Quantification and discovery of quality markers from Toddalia asiatica by UHPLC-MS/MS coupled with chemometrics.

INTRODUCTION: Toddalia asiatica (TA) is a classical traditional Chinese medicine used to treat rheumatoid arthritis and contusions. However, research regarding TA quality control is currently limited. OBJECTIVE: We aimed to establish a strategy for identifying quality markers that can be used for the evaluation of the quality of TA. METHOD: A rapid and efficient ultra-high-performance liquid chromatography coupled with triple quadrupole tandem mass spectrometry (UHPLC-MS/MS) method was developed for the quantitative determination of 19 compounds in TA from different regions. Then, the extraction process of TA was successively optimized by single-factor optimization and response surface methodology. Moreover, chemometrics was employed to confirm the correlation between quality and target compounds. RESULTS: Utilizing the UHPLC-MS/MS method, separation of the 19 bioactive compounds was achieved within 14 min. The method was validated in terms of linearity (r2 > 0.9982), precision (0.08%-3.70%), repeatability (0.50%-2.54%), stability (2.26%-5.46%), and recovery (95.8%-113%). The optimal extraction process (extraction solvent, 65% ethanol aqueous solution; solid-liquid ratio, 1:20; extraction time, 25 min) was determined with the total content of 19 bioactive compounds as indicator. Significant disparities were observed in the contents of target compounds across different batches of TA. Besides, all samples could be categorized into two distinct groups, and magnoflorine, (-)-lyoniresinol, nitidine chloride, norbraylin, skimmianine, and decarine were identified as quality markers. CONCLUSION: In the present study, we developed a strategy to improve the quality control of TA. In consideration of the pharmacodynamic activity and statistical differences, six compounds are proposed as quality markers for TA.

Comparison of chemometric assisted targeted and untargeted approaches for the prediction of radical scavenging activity of ylang-ylang essential oils.

Essential oils have been used for centuries for their preservative properties. An example is ylang-ylang Cananga odorata [Lam.] Hook. f. & Thomson essential oil, which exists in four different distillation grades, where the fraction with the longest distillation time has the highest radical scavenging activity (RSA). Gas chromatography mass spectrometry (GC-MS) followed by multivariate statistical analysis is a powerful approach for determination of RSA. Herein the performance of such multivariate statistical analysis using three data sets derived from gas chromatography mass spectrometry (GC-MS) analysis, is compared to that achieved using two direct and fast spectroscopic techniques, for the prediction of RSA using partial least squares (PLS) regression analysis. The three GC-MS data sets were, 'full chemical composition', 'total chromatogram average mass spectra (TCAMS)' and 'segment average mass spectra (SAMS)', whilst two spectroscopic techniques, namely attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy and Raman spectroscopy, provided the spectroscopic data sets for comparison. PLS models created using ATR-FTIR and 'full chemical composition' data sets provided the lowest relative error of prediction (REP) and mean error of prediction (MEP) in validation, whilst in independent test sets, the PLS models created using ATR-FTIR and SAMS data sets delivered the lowest REP and MEP. The three GC-MS derived data sets were further compared for value in determination of compounds contributing to the RSA. PLS regression analysis of the full chemical composition data set revealed that germacrene D and (E,E)-α-farnesene were the major contributors to the RSA, whilst average mass spectrum based data sets, TCAMS and SAMS, also highlighted eugenol as another contributor to the RSA.

Characteristic profiling of Aconiti Lateralis Radix for distinguishing it from compatible herbal pair using UPLC-Q-TOF-MS coupled with chemometrics.

A method combining ultra-high-performance liquid chromatograph/quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS) and chemometrics was established to evaluate the differences in chemical composition between Aconiti Lateralis Radix (Fuzi in Chinese) before and after combination with Glycyrrhizae Radix et Rhizoma (Gancao in Chinese). UPLC-Q-TOF-MS was used to characterize the chemical components before and after the combination of Fuzi with Gancao, and genetic algorithm selection variables were applied to extract important variables. Partial least square discriminant analysis was used to verify the reliability of the variables obtained by genetic algorithm selection in differentiating Fuzi and combinations with Gancao, and nine potential chemical markers were obtained. The changes in content of chemical markers in Fuzi before and after combination were visualized using a heat map and hierarchical cluster analysis. Based on the chemical markers, characteristic profiling of UPLC-Q-TOF-MS data was developed, then unsupervised principal components analysis and a supervised counter-propagation artificial neural network were used to validate the characteristic profiling approach and showed that it performed well in differentiating between Fuzi and combinations with Gancao.

Investigation on the Q-markers of Bushen Huoxue Prescriptions for DR treatment based on chemometric methods and spectrum-effect relationship.

ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic retinopathy (DR) is a kind of complex complication of late diabetes mellitus with high incidence and risk of blindness. Bushen Huoxue Prescription (BHP), which consists of Rehmanniae radix (RR), Salviae miltiorrhizae radix et rhizoma (SMRR), Ginseng radix et rhizome (GRR) and Puerariae lobatae radix (PLR), has an active effect on the treatment of DR. However, the quality markers (Q-markers) of BHP are not entirely clear. PURPOSE: This study aimed to screen the Q-markers of BHP for DR treatment based on the establishment of spectrum-effect relationship and verified experiment. MATERIALS AND METHODS: In this study, 12 BHP samples (S1-S12) for fingerprint analysis and pharmacological evaluation were prepared according to a four-factor and twelve-level uniform design. High performance liquid chromatography-ultraviolet detector-evaporative light scattering detector (HPLC-UV-ELSD) was employed to analyze the fingerprint on the basis of the characteristics of BHP components. The evaluation of sample similarity was carried out by similarity analysis (SA) and hierarchical cluster analysis (HCA). The pharmacological indicators, including expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) in the retina of Sprague Dawley (SD) rats induced by streptozotocin (STZ), were detected by enzyme-linked immunosorbent assay (ELISA). Besides, the spectrum-effect relationship between common peaks of fingerprints and the pharmacological results was investigated by partial least squares regression (PLSR) and canonical correlation analysis (CCA). The results of spectrum-effect relationship were verified by the expression of VEGF and HIF-1α on primary culture retinal Müller cells induced by hyperglycemia and hypoxia. RESULTS: In the HPLC-UV-ELSD fingerprint, 23 common peaks in UV and 14 common peaks in ELSD were identified. The pharmacological results indicated that the expression of VEGF and HIF-1α in the retina of SD rats was inhibited by 12 BHP samples to varying degrees compared with the model group. Based on SA and heatmap of HCA, S4 and S8 were clearly distinguished from other samples. The results of PLSR and CCA revealed that the contents of puerarin, daidzin, salvianolic acid B and ginsenoside Rb1 were inversely correlated with the expression of VEGF and HIF-1α. Hence, the four compounds may be the main active components to prevent and treat DR. The results of intervention on primary culture retinal Müller cells showed that puerarin, daidzin, salvianolic acid B, and ginsenoside Rb1 can significantly inhibit the expression of VEGF and HIF-1α. CONCLUSIONS: The spectrum-effect relationship of BHP was successfully established, and the Q-markers of BHP for the prevention and treatment of DR were preliminarily confirmed. It provides a feasible method for the research of quality control.

Cancer Influences the Elemental Composition of the Myocardium More Strongly than Conjugated Linoleic Acids-Chemometric Approach to Cardio-Oncological Studies.

The aim of the study was to verify in a cardio-oncological model experiment if conjugated linoleic acids (CLA) fed to rats with mammary tumors affect the content of selected macro- and microelements in their myocardium. The diet of Sprague-Dawley females was supplemented either with CLA isomers or with safflower oil. In hearts of rats suffering from breast cancer, selected elements were analyzed with a quadrupole mass spectrometer with inductively coupled plasma ionization (ICP-MS). In order to better understand the data trends, cluster analysis, principal component analysis and linear discriminant analysis were applied. Mammary tumors influenced macro- and microelements content in the myocardium to a greater extent than applied diet supplementation. Significant influences of diet (p = 0.0192), mammary tumors (p = 0.0200) and interactions of both factors (p = 0.0151) were documented in terms of Fe content. CLA significantly decreased the contents of Cu and Mn (p = 0.0158 and p = 0.0265, respectively). The level of Ni was significantly higher (p = 0.0073), which was more pronounced in groups supplemented with CLA. The obtained results confirmed antioxidant properties of CLA and the relationship with Se deposition. Chemometric techniques distinctly showed that the coexisting pathological process induced differences to the greater extent than diet supplementation in the elemental content in the myocardium, which may impinge on cardiac tissue's susceptibility to injuries.

A metabolomics approach to evaluate the effect of lyophilization versus oven drying on the chemical composition of plant extracts.

Lyophilization is the "gold standard" for drying plant extracts, which is important in preserving their quality and extending their shelf-life. Compared to other methods of drying plant extracts, lyophilization is costlier due to equipment, material and operational expenses. An alternative method is post-extraction oven-drying, but the effects of this process on extract quality are unknown. In this study, crude extracts from Arthrocnemum macrostachyum shoots were compared using three post-extraction drying methods (lyophilization and oven drying at 40 and 60 °C) and two extraction solvents (water and aqueous 50% ethanol). Untargeted metabolomics coupled with chemometrics analysis revealed that post extraction oven-drying resulted in the loss of up to 27% of molecular features when compared to lyophilization in water extracts only. In contrast, only 3% of molecular features were lost in aqueous 50% ethanol extracts when subjected to oven drying. That is to say, ethanol used as a solvent has a stabilizing effect on metabolites and enhances their resistance to thermal transformation in the oven. Collectively, oven-drying of extracts was as effective as lyophilization in preserving metabolites in extracts only when 50% ethanol was used as a solvent. The results presented in this paper demonstrate the value of selecting solvent-appropriate post-extraction drying methods.

Chemometric approach to find relationships between physiological elements and elements causing toxic effects in herb roots by ICP-MS.

In this paper 13 elements, both physiological and causing toxic effects, were determined by inductively coupled plasma mass spectrometry in roots of 26 species of herbs used in Traditional Chinese Medicine. The herbs were purchased from online shop in two batches 1 year apart to verify the variability of elemental content in time. The multivariate statistical methods-multiple regression, canonical variates and interaction effect analysis-were applied to interpret the data and to show the relationships between elements and two batches of herb roots. The maximum permissible concentration of Cd (0.3 mg kg-1) was exceeded in 7 herb roots which makes 13% of all specimens. The multiple regression analysis revealed the significant relationships between elements: Mg with Sr; V with Pb, As and Ba; Mn with Pb; Fe with As and Ba; Co with Ni and Sr, Cu with Pb, Cd and As; Zn with Pb, Cd, As and Ba. The canonical variates analysis showed that the statistical inference should not be based solely on the type of herb or number of batch because of the underlying interaction effects between those two variables that may be a source of variability of the content of determined elements.

Quality assessment of Succus Bambusae oral liquids based on gas chromatography/mass spectrometry fingerprints and chemometrics.

RATIONALE: Succus Bambusae is consumed as a kind of herbal medicine and natural beverage in China. However, the current quality standards for Succus Bambusae are low and lack safety indicators, which makes it difficult to effectively guarantee its quality. Therefore, it is of great significance to study the identification and quality control technology for the product. METHODS: We have developed a set of qualitative and quantitative methods based on gas chromatography/mass spectrometry (GC/MS) for the analysis of volatile components in Succus Bambusae oral liquid (SBOL). Combining GC/MS fingerprint analysis and related chemometrics algorithms, with similarity evaluation, Hotelling T2 and distance to Model X (DModX) as criteria, the quality consistency of different batches was evaluated, and SBOL samples from different manufacturers were differentiated. RESULTS: Twenty-nine volatile components were preliminarily identified from 40 batches of SBOL samples from six manufacturers, and six Q-markers (Quality Markers) for the SBOLs were discussed and determined using GC/MS. The products from different manufacturers were distinguished using chemometrics. CONCLUSIONS: The results showed that the quality of the SBOL samples from different batches and different manufacturers fluctuated greatly, which suggested that research into the raw materials and manufacturing techniques should be strengthened to improve the quality of SBOL and ensure its quality consistency.

Mineral Composition of Dietary Supplements-Analytical and Chemometric Approach.

There is a lack of data on the actual composition and effectiveness of beetroot-based dietary supplements. The research aimed to determine the profile of 22 elements (Na, K, Ca, Mg, P, Fe, As, Se, Zn, Cu, Ag, Co, Ni, Mo, Al, Mn, Sr, Cr, Ba, Li, Pb, Cd) in beetroot and its supplements by the microwave plasma atomic emission spectrometry (MP-AES) method. The analytical procedure was optimised and validated. The composition of both groups was compared, assessing compliance with the recommended daily doses for the chosen elements, and the health risk was estimated. Furthermore, chemometric analysis was applied. Beetroots constituted a significant source of elements, especially K, Na, Mg, Ca, P, in contrast to supplements which contained their negligible amounts except from iron-enriched products which provided notable amounts of Fe (38.3-88% of the Recommended Dietary Allowance for an adult male from 19 to 75 years old). Some products were significantly contaminated with toxic elements (As, Cd). Factor and cluster analyses were helpful in the differentiation of beetroot and its supplements in view of their type (vegetable, supplement, iron-enriched supplement), origin, type of cultivation (conventional, organic), and form (capsule, tablet) based on their mineral composition. The obtained results indicate the need for more stringent control of supplements, as they may pose a significant health risk to consumers.