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.

Preparation, optimization, characterization and in vitro release of baicalein-solubilizing glycyrrhizic acid nano-micelles.

Glycyrrhizic acid is an amphiphilic molecule, which can form host-guest complexes by self-assembly, thereby encapsulating the guest molecule and increasing its solubility. The complexes can also achieve a controlled release effect for encapsulated drugs, so they have potential as drug delivery-systems. Baicalein is a flavonoid, with many pharmacological activities, but its oral bioavailability is limited by its poor solubility. In this study, glycyrrhizic acid-baicalein nano-micelles were prepared by an ultrasonic-film hydration method. The baicalein-loaded nano-micelles were evaluated by encapsulation efficiency, baicalein loading, particle size, polydispersity index and ζ-potential. A Box-Behnken statistical design was applied to obtain the optimal formulation (glycyrrhizic acid: 90 mg, baicalein: 8 mg, water bath shaking time: 12 h, ultrasonication time: 10 min). Nano-micelles prepared with the optimal formulation improved the solubility of baicalein in water by more than 4500 times and were characterized by differential scanning calorimetry and Fourier-transform infrared spectroscopy. An in vitro drug release study determined the cumulative drug release of baicalein in pH 6.8 and pH 8.3 buffer medium, after 6 h to be 18.20% and 47.96%, respectively, which indicates that the nano-micelles have a sustained-release effect on baicalein and that the release rate can be modulated by changing the pH.

A comparative analysis of raw and processed Radix Paeoniae Alba samples using HPLC, chemometrics, and antioxidant activity.

Radix Paeoniae Alba (RPA) and its processed products are commonly used in traditional Chinese medicine, but the chaos phenomenon among processed products often occurs. In this study, we used multiple chemometric methods to analyze raw and six different processed products of RPA based on HPLC fingerprinting. Heat map analysis was used to assess the changes in chemical composition. Principal component analysis was used for classification, and the samples were divided into four classes: class 1 (raw, wine-processed, and vinegar-processed products), class 2 (bran-processed and soil-processed products), class 3 (stir-fried products), and class 4 (coke products). Further, the orthogonal partial least squares discriminant analysis model was used to obtain chemical markers among different classes. The antioxidant property of RPA is an important factor responsible for its pharmacological effects, and so the antioxidant activity of RPA was also investigated. We measured 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydroxyl radical scavenging ability, and ferric reducing antioxidant power. The total antioxidant activity follows the order: coke > stir-fried > soil-processed > bran-processed > wine-processed > raw > vinegar-processed products. These results suggest that different processing methods affect the chemical composition and antioxidant power of RPA, and thus, different products of RPA should not be mixed.

In vitro and in vivo evaluation of the influences of polysaccharides derived from Glycyrrhiza uralensis on three alkaloids and potential interaction mechanisms.

The purpose of this study was to investigate the influences of the polysaccharides derived from Glycyrrhiza uralensis Fisch. (GCPs) on aconitine (AC), hypaconitine (HA), and benzoylmesaconine (BMA) from Aconitum carmichaelii Debx. and to explore potential interaction mechanisms. Biopharmaceutical properties in vitro including stability, aqueous solubility and permeability were determined by UPLC. Pharmacokinetic parameters in vivo were determined using UPLC-MS/MS. Phase solubility analysis, UV-vis spectrophotometry and differential scanning calorimetry (DSC) were used to explore potential interaction mechanisms. The results showed that GCPs could increase the stabilities of three alkaloids and solubilities of AC and HA, significantly decrease permeabilities of three alkaloids. The pharmacokinetic studies demonstrated that, after combination with GCPs, AC exhibited a higher Cmax value, shorter t0.5, higher elimination rate and greater area under the concentration-time curve (AUC) value compared to free AC. GCPs also improved the Cmax, t0.5, AUC(0-tn) and AUC(0-∞) values of HA to play a therapeutic role, and improved the t0.5 and AUC(0-∞) values of BMA to prolong the pharmacological effect and increase bioavailability. In addition, the results for the apparent formation constants and DSC analysis showed that an inclusion complex could be formed between GCPs and AC, GCPs and HA, and GCPs and BMA.

Effects of Astragalus polysaccharide on the solubility and stability of 15 flavonoids.

Poor solubility and stability are limitation factors of flavonoids application. Polysaccharides from herbs have been proven to be functional foods and potential promising natural supplements with large molecular weight and complex structures. Astragalus polysaccharides (APS) are the main components of Astragalus membranaceus (Fisch.) and novel synergistic pharmacological effects between Astragalus polysaccharides and flavonoids have been reported. However, the general effects of polysaccharides when co-administered with flavonoids are currently unknown. Herein, the influences of APS on aqueous solubilities and stabilities of fifteen flavonoids were systematically investigated, and the mechanism of effects of polysaccharides on flavonoids was further considered. The results showed that APS could significantly enhance the solubilities and stabilities of the flavonoids, with solubilization effect improved by 68.88-fold for quercetin and negatively correlated with the aqueous solubility of the flavonoid itself. A phase solubility study and Differential scanning calorimetry characterization indicated that APS could form complexes with flavonoids at 1:1 ratio with K values ranging from 1491 to 55,395 L·mol-1, a tendency to improved solubilization at higher association constant values was also observed. Those findings could provide the basis for a new approach to solving the problems of poor solubility and stability of flavonoids through the application of natural macromolecules.

Comparative Investigation for Rotten Xylem (kuqin) and Strip Types (tiaoqin) of Scutellaria baicalensis Georgi Based on Fingerprinting and Chemical Pattern Recognition.

Scutellaria baicalensis Georgi (SBG) is not just as a traditional herbal medicine but also a popular functional food in China and other Asian countries. A sensitive simple strategy was developed for the first time to analyze SBG from eight different geographical sources using high-performance liquid chromatography (HPLC) coupled with multivariate chemometric methods. Two unsupervised pattern recognition models, hierarchical cluster analysis (HCA) and principal components analysis (PCA), and a supervised pattern recognition model, partial least squares discriminant analysis (PLS-DA), were used to analyze the chemical compositions and physical traits of SBG. The important chemical markers baicalin, baicalein, and wogonoside were analyzed quantitatively and with PLS-DA. These methods distinguished rotten xylem (kuqin) and strip types (tiaoqin) of SBG and found that the thickness of the slice had a significant impact on the classification of SBG. Two classes of strip types were identified: one as the uncut pharmaceutical, which was sectioned with a thickness >3 mm; the other as a thin-sectioned strip type, with a thickness of <2 mm. This fingerprinting technique coupled to a chemometric analysis was used for the simultaneous quantitation of three components (chemical markers) of SBG, and greatly simplified the complicated identification of the multiple components of this plant relative to traditional methods. The strategy can clearly distinguish between kuqin and tiaoqin of SBG, and suggests that the thickness of the slice can be used as the basis for evaluation of SBG. These data provide a theoretical basis and scientific evidence for the development and utilization of SBG.

Comparative investigation for raw and processed Aconiti Lateralis Radix using chemical UPLC-MS profiling and multivariate classification techniques.

A strategy combining chemical UPLC-MS profiling and multivariate classification techniques has been used for the comparison of raw and processed Aconiti Lateralis Radix. UPLC-MS was used to identify 18 characteristic compounds, which were selected for discrimination of the raw and two processed products (Heishunpian and Baifupian). Chemometric analyses, including the combination of a heat map and hierarchical cluster analysis (HCA) and principal component analysis (PCA), were used to visualize the discrimination of raw and two processed products. HCA and PCA provided a clear discrimination of raw Aconiti Lateralis Radix, Heishunpian and Baifupian. Finally, the counter-propagation artificial neural network (CP-ANN) was applied to confirm the results of HCA, PCA and to explore the effect of 18 compounds on samples differentiation and the rationality of processing. The results showed that this strategy could be successfully used for comparison of raw and two processed products of Aconiti Lateralis Radix, which could be used as a general procedure to compare herbal medicines and related processed products to elaborate the rationality of processing from the perspective of chemical composition.

Quality Evaluation of Phellodendri Chinensis Cortex by Fingerprint⁻Chemical Pattern Recognition.

Phellodendri Chinensis Cortex (PCC) and Phellodendri Amurensis Cortex (PAC) are increasingly being used as traditional herbal medicines, but they are often mistaken for each other. In this study, the fingerprints of PCC from six different geographical sources were obtained by high-performance liquid chromatography, and multivariate chemometric methods were used for comprehensive analysis. Two unsupervised pattern recognition models (principal component analysis and hierarchical cluster analysis) and a supervised pattern recognition model (partial least squares discriminant analysis) were established on the basis of the chemical composition and physical traits of PCC and PAC. PCC and PAC were found to be distinguishable by these methods. The PCC category was divisible into two categories, one with more crude cork and a maximum thickness of ~1.5 mm, and the other with less net crude cork and a maximum thickness of 0.5 mm. According to the model established by partial least squares discriminant analysis (PLS-DA), the important chemical marker berberine hydrochloride was obtained and analyzed quantitatively. From these results combined with chemometric and content analyses, the preliminary classification standards for phellodendron were established as three grades: superior, first-order and mixed. Compared with the traditional identification methods of thin layer chromatography identification and microscopic identification, our method for quality evaluation is relatively simple. It provides a basis and reference for identification of PCC and enables establishment of grade standards. It also could be applied in quality control for compound preparations containing PCC.

The influence of polysaccharides from Ophiopogon japonicus on 2,3,5,4'-tetrahydroxy-stilbene-2-O-ß-d-glucoside about biopharmaceutical properties in vitro and pharmacokinetics in vivo.

2,3,5,4'-Tetrahydroxy-stilbene-2-O-ß-d-glucoside (THSG) is a bioactive polyhydroxystilbene compound obtained from Polygonum multiflorum Thunb. Although it has broad therapeutic potential, the application of THSG is limited due to its poor stability in vitro, poor absorption in the intestine, and short-term storage in tissues. Polysaccharides from herbal medicines have been proven to be functional foods and potential natural plant derived drugs. However, the effects of polysaccharides when co-administered with other active ingredients are currently unknown. The present study investigated the influences of Ophiopogon japonicus polysaccharides (OJPs) on THSG. Results showed that OJPs notably enhanced aqueous solubility, and stability of THSG, but slightly decreased the permeability of THSG. In addition, Tmax, Cmax, and AUC(0-tn) of THSG were 3.5 fold, 1.45 fold and 2.32 fold higher for THSG-OJP. Thus, OJPs could potentially be used to improve the biopharmaceutical properties and prolong the pharmacological effects of THSG. This finding could provide a reference point for further applications of polysaccharides from herbal medicines.

Development and Validation of an UPLC-MS/MS Method for Pharmacokinetic Comparison of Five Alkaloids from JinQi Jiangtang Tablets and Its Monarch Drug Coptidis Rhizoma.

JinQi Jiangtang (JQJT) tablets, a Chinese patent medicine approved by the State Food and Drug Administration, are composed of Coptidis Rhizoma, Astragali Radix, and Lonicerae Japonicae Flos, and have a significant effect on diabetes. Coptidis Rhizoma is monarch drug in the prescription. The aim of the present study was to investigate and compare the pharmacokinetics of multiple ingredients from JQJT tablets and Coptidis Rhizoma extract (CRE) following oral administration in rats. Five alkaloids: coptisine chloride, epiberberine chloride, berberine chloride, jatrorrhizine chloride, and palmatine chloride, were simultaneously determined in rat plasma using established and validated ultra-high performance liquid chromatography mass spectrometry (UPLC-MS/MS). Significant pharmacokinetic differences were observed for the five alkaloids after a single administration of CRE and JQJT tablets. Compared with CRE, the Cmax values of palmatine chloride and jatrorrhizine chloride were decreased significantly, the AUC0-t values of four alkaloids (all except jatrorrhizine chloride) were notably decreased, and the mean residence times of all five alkaloids were significantly decreased after administration of JQJT tablets. The results indicated that the absorption characteristics of the five alkaloids from Coptidis Rhizoma would be influenced by the compatibility of Astragali Radix or Lonicerae Japonicae Flos from JQJT tablets, such that absorption was inhibited and elimination was accelerated. In conclusion, the developed strategy was suitable for the comparison of five alkaloids from JinQi Jiangtang tablets and its monarch drug, which could be valuable for compatibility studies of traditional Chinese medicines.