-(1→3)-β-d-glucopyranosyl oleanolic acid 28-O-β-d-glucopyranoside (Aralia-saponin TTP)and Aralia-saponin V. The separation was carried out on a Dikma Diamonsil C18 column (4.6mm×250mm, 5μm) efficiently with gradient elution consisting of acetonitrile and water. All calibration curves showed good linear regression (R2 >0.9996) within the ranges of tested concentrations. This validated method was applied to determine the contents of the four major triterpenoid saponins in 53 samples from different regions of northeast China. Hierarchical clustering analysis was first used to classify and differentiate Aralia elata leaves. The method developed was successfully applied to analyse four major triterpenoid saponins in Aralia elata leaves which is helpful for quality control of the herb. Copyright © 2017 John Wiley & Sons, Ltd.

Full component analysis of Tianma-Gouteng-Yin.

BackgroundTianma-Gouteng-Yin (TGY), which is common Chinese medicine formulation consisting of 11 different herbs and being used in China for the treatment of Parkinson’s disease, inflammatory conditions and cardiovascular diseases, was selected for full component analysis. The aim of this study was to quantitatively analyze the chemical profiles of ten commercial TGY samples and one sample produced in our laboratory.MethodsUltra-high performance liquid chromatography (UHPLC) coupled with quadrupole-tandem time-of-flight mass spectrometry (Q-TOF-MS) was used to analyze the non-saccharide small molecule components of the different TGY samples. The established method was validated in terms of its linearity, sensitivity, precision, accuracy and stability. High performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) was also used to quantify three major saccharides (fructose, glucose and sucrose).ResultsThe relative standard deviations for the precision, repeatability and stability of these compounds were less than 5 %, while the accuracy of the method was 95–105 %. Twenty-eight of the compounds found in TGY were successfully identified, with 20 being quantified. The macromolecules present in these samples were also identified using an ethanol precipitation method, representing 294.68–696.64 mg/g of the total material depending on the batch. Notably, the components identified using this method represented up to 78 % of the total weight of the TGY samples.ConclusionsThe developed UHPLC/Q-TOF-MS and HPLC-ELSD methods successfully identified 28 of the complex compounds found in TGY.

Integrating qualitative and quantitative characterization of Prunellae Spica by HPLC-QTOF/MS and HPLC-ELSD

The present study was designed to analyze the major constituents in Prunellae Spica and establish a method for simultaneous determination of two constituents contained in Prunellae Spica. High performance liquid chromatography coupled with time-of-flight mass spectrometry (HPLC-QTOF-MS/MS) technique was used to identify the constituents in the extractive of Prunellae Spica. High performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) was used to simultaneously quantify two kinds of constituents contained in Prunellae Spica. Principal component analysis (PCA) was applied to compare the similarity and difference among samples from different regions of China. In the present study, 22 compounds were identified and some new fragmental pathways of triterpenic acids were discovered. An accurate and reliable HPLC-ELSD method was developed and validated for the first time to simultaneously quantify multiple constituents, including rosmarinic acid, maslinic acid, corosolic acid, betulin, oleanolic acid, and ursolic acid in the extract of Prunellae Spica. (PCA) revealed some similarities and differences among different samples from different regions of China. In conclusion, our results from this study would be helpful in establishing a scientific and rational quality control method for Prunellae Spica.

Simultaneous Determination of Two Major Triterpenoid Saponins: Celosins I and II in Celosiae Semen by HPLC-ELSD

ObjectiveTo establish an analytical method for the simultaneous determination of celosin I and celosin II, two major triterpenoids in Celosia Semen and compare the contents of celosin I and celosin II from different habitats to screen the resources of elite germplasm for further applications. MethodsA sensitive and simple high-performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD) method was developed for the first time for the simultaneous determination of celosin I and celosin II. Using this method, 21 batches of Celosiae Semen were determined from different habitats in China. ResultsThere was an obvious difference in the contents of celosin I and celosin II among Celosiae Semen species from various habitats across China. The crude drug from Yongzhou, Hunan province, Zhuhai, Guangdong province, and Nanning, Guangxi province showed the highest contents of all habitats, while Anguo, Hebei province, Haidian, Beijing, and Zhengzhou, Henan province showed the lowest content. The results also showed that geographical location had a great influence on the contents of the two compounds. The batches from lower latitudes were higher in contents of celosin I and celosin II. ConclusionThe sun light may be a key factor influencing the contents of the two saponins, indicating that the environment has a great impact on the quality of Celosiae Semen.

P-453 – Évaluation des pratiques de prévention de la douleur du prématuré

To establish an analytical method for the simultaneous determination of celosin I and celosin II, two major triterpenoids in Celosia Semen and compare the contents of celosin I and celosin II from different habitats to screen the resources of elite germplasm for further applications.A sensitive and simple high-performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD) method was developed for the first time for the simultaneous determination of celosin I and celosin II. Using this method, 21 batches of Celosiae Semen were determined from different habitats in China.There was an obvious difference in the contents of celosin I and celosin II among Celosiae Semen species from various habitats across China. The crude drug from Yongzhou, Hunan province, Zhuhai, Guangdong province, and Nanning, Guangxi province showed the highest contents of all habitats, while Anguo, Hebei province, Haidian, Beijing, and Zhengzhou, Henan province showed the lowest content. The results also showed that geographical location had a great influence on the contents of the two compounds. The batches from lower latitudes were higher in contents of celosin I and celosin II.The sun light may be a key factor influencing the contents of the two saponins, indicating that the environment has a great impact on the quality of Celosiae Semen.

P-474 – Neurofibromatose type I (NF1): une présentation clinique particulière

To establish an analytical method for the simultaneous determination of celosin I and celosin II, two major triterpenoids in Celosia Semen and compare the contents of celosin I and celosin II from different habitats to screen the resources of elite germplasm for further applications.A sensitive and simple high-performance liquid chromatography coupled with evaporative light-scattering detector (HPLC-ELSD) method was developed for the first time for the simultaneous determination of celosin I and celosin II. Using this method, 21 batches of Celosiae Semen were determined from different habitats in China.There was an obvious difference in the contents of celosin I and celosin II among Celosiae Semen species from various habitats across China. The crude drug from Yongzhou, Hunan province, Zhuhai, Guangdong province, and Nanning, Guangxi province showed the highest contents of all habitats, while Anguo, Hebei province, Haidian, Beijing, and Zhengzhou, Henan province showed the lowest content. The results also showed that geographical location had a great influence on the contents of the two compounds. The batches from lower latitudes were higher in contents of celosin I and celosin II.The sun light may be a key factor influencing the contents of the two saponins, indicating that the environment has a great impact on the quality of Celosiae Semen.

A novel strategy for rapid quantification of 20(S)-protopanaxatriol and 20(S)-protopanaxadiol saponins in Panax notoginsengP. ginseng and P. quinquefolium

A novel strategy for the qualitative and quantitative determination of 20(S)-protopanaxatriol saponins (PTS) and 20(S)-protopanaxadiol saponins (PDS) in Panax notoginseng, Panax ginseng and Panax quinquefolium, based on the overlapping peaks of main components of PTS (calibrated by ginsenoside Rg1) and PDS (calibrated by ginsenoside Rb1), was proposed. The analysis was performed by using high-performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD). Under specific chromatographic conditions, all samples showed two overlapping peaks containing several main ginsenosides belonging to PTS and PDS, respectively. The overlapping peaks were also identified by using HPLC–MS. Based on the sum and ratio of PTS and PDS, 60 tested Panax samples were divided into three main clusters according to their species. The findings suggested that this strategy provides a simple and rapid approach to quantify PTS and PDS in Panax herbs.

Comparing major alkaloids of Fritillariae Hupehensis Bulbs (FHB) and congeneric plants by HPLC-ELSD and HPLC-ESI-MSn

In the study, the major alkaloids from Fritillariae Hupehensis Bulbs (FHB) have been analysed for the first time by the combined use of the following two methods: the simultaneous quantitation of three alkaloids by using high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) and the simultaneous characterisation of seven alkaloids by using HPLC coupled with electrospray ionisation-mass spectrometry analysers detection (HPLC-ESI-MSn). The other four congeneric species were compared by using the established method. Both correlation coefficients of similarity in chromatograms were calculated for quantitative expression of HPLC profiles. The results revealed that the chromatogram profile combining similarity evaluation could efficiently identify and distinguish FHB from other different origins of Fritillaria species. The established method was considered to be suitable for checking the genuine origin and quality control of FHB.

Quality control and identification of steroid saponins from Dioscorea zingiberensis C. H. Wright by fingerprint with HPLC-ELSD and HPLC-ESI-Quadrupole/Time-of-fight tandem mass spectrometry

In this study, a fingerprint of steroid saponins, the major bioactive constituents in the crude extracts from Dioscorea zingiberensis C. H. Wright (DZW), has been established for the first time by combined use of the following two methods: high-performance liquid chromatography coupled with evaporative light scattering detector (HPLC-ELSD) and the simultaneous characterization of the steroid saponins by high-performance liquid chromatography coupled with electrospray ionization-mass spectrometry and quadrupole tandem time-of-fight mass analyzers detection (HPLC-ESI-Q/TOF). All HPLC analyses were carried out on a Welchrom C18 column (250mm×4.6mm I.D., 5μm) with a mobile phase composed of water and acetonitrile under gradient elution. There were 68 common characteristic peaks in the fingerprints, in which 12 of them were confirmed by comparing their mass spectra and retention times with those of the reference compounds. In order to identify other unknown peaks, their fragmentation behaviors characteristic of the major groups of steroid saponins from DZW with six types of aglycone skeletons were discussed in detail, and possible MS/MS fragmentation pathways were proposed for aiding the structural identification of these components. According to the summarized fragmentation patterns, these peaks were tentatively assigned by matching their empirical molecular formula with those of the published compounds, or by elucidating their quasi-molecular ions and fragment ions referring to available literature information when the reference standards were unavailable. As a result, 22 new steroid saponins were found in DZW for the first time. In addition, the quantitative analysis of the nine (except for the reference compounds A, B, and C) known peaks was accomplished at the same time which indicated that there was a great variability in the amount of these active compounds in different batches in the crude extracts. This approach could demonstrate that the fingerprint could be considered to be a suitable tool to comprehensively improve the quality control of DZW. The identification and structural elucidation of the peaks in the fingerprint may provide important experimental data for further pharmacological and clinical researches.

Simultaneous Determination of Flavonoids and Saponins in Semen Ziziphi Spinosae (Suanzaoren) by High Performance Liquid Chromatography with Evaporative Light Scattering Detection

AIM: To simultaneously determinate the six major components in Semen Ziziphi Spinosae (Suanzaoren) by high performance liquid chromatography coupled with evaporative light scattering detection (HPLC-ELSD) for a scientific evaluation of its quality. METHOD: The separation was achieved on a CLC-ODS C18 analytical column. The mobile phase consisted of 0.1% aqueous acetic acid (A) and acetonitrile (B) using a gradient elution. The drift tube temperature of ELSD was set at 106.5℃, and nitrogen flow-rate was 2.9 L•min^(-1). RESULT: Three flavonoids (isovitexin-2”-O-β-glucopyranoside, spinosin and 6'”-feruloylspinosin) and three saponins (jujuboside A, jujuboside B and jujuboside D) showed good linearity (r^2＞0.999) within test ranges. This method was well validated with good precision, repeatability and sensitivity. It was successfully applied to quantify three flavonoids and three saponin components in twenty-five samples. CONCLUSION: The developed simple, rapid and reliable HPLC-ELSD method is suitable for the comprehensive quality control of Suanzaoren as well as its preparations.