Chromatographic Fingerprint Analysis Research Articles

Acute toxicity appraisal of Saussurea heteromalla extract and development of its chemotherapeutic herbal dosage form.

Ethnopharmacological relevance of Saussurea species for anti-cancer compounds instigated us to develop chemotherapeutic herbal tablets. This study was an ongoing part of our previous research based on the scientific evaluation of Saussurea heteromalla (S. heteromalla) for anti-cancer lead compounds. In the current study, S. heteromalla herbal tablets (500 /800 mg) were designed and evaluated for anti-cancer activity. Arctigenin was found as a bioactive lead molecule with anti-cancer potential for cervical cancer. The in vitro results on the HeLa cell line supported the ethnopharmacological relevance and traditional utilization of S. heteromalla and provided the scientific basis for the management of cervical cancer as proclaimed by traditional practitioners in China. LD50 of the crude extract was established trough oral acute toxicity profiling in mice, wherein the minimum lethal dose was noticed as higher than 1000 mg/kg body weight orally. Chromatographic fingerprint analysis ensured the identity and consistency of S. heteromalla in herbal tablets in terms of standardization of the herbal drug. About 99.15% of the drug (S. heteromalla crude extract) was recovered in herbal tablets (RSD: 0.45%). In vitro drug release profile was found to be more than 87% within 1 h, which was also correlated with different mathematical kinetic models of drug release (r2 = 0.992), indicating that drug release from matrix tablets into the blood is constant throughout the delivery. The dosage form was found stable after an accelerated stability parameters study which may be used for anti-cervical cancer therapy in the future, if it qualifies successful preclinical investigation parameters.

Systematic evaluation of Rutongshu oral liquid based on chromatographic fingerprinting and nontargeted metabolomic analysis

AbstractBased on chromatographic fingerprinting combined with quantitative analysis on characteristic chemical constituents as well as hierarchical cluster analysis, an easy and sensitive approach utilizing high performance liquid chromatography (HPLC) was developed for the identification and quality evaluation of Rutongshu oral liquid (ROL). What is more, nontargeted metabolomic analysis was conducted to gain a global view in terms of its chemical profile. In this study, 16 peaks from different batches (S1–S10) of ROL samples produced by Taihe Hospital of Chinese Medicine were selected as common peaks for the evaluation of their similarity whose values of all tested 10 batches exceeded 0.90 when compared with the control fingerprints. Meanwhile, simultaneous quantification of five markers in the oral solution, including albiflorin, paeoniflorin, chlorogenic acid, quercetin and ferulic acid was performed, and standard curves established for respective reference substances showed good regression in the linear range (r2 > 0.999) with recoveries in the range of 98.96–102.35%. The ultra-high performance liquid chromatography (UHPLC) combined with Orbitrap Exploris 120 mass spectrometer resulted in 88 identified compounds. The results of validation showed that the established method was reproducible, precise and stable. This study offers an effective, dependable and useful approach for the systematic evaluation of the hospital formulation ROL.

Quality evaluation of Lonicerae Japonicae Flos from different origins based on high-performance liquid chromatography (HPLC) fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition.

Lonicerae Japonicae Flos (LJF) is widely used in food and traditional Chinese medicine. To meet demand, Lonicera japonica Thunb. is widely cultivated in many provinces of China. However, reported studies on the quality evaluation of LJF only used a single or a few active components as indicators, which could not fully reflect the quality of LJF. In the present study, we aimed to develop a methodology for comprehensively evaluating the quality of LJF from different origins based on high-performance liquid chromatography (HPLC) fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition. The HPLC method was developed for fingerprint analysis and was used to determine the contents of 19 components of LJF. To distinguish between samples and identify differential components, similarity analysis, hierarchical cluster analysis, principal component analysis, and orthogonal partial least squares discriminant analysis were performed. The HPLC fingerprint was established. Using the developed method, the contents of 19 components recognized in the fingerprint analysis were determined. Samples from different origins could be effectively distinguished. HPLC fingerprinting and multicomponent quantitative analysis combined with chemical pattern recognition is an efficient method for evaluating LJF.

Discriminating the leaves of Ocimum sanctum Linn. conferring to their age by gas chromatographic fingerprint analysis assisted with multivariate analysis

Ocimum species is having massive variations in their chemical composition that includes day time, terrestrial and cyclical variation according to the season etc. In this study, variation in overall essential oil constituents among matured, intermediate and young leaf samples of O. sanctum was studied. The leaf samples according to their age- matured, intermediate and young leaves were collected from three different plants at 11 a.m. each. Extract of collected leaf samples were then scrutinized using gas chromatographic technique having flame ionization detector (GC-FID). Result reveals that the noteworthy variance was observed in overall essential oil constituents of the leaf samples according to their age. The Average chromatographic fingerprint data of triplicate run for different leaf samples were then treated to multivariate methods such as hierarchical cluster analysis and principal component analysis. In this study, instead of considering one or two biomarkers, we have exasperated to obtain as many constituents in chloroform extracts of chosen leaf samples. Scree plot reveals that nine components were responsible for the whole variation. Score plot and dendogram efficiently discriminated all the three leaf types according to their age in different clusters. The outcome as an inference was, O. sanctum leaves according to their age exhibit variation in its chemical composition and developed tool can be used for routine quality control.

Identification of Q-markers for Schisandrae Sphenantherae Fructus in treating drug-induced liver injury based on network pharmacology, fingerprint and quantitative analysis

This study aims to establish the ultra-performance liquid chromatography(UPLC) fingerprint and multi-indicator quantitative analysis method for Schisandrae Sphenantherae Fructus(SSF) and to screen out the potential quality markers(Q-markers) of hepatoprotection based on network pharmacology. The similarity analysis was performed using the Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System, which showed that the similarity of the fingerprints of 15 samples from different regions ranged from 0.981 to 0.998. Eighteen common components were identified, from which 3 differential components were selected by cluster analysis and principal component analysis. The "component-target-pathway" network was built to predict the core components related to the hepatoprotective effects. Fourteen core components were screened by network pharmacology. They acted on the targets such as AKT1, CCND1, CYP1A1, CYP3A4, MAPK1, MAPK3, NOS2, NQO1, and PTGS2 to regulate the signaling pathways of lipid metabolism and atherosclerosis, hepatitis B, interleukin-17, and tumor necrosis factor. Considering the chemical measurability, characteristics, and validity, schisantherin A, anwulignan, and schisandrin A were identified as the Q-markers. The content of schisantherin A, anwulignan, and schisandrin A in the test samples were 0.20%-0.57%, 0.13%-0.33%, and 0.42%-0.70%, respectively. Combining the fingerprint, network pharmacology, and content determination, this study predicted that schisantherin A, anwulignan, and schisandrin A were the Q-markers for the hepatoprotective effect of SSF. The results can provide reference for improving the quality evaluation standard and exploring the hepatoprotective mechanism of SSF.

Investigation on ethanolic stem extract of Ipomea sagittifolia to explore the presence of Saponins by High Performance Thin Layer Chromatography

Background: Saponins are a diverse group of naturally occurring plant compounds that play important roles in various aspects of plant physiology and ecology. An extract composed of several phytoconstituents may be easily analysed and interpreted using high performance thin layer chromatography (HPTLC) fingerprint analysis from a qualitative standpoint. Ipomea sagittifolia (Burn. f.) has a long history of use in traditional medical systems. Method: It was decided to interpret the phytoconstituents, namely the steroids in Ipomea sagittifolia (Burn. f.), in line with Current Good Manufacturing Practises, which highlight the significance of quality in relation to phytoconstituents, because HPTLC is a very trustworthy method of analysis. In this work, an HPTLC finger print profile for the saponins was made using an ethanolic extract of the stem of Ipomea sagittifolia (Burn. f.). This system includes the Scanner 4, TLC Visualizer, and Linomat 5 Applicator. Results: When phytoconstituents were evaluated using HPTLC densitometric screening at wavelengths of 366 nm and 540 nm, several peaks could be seen in the chromatogram. By analysing the peak regions, peak heights, and Rf values that were given in the proper tables, the phytochemicals are appraised. Conclusion: The study revealed the presence of saponins. This information is highly useful in studying chemical profiling and discovering bioactive components when the Rf values of these chemicals are compared with standards as a reference.

STANDARDISATION OF MURUNGAI NEI: A SIDDHA FORMULATION

Murungai Nei (Moringa ghee) is a multi-herbal formulation indicated as an aphrodisiac and spermatogenic in Siddha literature. The study aims to standardise the Murungai Nei’s quality, purity, and safety through its physicochemical, microbiological, chromatographic, and biochemical parameters. Physicochemical parameters like loss on drying, total ash, acid-insoluble ash, water-soluble ash, water-soluble extractive, alcohol-soluble extractive, and pH were found to be 0.03%, 0.0499%, 0.0199%, 0.07%, 0.1934%, and 18.49% respectively, which denote the purity and quality of the drug in this study. Qualitative preliminary phytochemical analysis reveals the presence of major phytochemicals: alkaloids, carbohydrates, saponin, phenols, tannins, flavonoids, diterpenes, quinones, gum, and mucilage. High-performance thin layer chromatography (HPTLC) fingerprinting analysis of the test drug reveals the presence of seven eminent modes relevant to the presence of seven different phytocomponents within it, whose Rf values range from 0.03 to 0.24. Biochemical analysis reports the presence of carbonate, sulphides, phosphate, ferrous, and magnesium. The results obtained from all the analyses of this study provide data on the pharmacogenetic (physical, chemical, and biochemical) properties of the Siddha poly-herbal formulation Murungai Nei. In the future, these data can be utilised as references for standardising the drug Murungai Nei.

A novel process for developing a quantitative chromatographic fingerprint analysis method based on analytical quality by design: A case study of Xiaochaihu capsules

In this study, an improved Analytical Quality by Design (AQbD) process was proposed to establish quantitative fingerprints of Chinese medicines. A quantitative fingerprint method for the strong polar components in Xiaochaihu capsules was developed as an example. Different method evaluation indicators were used to characterize this method, and these indicators were measured at different stages of quantitative fingerprint development. Through this method, the difficulty in determining common peaks in the early stage of quantitative fingerprint development is eliminated. The method parameters that needed to be optimized were the mobile phase gradient conditions, column temperature and flow rate through the Ishikawa diagram. Single-factor experiments were used to determine some parameters. Definitive screening design (DSD) experiments were used to optimize critical method parameters (CMPs). The common peaks were determined, and indices were optimized. The critical method attributes (CMAs) were the resolution of guanosine, the percentage of common peak to the total peak area, retention time of the last peak and the common peak number. A stepwise regression method was used to build quantitative models for critical method attributes and critical method parameters. The method operable design region (MODR) was calculated by the experimental error simulation method and successfully verified. After the analysis conditions were optimized, four components, uridine, adenine, 5-hydroxymethylfurfural, and guanosine, were quantitatively determined. There were 7 common peaks in the fingerprints. The common peak area accounted for 90.64%. Both fingerprint and quantitative detection analysis methods were methodologically validated. The new AQbD process proposed in this study can be potentially applied to develop other Chinese medicines fingerprint methods.

Simultaneous identification and quantification of pentacyclic triterpenoids and phenolic compounds from the leaves of Boswellia serrata using LC-MS/MS tandem mass spectrometry.

Boswellia serrata (B. serrata) is an important medicinal plant widely used as dietary supplements to provide a support for osteoarthritic and inflammatory diseases. The occurrence of triterpenes in leaves of B. serrata is very little or none. Therefore, the qualitative and quantitative determination of phytoconstituents (triterpenes and phenolics) present in the leaves of B. serrata is very much needed. The aim of this study was to develop an easy, rapid, efficient and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method for the identification and quantification of the compounds present in the leaves extract of B. serrata. The purification of ethyl acetate extracts of B. serrata was performed by solid phase extraction method, followed by HPLC-ESI-MS/MS analysis. Chromatographic parameters of the analytical method included negative electrospray ionization (ESI-) with a flow of 0.5mL/min in gradient mode consisting of acetonitrile (A) and water (B) containing 0.1% formic acid, at 20°C. Total 19 compounds (13 triterpenes and 6 phenolic compounds) were separated, and simultaneously quantified using a validated LC-MS/MS method with high accuracy and sensitivity. Good linearity was obtained with r2 > 0.973 in the calibration range. The overall recoveries were in a range between 95.78 and 100.2% with relative standard deviations (RSD) below 5% for the entire procedure of matrix spiking experiments. Overall, there was no ion suppression from the matrix. The quantification data showed that the total amount of triterpenes and phenolic compounds in the leaves of B. serrataethyl acetate extract samples ranged from 14.54 to 102.14mg/g and 2.14 to 93.12mg/g of dry extract, respectively. This work provides, for the first time, a chromatographic fingerprinting analysis on the leaves of B. serrata. A rapid, efficient, and simultaneous liquid chromatography-mass spectrometry (LC-MS/MS) method was developed and used for the both identification and quantification of triterpenes and phenolic compounds in the leaves extracts of B. serrata. The method established in this work can be used as quality-control method for other market formulations or dietary supplements containing leaf extract of B. serrata.

Identification of Antibacterial Components in Indonesian Minor Spices Using Chromatographic Fingerprinting and Multivariate Data Analysis

ABSTRACT This study determined the antibacterial activity of 12 minor Zingiberaceae spices and to identify the active compounds in the most active spices using chromatographic fingerprinting and multivariate data analysis. Curcuma purpurascens had the highest antibacterial activity against Staphylococcus aureus and Escherichia coli. HPLC chromatogram and antibacterial data of hexane, chloroform, ethyl acetate, methanol, and water extracts of C. purpurascens were linked using Orthogonal Projection to Latent Structure. Retention time of 16.5–18.0 min correlated with antibacterial activity and is abundantly found in methanol fraction. UHPLC-HRMS analysis of methanol fraction showed that the peak was attributed to demethoxycurcumin.

Analysis on quality value transmitting of Qinggusan reference sample of classical prescription based on quantitative fingerprint technology

Qinggusan is the 69th prescription in the first batch of "Catalogue of Ancient Chinese Classic Formulas". In modern clinical practice， Qinggusan is mainly used to treat noninfectious fever. However， because few studies on Qinggusan reference samples and their quality value transfer are available， the development and promotion of its compound preparations are restricted. Therefore， establishing an accurate and comprehensive quality control method to clarify the critical quality attributes of Qinggusan reference samples is of great importance. In this study， 15 batches of Qinggusan reference samples were processed to determine the range of their dry extract ratios. Quantitative high-performance liquid chromatography （HPLC） fingerprint analysis was performed using a Waters Symmetry Shield RP18 column （250 mm×4.6 mm， 5 μm） with acetonitrile-0.1% （v/v） formic acid aqueous solution as the mobile phase in gradient elution mode. The flow rate was 1.0 mL/min， the column temperature was 30 ℃， and the detection wavelength was 254 nm. The HPLC fingerprints of the Qinggusan reference samples were established under these conditions to evaluate their similarity. The established method was systematically validated and found to demonstrate good precision， repeatability， and sample stability. Subsequently， characteristic peaks were identified and attributed by HPLC-quadrupole-time-of-flight-mass spectrometry （HPLC-Q-TOF-MS） analysis. MS was performed in electrospray ionization mode， the data were collected in both positive- and negative-ion modes， and the detection range was m/z 50-2000. The contents and transfer rate ranges of the index components， namely， gentiopicrin， mangiferin， picroside Ⅱ， picroside Ⅰ， and glycyrrhizic acid， were determined to analyze the quality value transfer of the samples. The results demonstrated that the dry extract rate of the 15 batches of Qinggusan reference samples ranged from 24.10% to 26.88% and that their fingerprint similarities were generally greater than 0.95. Twelve common peaks were identified by reference identification， literature comparison， and high-resolution MS analysis. Twelve compounds， including six iridoid glycosides， two flavonoids， one alkaloid， one triterpenoid saponin， and two others. Among them， L-picein， androsin， picroside Ⅳ， picroside Ⅱ and picroside Ⅰ were from Picrorhizae Rhizoma， loganin acid， swertiamarin and gentiopicrin were from Gentianae Macrophyllae Radix， neomangiferin and mangiferin were from Anemarrhenae Rhizoma， dichotomine B was from Stellariae Radix， and glycyrrhizic acid was from Glycyrrhizae Radix et Rhizoma. The five key components presented good linear relationships in their respective linear ranges， and all correlation coefficients were higher than 0.999. The relative standard deviations （RSDs） of precision， stability， and repeatability were less than 1.3%. The average recoveries varied between 95.92% and 102.5%， with RSDs less than 3.9%； these values meet the requirements defined in the 2020 edition of the Chinese Pharmacopoeia. The contents of gentiopicrin， mangiferin， picroside Ⅱ， picroside Ⅰ， and glycyrrhizic acid in the 15 batches of reference samples were in the range of 17.92-27.55， 1.83-4.42， 23.08-36.44， 8.43-15.04， and 0.94-2.39 mg/g， respectively， and their transfer rates from the decoction pieces to the reference samples were 47.91%-63.95%， 22.96%-59.39%， 60.82%-77.82%， 64.25%-99.53%， and 15.30%-39.30%， respectively. In this study， the chemical components of Qinggusan reference samples were comprehensively identified and their quality value transfer was studied through the combination of HPLC fingerprinting and MS. Clarification of the critical quality attributes of Qinggusan reference samples could provide a basis for the quality control of Qinggusan compound preparations.

Identification of <i>Sida rhombifolia</i> from Its Related Plants Using Thin-Layer Chromatographic Analysis

Sida rhombifolia belongs to the Malvaceae family and is often used to treat gout in Indonesia. S. rhombifolia has many efficacies and contains many different chemical components. The abundance and variation of chemical content and chemical compounds in this medicinal plant are challenging factors in ensuring medicinal plants' safety and quality control. Thin-layer chromatography (TLC) fingerprint analysis derived from S. rhombifolia extract can also be used for the medicinal plant's quality control. This research aimed to develop the optimum condition for the chemical fingerprint analysis of S. rhombifolia using a TLC fingerprint analysis. A total of 11 bands were produced with optimum separation using silica gel 60 F254 TLC plate, a mixed mobile phase condition with chloroform, ethyl acetate, and methanol (6.5:2:1.5). This fingerprint analysis performed an excellent separation in the TLC plate at 366 nm with sulfuric acid as reagent derivatization. In general, the results of the analysis validation, including stability, specificity, precision, and robustness of TLC fingerprints, met the acceptance criteria. The TLC fingerprint of S. rhombifolia can be distinguished from 2 related plants with similar leaf shapes, Turnera ulmifolia L. and Hibiscus rosa-sinensis. The developed method was validated, so it could be used to control S. rhombifolia quality.

Liquid Chromatography Fingerprint Analysis of Released Compounds in Plasma Samples of Stroke Patients after Thrombolytic Treatment

Plasma samples obtained from stroke patients treated with recombinant tissue-type plasminogen activator (rt-PA) and not treated with rt-PA were evaluated with different HPLC methodologies to obtain information about the possible release of small molecules as a result of the thrombolytic treatment. Plasma samples, without derivatization and derivatized with 6-aminoquinolyl-N-hydroxysuccinimidyl carbamate (AQC), were evaluated with a HPLC gradient method, which consisted of a mobile phase of 10 mM ammonium acetate buffered solution (pH = 5.3) and acetonitrile. Three different detection methods were applied: UV, fluorescence, and ESI-MS. The results obtained showed that a group of new highly hydrophilic compounds appeared in most samples analyzed from treated patients, just after the administration of rt-PA. These compounds appeared shortly after the administration of the drug and were detected during the first 24 h after treatment, disappearing from plasma after this time. These new compounds were not detected either in controls or in non-treated stroke patients, which suggests that they were released into the plasma as a consequence of the thrombolytic effect of the drug. Our results suggest that these new compounds might be free glycans. The use of AQC as a derivatizing reagent has demonstrated that the new compounds detected cannot contain primary or secondary amine groups in their structure. The molecular mass determined by ESI-MS (821 Da) suggests that if these compounds are free glycans they might be a high-mannose type.

Comparative bioactivity evaluation of secondary metabolites from the leaves, stem and flowers of Tithonia diversifolia

Natural products have been extensively described as important in the process of drug.discovery and development. Drugs based on natural products are used in the treatment of infectious diseases, cancer, lipid disorders and immunomodulation. These drugs illustrate the importance of natural products as sources of innovative therapeutic agents. Recent studies point towards therapeutic potential of large number of natural products that originate from microbes and plants. These type of natural products plays a crucial role to shape the future of drug discovery process. Numerous studies indicate the use of Tithonia diversifolia (TD) as folk medicine to cure diseases like malaria, jaundice, hepatitis and dermatological problems. The present study is focused on identifying and comparing the bioactivity of different extracts from TD. Bioactive extracts of the leaves, stem, flowers were subjected to quantitative and qualitative phytochemical analysis. Qualitative analysis of TD extracts showed the presence of terpenoids, flavanoids, alkaloids, phenolics as the major class of compounds. High-Performance Thin-Layer Chromatography (HPTLC) fingerprinting analysis was carried to establish a quantitative profiling of the extracts. GC–MS of the extracts indicated the presence of Hexadecanoic acid, 2,3- dihydrobenzofuran, Squalene, dotriacontane, 1- hexacosanol, D- tagatose as the major phytocompounds present in the TD Hexane and ethyl acetate extract. The tested Tithonia diversifolia extracts showed significant anti-oxidant and anti-bacterial activities indicating therapeutic potential for TD.

High-performance Thin-layer Chromatography Fingerprinting Analysis of Punarnavadi Kwatha

Introduction: Punarnavadi kwatha is one among the commonly used medicine in Ayurvedic practice, mentioned in Chakradatta in sopha chikitsa. Main indications of Punarnavadi kwatha are Sarvanga sopha (generalised oedema), Udara (ascites), Kasa (cough), Soolam (colicky pain), and Swasa (dyspnoea) associated with Pandu (anaemia). For the purpose of quality assurance and uniformity, scientific verification of Ayurvedic medications is important. Usage of modern analytical techniques helps in the proper authentication of medicines. In the present study, highperformance thin-layer chromatography (HPTLC) fingerprinting of Punarnavadi kwatha aimed to assess the different phytoconstituents. Materials and Methods: Methanolic extract of Punarnavadi kwatha was subjected to HPTLC fingerprinting using the solvent system toluene:ethyl acetate:formic acid (6:3:0.5). Toluene is a least polar solvent, ethyl acetate is mid polar solvent and formic acid is highly polar so in this solvent system, maximum separation of compounds took place. Results and Discussion: The number of peaks obtained at visualization 254 nm was 8 and total area of compounds in is 30309.5. Visualization at 366 nm, number of peaks was 12 and total area is 42473.2. Conclusion: Thus, the study revealed that HPTLC fingerprinting of formulations can be used in the quality control of ayurvedic medicaments. The maximum separation of compounds occurred at this solvent system.

Antioxidant Capacity, Total Phenolic, and Flavonoid Content from Java Tea (Orthosiphon aristatus) Extracts

Orthosiphon aristatus, known as kumis kucing in Indonesia, has some biological activity, i.e., as an anti-inflammatory, fever, rheumatism, diabetes, and kidney stone. This plant is also known for being rich in phenolic compounds such as flavonoids. In this study, we would like to compare the antioxidant activity, total phenolics, and flavonoid content from the stems and leaves of Orthosiphon aristatus extracted with water and ethanol by maceration. The yield stem and leaves of the water extract are 12.98-31.28%, and ethanol extract 3.58 - 9.82% (dry basis). The stem and leaves water extract contains a phenolic of 7.82-26.43 (mg GAE/g) and ethanol extract of 2.27-7.84 (mg GAE/g). Water extract Orthosiphon aristatus flavonoid content is 1.23-2.86 (mg EK/g), and ethanol extract is 1.09-3.95 (mg EK/g). The antioxidant activity of ethanol (IC50), extract from leaves was 123.91 (µg/mL) and from stems was 125.50 (µg/mL), while IC50 of water extract from leaves was 114.70 (µg/mL) and from stems was 125.03 (µg/mL). The correlation test result of active compounds using linear regression is phenolic compounds with antioxidants stronger than flavonoids. The qualitative thin-layer chromatography fingerprint analysis showed that ethanol and water extract contained natural sinensetin compounds.

Development of a Quantitative Chromatographic Fingerprint Analysis Method for Sugar Components of Xiaochaihu Capsules Based on Quality by Design Concept

Background: Xiaochaihu capsule is composed of seven traditional Chinese medicines. The pharmacopoeia only focuses on the quantitative detection of baicalin, which cannot fully reflect the quality of the preparation. Some medium polar components were used to establish the fingerprint of Xiaochaihu capsule, but there was no report on the strong polar components. Methods: A high performance liquid chromatography-corona charged aerosol detection technology was used to establish a fingerprint analysis method for Xiaochaihu capsules following an analytical quality by design approach. Definitive screening designed experiments were used to optimize the method parameters. A stepwise regression method was used to build quantitative models. The method operable design region was calculated using the experimental error simulation method. Plackett–Burman designed experiments were carried out to test robustness. Results: The contents of four components were simultaneously determined. There were seven common peaks in the fingerprint. The common peak area accounted for 91.72%. Both fingerprint and quantitative analysis methods were validated as applicable in the methodology study. The quantitative fingerprint analysis method for sugar components can fill the gap in the detection of strong polar components in the existing methods. It provides a new technology for the comprehensive overall evaluation of Xiaochaihu capsule.

In vitro regeneration and chromatographic fingerprint analysis of Aphanamixis polystachya (Wall.) Parker by HPTLC technique

• In vitro regeneration on nodes of Aphanamixis polystachya tree showed breakage of recalcitrance. • Biomass content was higher on cytokinins 2-ip (N 6 -(2-Isopentyl) Adenine) supplemented Murashige and Skoog's medium. • HPTLC analysis of showed higher diversity of phytochemicals in the in vitro shoots without any supplementation of elicitors. • Comparison between the in vitro shoots and mature plants shown the presence of majority of phytochemicals in both groups. Aphanamixis polystachya is an evergreen tree belonging to the Meliaceae family used for centuries in traditional medicines. Various metabolites that were isolated from the barks showed high therapeutic value. Isolation of pharmaceutically active compounds imposes pressure on harvesting the plants from the wild, resulting in the endangerment of the tree. The development of an alternate method for the isolation of phytochemicals is the need of the hour. The current study evaluated the phytochemical screening of bioactive compounds from the tree parts and in vitro regenerated shoots of A. polystachya . Due to its recalcitrancy, tissue culture data remains inaccessible, and the present investigation was first attempt on developing a protocol for generating plantlets using in vitro culture. Murashige and Skoog's medium fortified with cytokinins like BAP (6-Benzylaminopurine), Kinetin, 2-ip (N 6 -(2-Isopentyl) Adenine), and TDZ (Thidiazuron) were used, and maximum biomass was produced from 3 mg/L of 2-ip. Meanwhile, HPTLC fingerprinting analysis on the tree parts (bark and leaves) and in vitro shoots showed similar diversity among metabolites. The secondary metabolites produced in the in vitro plants were the same with mature trees for standards like Rutin, Stigmasterol, β-Sitosterol, Piscidinol A, Bourjotinolone B, and Niloticin.

Combining network pharmacology with chromatographic fingerprinting and multicomponent quantitative analysis for the quality evaluation of Moutan Cortex.

In this study, we combined network pharmacology, chromatographic fingerprinting and multicomponent quantitative analysis to evaluate the quality of Moutan Cortex (MC). Specifically, through network pharmacology, we obtained a comprehensive understanding of the active components and pharmacological activities of MC. In addition, the method of establishing fingerprint and multicomponent quantification by ultra high-performance liquid chromatography is convenient and comprehensive, and can more fully reflect the overall distribution of various chemical components. Principal component analysis and discriminant least square analysis were used. The results show that MC plays a synergistic role through multiple targets and pathways. The contents of chemical components in MC from different sources were significantly different. Combining network pharmacology and multicomponent quantitative results, gallic acid, benzoyl paeonol, paeonol, methyl gallate, benzoic acid and paeonol can be used as quality markers for MC quality control. This study provides a comprehensive and reliable strategy for the quality evaluation of MC and determines its quality indicators to ensure the quality of Chinese herbal medicine.

Phytochemical studies and HPTLC fingerprint profiling of different extracts of Portulaca Quadrifida L.

The present study was designed to detect the class of compound by phytochemical evaluation and perform the high performance thin layer chromatography (HPTLC) fingerprint analysis of hexane extract of P. quadrifida (HEPQ), chloroform extract of P. quadrifida (CEPQ), ethyl acetate extract of P. quadrifida (EAEPQ) and ethanol extract of P. quadrifida (EEPQ). The successive extraction based on eluotropic solvent system was utilised for extraction. The preliminary phytochemical screening was carried out for all the extract. For the HPTLC fingerprint profiling the precoated Merck silica plates were used as stationary phase while, mobile phase utilised was Toluene: Chloroform: Ethanol (5:4:1 v/v/v). The scanning was done at White light, 254 nm and 366 nm. The presence of steroids, coumarins, flavonoids, glycosides, saponins, coumarins was found in all 4 extracts, except the terpenoids which were present in HEPQ and EAEPQ. The fingerprinting profile was successfully obtained for all four extracts of P. quadrifida. This profiling data can thus be utilized for quality control of the P. quadrifida both on qualitative and quantitative basis.