High-performance Liquid Chromatography Chromatograms Research Articles

Chromatographic authentication of botanical origin: Herbaceous pollen profiling with HPLC, HPTLC and GC-MS analysis.

This study describes a robust chromatographic authentication methodology for herbaceous pollen, employing gas chromatography-mass spectrometry (GC-MS), high-performance liquid chromatography (HPLC) and high-performance thin liquid chromatography (HPTLC) protocols. The comprehensive profiling of organic compounds not only distinguishes between different botanical sources but also establishes a reliable framework for quality control and assessment of herbaceous pollen authenticity. Traces of quercetin were detectable using HPTLC in Chaenomeles japonica, and the composition of the mobile phase led to distinct phenolic acid tracks in the extracts of free phenolic compounds. In Lonicera nummulariifolia, prominent chlorogenic acid signal and traces of 3,4-dihydroxybenzoic acid were identified, along with the presence of vanillic, trans-ferulic, p-coumaric and p-hydroxybenzoic and sinapic as phenolic acid standards. The HPLC chromatogram identified six peaks representing bioactive phenolic compounds such as gallic acid measuring 5.89 ± 0.56 mg g-1 , hydroxybenzoic acid 2.39 ± 0.78 mg g-1 and caffeic acid 2.83 ± 0.11 mg g-1 . The combined use of GC-MS, HPTLC and HPLC techniques provides a powerful and reliable means of authenticating the botanical origin of herbaceous pollen, offering valuable insights for quality control and ensuring the accuracy of botanical source identification.

Effect of Fertilization on Phenolics of Rapeseeds and Their Antioxidant Potential.

Three varieties of rapeseed (Castilla, California, and Nelson F1) were cultivated using medium-intensive (control), intensive, and economical (spare) technologies with different nitrogen and sulfur fertilization techniques. The antioxidant potential of rapeseeds was investigated using ABTS, FRAP, and DPPH assays. The content of total phenolic compounds was determined using the Folin-Ciocalteu phenol reagent. The profile of phenolic compounds was determined using high-performance liquid chromatography (HPLC). Diversifying fertilization in various ways influenced the content of phenolic compounds in extracts of rapeseed. In extracts from the Nelson F1 rapeseeds, intensive cultivation resulted in a lower content of phenolic compounds compared to the control group. Economic fertilization reduced the content of phenolic compounds in seeds from the California variety. HPLC chromatograms of the extracts were characterized by the presence of five (California and Castilla) and six (Nelson F1) main phenolic compounds. Two compounds were identified as sinapine and sinapic acid; others were classified as derivatives of sinapic acid. The effect of fertilization on the antioxidant activity of the seeds and their extracts varied depending on the plant variety and antioxidant assay. For the Castilla and California varieties, no differences were found in the results of the ABTS assay. The antiradical activity against ABTS•+ of extracts from the Nelson F1 intensive and spare cultivated seeds was higher than that of extracts from control seeds. The FRAP values of extracts/seeds from the Castilla variety cultivated using different methods did not differ significantly. The results of the DPPH assay were not affected by fertilization in the case of extracts from the California and Castilla varieties. However, the extracts from spare cultivated seeds of Nelson F1 exhibited stronger antiradical activity against DPPH•. These findings highlight the complex relationship between fertilization practices, phenolic compound accumulation, and antioxidant activity in rapeseed. Integrating varietal traits and cultivation practices is crucial for optimizing the nutritional benefits of rapeseed.

Fermentation medium optimization of Streptomyces sp. as an antifungal agent against the Ganoderma boninensis pathogen in oil palm

Ganoderma boninensis is the most common fungus which attacks oil palm trees. However, a significant percentage of inhibition to the problem is found through the use of Streptomyces sp. The optimization of the Streptomyces sp. fermentation medium growth factors affects the secondary metabolites production. This study aimed to identify the best formulation of carbon and nitrogen sources and the optimum concentration of Streptomyces sp. fermentation medium for antifungal compound production. The results showed that the best sources of carbon and nitrogen were liquid glucose and monosodium glutamate in the inhibition zones of 16.7 mm and 6.3 mm, while the best concentration levels were 20 g/L and 14.19 g/L, respectively. The results of the first optimization showed an inhibition zone response and area (%) of the optimum high‐performance liquid chromatography (HPLC) chromatogram of 24.39 mm and 62.68 percent, respectively. Taking the suggestion of the first optimization, the second optimization produced 15.2 g/L and 8.3 g/L. The predicted value of the inhibition zone was 21.47 mm, and the area (%) of the HPLC chromatogram was 53.44 percent. The validation results showed an inhibition zone response of 22.01 mm and an HPLC chromatogram area (%) of 54.86 percent. The difference between the predicted and validation values was less than 5 percent; the validation value was thus in line with the value predicted by Design Expert 10.0.7. The chemical formula of the probable active compound is that of the cyclo(phenylalanyl‐prolyl) compound.

BIOCHEMICAL EVALUATION OF INDUS VIVA I PULSE NATURAL AYURVEDIC SYRUP AND IT’S IN SILICO – INTERACTION ANALYSIS

Objectives: In developing countries like India, diabetes mellitus and human bone cancer are progressively increasing which is global encumbrance. The objective of the study is to find out the phytoconstituents of Indus Viva I Pulse health drink extract that has undergone classical chemical characterization, molecular docking, anti-diabetic, anticancer, antibacterial and antifungal studies used as traditional and complementary medicine. The study aimed to evaluate the Gas Chromatography–Mass Spectrometry (GC-MS), High-Performance Liquid Chromatography (HPLC), Liquid Chromatography–High Resolution Mass Spectrometry (LC-HRMS), anticancer, antidiabetic and spectral studies that are used to disclose phytochemicals and their medicinal properties. Methods: Infrared (IR), Ultraviolet (UV), 1H-NMR spectroscopic methods, GC-MS, HPLC and LC-HRMS instrumental detections were employed. Molecular docking, anti-diabetic, anticancer, antibacterial and antifungal studies were carried out using AutoDock 4.2.6. software program, α- Amylase method, Mosmann and Disk diffusion method respectively. Results: GC-MS and HPLC chromatogram of methanol extracts exhibited 12 and 6 peaks, respectively, confirming the presence of 12 phytoconstituents in Indus Viva I Pulse health drink (1I–12I). The total phenol and flavonoid content found in the extracted sample of Indus Viva I Pulse health drink were 0.16 and 0.36 mg/mL, respectively. It is a polyherbal formulation of brown-colored liquid of the UV spectrum in methanol is characteristic of an aromatic compound with λ max of 319 and 412 nm. IR spectrum gave peaks at 3,457, 2,922, 2,857, 2,121, 1,641, 1,055, 1,033 and 621 cm-1 indicating the presence of an alcoholic, alkyl, aldehyde or ketone, alkyne, carbonyl, anhydride and methyl group, respectively. Liquid Chromatography- High Resolution Mass Spectrometry (LC-HRMS) study gave these molecular weights (m/z): 1134. 7017, 1150.6475 and 1168.6797. Aromatic characteristics were confirmed through UV, IR, 1H-NMR, 13C-NMR and LC-HRMS spectral studies. 1H-NMR study indicates the presence of aromatic protons and methyl protons. Aqueous extract of Indus Viva I Pulse health drink was tested for Cytotoxicity Assay of human bone cancer (MG-63) cell lines and ptyalin prevent characters that exhibited a valid inhibitory value of 90.87% in 10 μg/mL for MG-63 and value of 74.41% in 500 μg/mL for alpha-amylase which is comparatively more efficient than the available standard drug. Cytotoxicity assay for human bone cancer (MG-63) cell lines and alpha-amylase both showed the inhibitory activity of IC50 68.95 and IC50 36.18 μg/mL, respectively. Conclusion: Based on the result of this research, it can be proposed that Indus Viva I Pulse health drink may serve as a potential remedy for human bone cancer (MG-63) cell line and that alpha-amylase inhibitory activity is proportional to the dose.

Production of Polyhydroxyalkanoates (PHA) by Probiotic Bacteria Bacillus tequilensis for Potentially Used as Drug Carrier

Introduction: Polyhydroxyalkanoate (PHA) is a biopolymer that can be produced by microorganisms from numerous low-cost carbon sources, making it an environmentally friendly material. This study was designed to utilize different food waste (household food waste, spent oils and spent coffee grounds) as nutrient source for the cultivation of microbes to produce polyhydroxyalkanoate (PHA). Methods: The bacterial strain Bacillus tequilensis was grown in 250 mL Erlenmeyer flask each containing 50 mL of mineral salt medium, 25 ml of nutrient broth inoculum and 20 g/L of household food waste, spent ground coffee and spent oils, respectively. The initial pH of the media was 7.0 and the cultured bacteria was incubated at 30 °C, 180 rpm for 72 h as a batch culture. The sample was then extracted and weight, and further analyzed using Fourier Transform Infrared Spectroscopy (FTIR) and High Performance Liquid Chromatography (HPLC). Results: B. tequilensis yielded PHA of 7 % to 8 % (g PHA/g dry cell weight) on average using medium containing household food waste, spent ground coffee and spent oils. FTIR analysis showed the peaks range between 1750-1730 cm-1 which belong to PHA functional groups such as C=O. HPLC chromatogram revealed that the retention time obtained from digested PHA was approximately 4.5 min which was similar to the standard of PHA. Conclusion: This enables the utilization of low-cost waste by probiotic B. tequilensis as a carbon source for the sustainable production of biodegradable PHA for a wide range of applications in medicine.

Antimalarial Potential of Fraction 5 from Ethanolic Leaves Extract of Artocarpus Altilis

Background: Artocarpus altilis leaf extract (AAL.E) was separated by VLC, and six fractions were obtained. Fraction 5 (AAL.E.5) showed antimalarial activity with an IC50 value of 3.71 µg/mL. Objective: This study aimed to determine the antimalarial activity of AAL.E.5 subfractions against P. falciparum, the mechanism of action against Plasmodium Falciparum Malate quinone oxidoreductase (PfMQO), and the active substances. Methods: The AAL.E.5 was separated by open-column chromatography and eluted with chloroform-methanol gradient elution in order of increasing polarity. The antimalarial activity of all subfractions was assessed using a lactate dehydrogenase (LDH) assay against P. falciparum and the mechanism of action of the PfMQO enzyme. The profiles of the most active subfractions were analyzed using High-Performance Liquid Chromatography (HPLC). Results: The separation of fraction 5 (AAL.E.5) yielded 11 subfractions (AAL.E.5.1–AAL.E.5.11). Screening antimalarial activity at 10 μg/mL in this subfraction showed that only five subfractions (AAL.E.5.6-AAL. E.5.10) inhibited P. falciparum and two subfractions (AAL.E.5.6 and AAL.E.5.10) inhibited the PfMQO enzyme. Only subfraction 6 (AAL.E.5.6) inhibited both, with IC50 values of 6.609 µg/mL and 20.34 µg/mL. The thin layer chromatography profile of AAL.E.5.6 revealed reddish-orange spots, indicating the presence of flavonoid compounds, and was also presumed from the UV-visible to HPLC chromatogram for band I in the 300 – 400 nm range and band II in the 240–285 nm range. Conclusion: Subfraction 6 has antimalarial activity against P. falciparum and is thought to have a mechanism of action in PfMQO. Based on the TLC, HPLC, and UV-Vis spectra, subfraction 6 was assumed to be a flavonoid.

Plant Mitochondrial-Targeted Gene Delivery by Peptide/DNA Micelles Quantitatively Surface-Modified with Mitochondrial Targeting and Membrane-Penetrating Peptides.

Plant mitochondria play essential roles in metabolism and respiration. Recently, there has been growing interest in mitochondrial transformation for developing crops with commercially valuable traits, such as resistance to environmental stress and shorter fallow periods. Mitochondrial targeting and cell membrane penetration functions are crucial for improving the gene delivery efficiency of mitochondrial transformation. Here, we developed a peptide-based carrier, referred to as Cytcox/KAibA-Mic, that contains multifunctional peptides for efficient transfection into plant mitochondria. We quantified the mitochondrial targeting and cell membrane-penetrating peptide modification rates to control their functions. The modification rates were easily determined from high-performance liquid chromatography chromatograms. Additionally, the gene carrier size remained constant even when the mitochondrial targeting peptide modification rate was altered. Using this gene carrier, we can quantitatively investigate the relationships between various peptide modifications and transfection efficiency and optimize the gene carrier conditions for mitochondrial transfection.

Influence of Voxelotor-hemoglobin complexes in the estimation of hemoglobin S levels by the current standard of care laboratory evaluation techniques.

Sickle cell disease is an inherited disorder characterized by the presence of sickle hemoglobin (HbS). The process of Hb molecule polymerization is a pivotal step in the sickling process. Voxelotor, a recently approved novel therapeutic agent, is known to interfere with polymerization. We aim to study the impact of Voxelotor on Hb variants analysis using high performance liquid chromatography (HPLC). We are reporting the impact of Voxelotor on Hb variants analysis using HPLC after an informed consent and medical research committee approval. Data was collected from eight patients who are enrolled in the GBT440-034OL study using electronic medical records, to evaluate the Hb levels, hemolytic markers and the clinical response. Our patients were well-balanced for gender, with a mean age of 31.1 years (19-50). Six patients showed a significant improvement in the Hb level, with reduced reticulocytes, bilirubin, LDH and an improved clinical outcome. Interestingly, these patients showed the appearance of a split band of Hb S and D on HPLC impacting significantly on HbS level. Two patients did not show any improvement on laboratory parameters, and no changes on their HPLC analysis. We report here eight patients on Voxelotor therapy, six of which showed improved hemolytic markers and anemia and demonstrated the appearance of HbD peak on the HPLC chromatogram. Therefore, the absence of HbD on HPLC or other laboratory methods for estimating HbS in patients on Voxelotor therapy, gives the clinician a possible hint regarding the patient's compliance with the drug.

Analytical assessment of modulated electric flux triggered degradation of chlorfenapyr and deltamethrin pesticides in guava fruits

The purpose of this study was to explore the new effective method and investigate the dissipation of chlorfenapyr and deltamethrin (DM) pesticides used in the treatment of guava fruit from tropical and sub-tropical areas of Pakistan. Five different solutions of varying concentrations of pesticides were prepared. This study involved the in-vitro and in-vivo analysis of modulated electric flux-triggered degradation as an efficient method for the safer degradation of selected pesticides. The Taser gun was used as a tool for providing different numbers of electrical shocks of million voltages to the pesticides present in guava fruit at different temperatures. The degraded pesticides were extracted and analyzed by High-performance liquid chromatography (HPLC). The HPLC chromatograms verified that significant dissipation of pesticides took place when these were exposed to 9 shocks at 37 °C, which proved the efficiency of this degradation method. More than 50% of the total spray of both pesticides was dissipated. Thus, modulated electrical flux-triggered degradation is one of the effective methods for pesticide degradation.

Extraction, physicochemical properties, and antioxidant activity of natural melanin from Auricularia heimuer fermentation.

Natural melanin from Auricularia heimuer have numerous beneficial biological properties, which were used as a safe and healthy colorant in several industries. In this study, single-factor experiments, Box-Behnken design (BBD), and response surface methodology (RSM) were employed to investigate the effects of alkali-soluble pH, acid precipitation pH, and microwave time on the extraction yield of Auricularia heimuer melanin (AHM) from fermentation. Ultraviolet-visible spectrum (UV-Vis), Fourier transform infrared (FT-IR) spectroscopy, scanning electron microscope (SEM), and high-performance liquid chromatography (HPLC) were used to analyze the extracted AHM. The solubility, stability, and antioxidant activities of AHM were also measured. The results showed that alkali-soluble pH, acid precipitation pH, and microwave time significantly affected the AHM yield, with the following optimized microwave-assisted extraction conditions: alkali-soluble pH of 12.3, acid precipitation pH of 3.1, and microwave time of 53 min, resulting in an AHM extraction yield of 0.4042%. AHM exhibited a strong absorption at 210 nm, similar to melanin from other sources. FT-IR spectroscopy also revealed that AHM exhibited the three characteristic absorption peaks of natural melanin. The HPLC chromatogram profile of AHM showed a single symmetrical elution peak with a 2.435 min retention time. AHM was highly soluble in alkali solution, insoluble in distilled water and organic solvents, and demonstrated strong DPPH, OH, and ABTS free radical scavenging activities. This study provides technical support to optimize AHM extraction for use in the medical and food industries.

Green and free hazardous substances of neem oil lotion in promising market sustainability

The demand for cosmetics has risen in recent years, resulting in mass production that does not adhere to any acceptable standards. Many of them contain potentially harmful substances, including hydroquinone, talcum, parabens, steroids, phthalates, fragrances, and heavy metals like lead and arsenic. The current study focused on producing neem oil lotion with free steroids, hydroquinone, and parabens. Prior to the testing, neem oil lotion was prepared by adding diluted neem oil, aloe vera and Vitamin E to the basic cream lotion. Thin Layer Chromatography (TLC) was used to analyze the steroid content. The spots that appeared were observed at different wavelengths; White Lamp (T-White), UV-254 nm, and UV-366 nm. The hydroquinone was run by High-Performance Liquid Chromatography (HPLC) and was observed under UV light at 254 nm. The parabens test was also done by HPLC under UV light at 280 nm. The analysis showed that the four types of steroids; dexamethasone, bethamethasone, triamcinolone acetonide, and hydrocortisone acetate, were not detected. Furthermore, no hydroquinone was observed, as proved by the HPLC chromatogram peak. In the parabens analysis, only methylparaben was detected in the sample with a concentration of 0.011 mg/mL, below the detection limit. The other three types of parabens; ethylparaben, propylparaben, and butylparaben, were not detected in the neem oil lotion formulation. The result showed that the prepared neem oil lotion was free from these hazardous substances; steroids, hydroquinone and parabens. It confirmed that the green source of neem leaf extract lotion had no hazardous substance and was safe to use.

Poloxamer and Chitosan-Based <i>In Situ</i> Gels Loaded with <i>Orthosiphon stamineus</i> Benth. Extracts Containing Rosmarinic Acid for the Treatment of Ocular Infections

Orthosiphon stamineus Benth. (OS) is a commonly used medicinal plant for curbing bacterial infections globally. This work aimed to fabricate poloxamer and chitosan-based in situ gels loaded with standardized aqueous-ethanolic OS leaf extracts and investigate their antimicrobial efficacy as a potential remedy against ocular infections. In situ gels containing 0.5% w/v OS extract prepared using cold dispersion method were subjected to physicochemical characterization, including in vitro-release studies. Antimicrobial efficacy was tested against Staphylococcus aureus, Micrococcus luteus, Escherichia coli, and Pseudomonas aeruginosa using agar diffusion method. Thin layer chromatography and high performance liquid chromatography chromatograms confirmed the presence of rosmarinic acid (RA) and sinensitin in OS extracts with same retention factor (0.26 and 0.49) and retention times (12.2 and 20.7 min) against reference standards. A homogenous brown coloured in situ gel exhibited low viscosity as a solution and increased viscosity in gel form at ocular temperature. The optimized formulations, P7 (21% P407/4% P188), P8 (21% P407/5% P188) and F5 (1.5% chitosan and 45% β-glycerophosphate) exhibited ideal ocular pH (7.27-7.46), phase transition at ocular temperature (33-37°C) and prolonged RA release up to 12 h. Formulation F5 showed an inhibition zone of 4.3 mm against M. luteus. Among all, formulation F5 alone exhibited modest antimicrobial activity against M. luteus. OS extracts at 5% and 10% were most active against tested bacteria however, loading them into in situ gels resulted in sedimentation. Hence, isolation of RA from OS extract is suggested before loading into formulations for a better antimicrobial activity.

Determination of HPLC Chromatogram Profile of Katuk (Breynia androgyna (L.) Chakrab. & N.P.Balakr) Plants from Ristoja's Results using Chemometric Analysis

The katuk plant was known as Sauropus androgynous (L.) Merr, but the name is changed to Breynia androgyna (L.) Chakrab. & N.P. Balakr since the publication of Chakrab's & N.P. Balakr in the 2012 Journal of Plant Taxonomists. The content of chemical compounds and secondary metabolites of katuk leaf are strongly influenced by different habitat or location. Therefore, the Center for Research and Development of Medicinal Plants and Traditional Medicines, National Institute of Health Research and Development (NIHRD) has conducted Research on medicinal plants and herbs (RISTOJA) results on 58 samples of katuk plants originating from 7 provinces and used by 13 ethnic groups in Indonesia. This study aims to obtain plant quality based on the description of the High-Performance Liquid Chromatography (HPLC) chromatogram profile, in order to obtain the suitability of the efficacy of medicinal plants with their benefits for the community in an area. Test using HPLC with a gradient mobile phase, mixture of acetonitrile and methanol for 60 minutes. The results of the chromatogram were analyzed chemometrically by using Principal Component Analysis (PCA) data interpretation. PCA results showed that from HPLC chromatograms at 254 nm and 366 nm, each gave 3 different clusters, namely Clusters A, B and C where each cluster has the same chromatogram profile of katuk plants. Cluster A which was identified at a wavelength of 254 nm was the most used cluster by 7 ethnic groups with a total of 28 Katuk samples. Similarly, the chromatogram at a wavelength of 366 nm with the largest cluster is cluster A which is used by 11 ethnic groups with a total of 45 samples of katuk plants. where each cluster had the same chromatogram profile of katuk Plants.

Quality Evaluation of Market Acacia catechu by Fingerprint-Chemical Pattern Recognition

Acacia catechu (L.f.) Willd, a leguminous plant, is included in the 2020 edition of the Chinese Pharmacopoeia and is mainly used to treat eczema, mouth ulcers, diarrhea, bruising, and traumatic hemorrhage. However, there are imported and domestic Acacia catechu samples available in China, and their quality and price are very different, which seriously affects the safety and stability of their clinical application. Importantly, there is no simple and effective method for identifying or classifying grades of Acacia catechu. In this study, 47 batches of commercial Acacia catechu were used for identifying or classifying grades of Acacia catechu using high performance liquid chromatography (HPLC) combined with chemometric analysis. Firstly, gradient elution was adopted with 0.05% phosphoric acid water (A)-methanol (B) as the mobile phase to establish chromatographic conditions. The HPLC chromatograms of 47 batches of Acacia catechu samples were analyzed by the “Similarity Evaluation System for Chromatographic Fingerprint of TCM” software (version 2012A). The common peaks of Acacia catechu were identified to evaluate the similarity. Based on the determination results of fingerprint chromatographic peak area, the quality of the collected Acacia catechu was evaluated by chemometric methods such as CA, PCA, and OPLS-DA. The results showed that the collected Acacia catechu samples were significantly divided into three categories. The first-class samples were all imported Acacia catechu except S9 sample, which was domestic Acacia catechu; the second-class samples were partly domestic Acacia catechu and partly imported Acacia catechu; and the third-class samples were all domestic Acacia catechu. Moreover, OPLS-DA of 47 batches of samples showed that the contents of catechin and the total contents of catechin and epicatechin could be used as key indicators for assessing the quality of Acacia catechu. The developed HPLC fingerprint and quantitative analysis method of multi-indicator components can be used for classification and quality evaluation of market Acacia catechu, which has a significant reference value for developing Acacia catechu grade quality standards.

Ultrasound Assisted Extraction and Characterization of Bioactives From Verbascum thapsus Roots to Evaluate Their Antioxidant and Medicinal Potential.

Worldwide, Verbascum thapsus is used for the treatment of various ailments owing to the presence of bioactives of therapeutic potential. Current study was planned to extract bioactives from V thapsus roots using methanol and water as solvents under stimulated effect of ultrasonic waves and characterize them to evaluate their potential benefits. Proximate analysis explored the presence of significant contents of protein, fats, fiber, organic and inorganic minerals. Fourier transform infrared spectra and high-performance liquid chromatography chromatogram indicated the presence of different phytochemicals having antioxidant potential as evidenced by total phenolic contents, total flavonoids content and 2,2-diphenyl-1-picrylhydrazyl activity of both extracts. Both extracts displayed excellent antimicrobial potential against Staphylococcus aureus (S aureus) and Fusarium Solani (F solani). Aqueous and methanolic extracts exhibited higher inhibition of biofilm made by Bacillus subtilis (B subtilis) as 55.09 and 61.58%, respectively in comparison to biofilm of Escherichia coli (E coli) as 48.11 and 36.51%, respectively. Methanol extract exhibited anti-amylase activity (IC50 5.26 ± .31 μg/mL) with an inhibition rate of 68.11% whereas IC50 of aqueous extract was 6.59 ± .53 μg/mL with an inhibition rate of 63.53%. Inhibitory potential against α-glucosidase (IC50 3.70 ± .94 ppm) was demonstrated by methanol extract in comparison to aqueous extract (IC50 7.58 ± .15). The study concluded that V thapsus roots have significant medicinal potential due to the presence of variety of bioactive molecules and can be used in pharmaceutical preparations.

The components transitive regularity of three dosage forms of Liuwei Dihuang Fufang

ObjectiveTo explore the transitive regularity of holistic constituents from the crude slices of the medicinal raw materials (MCS) to the formula granules (FG), fufang decoction (FD), and finally, the concentrated pills (CP) of Liuwei Dihuang Fufang (六味地黄复方, LWDHF). MethodsSamples for MCS, FG, FD, and CP of LWDHF were obtained, and a fingerprint database was established using high-performance liquid chromatography (HPLC), by separating the samples in an XB-C18 column and analyzing the transitive regularity of components using the total quantum statistical moment (TQSM), including total quantum zero moment (AUCT), total quantum first moment (MRTT), total quantum second moment (VRTT), and its similarity approach. The AUCT, MRTT, and VRTT were calculated based on the representative HPLC chromatograms of FG, FD, and CP of LWDHF. ResultsAUCT of FG, FD, and CP of LWDHF was 71 804, 46 553, and 144 646 μV s, respectively; MRTT was 14.43, 14.54, and 18.85 min, respectively; and VRTT was 106.98, 112.84, and 269.12 min2, respectively. Comparing the similarity of FG/FD, FG/CP and FD/CP of LWDHF, the TQSM similarity values were 98.66%, 76.62%, and 75.37%, respectively, whereas the traditional similarity evaluation values were 98.68%, 85.43%, and 85.60%, respectively. ConclusionThe results perform little distinction in the total composition between FG and FD, whereas some distinction existed between FD and CP. Experimental evidence, therefore indicates that FG could be used as the alternative of MCS in clinical applications.

Degradation of Imidacloprid Residue in Cabbage (Brassica oleracea) Using Advanced Oxidation Processes and Its Analyzing by Spectrophotometer and HPLC

Imidacloprid is a synthetic pesticide used to kill and control insects by cabbage farmers. Consuming cabbage contaminated with imidacloprid can cause health problems such as chronic kidney, reproductive disorders and cancer. The parameters for degradation of imidacloprid residue in cabbage were processing time (5, 10, 15, 20, and 25 min), water volume (50, 100, 150, and 200 mL), and sample mass (25, 50, 75, and 100 g). Degradation of imidacloprid residue in cabbage using Advanced Oxidation Processes (AOPs) methods such as ozonolysis, sonolysis, and sonozolysis were compared. Methods of ozonolysis, sonolysis, and sonozolysis have performed using an ozonicator (ozone dose: 400 mg h-1), sonicator (35 kHz), and a combination of both. Using spectrophotometers UV-Vis and High-Performance Liquid Chromatography (HPLC) measurements showed a decrease in imidacloprid concentration after degradation. The degradation percentage of the imidacloprid residue with ozonolysis was higher than that of sonolysis and sonozolysis. The percentage of imidacloprid degradation by ozonolysis in 50 g cabbage at 100 mL of water increased to 82.53% for 15 min. The HPLC chromatogram showed no new peaks that formed after degradation. The imidacloprid residue’s degradation kinetics were then investigated. For all AOPs methods, the kinetic analysis demonstrated that imidacloprid degradation fit within a first-order kinetic model. The kinetic data showed that ozonolysis degraded imidacloprid with a half-life (t1/2) of 15.22 min, shorter than sonolysis and sonozolysis.

HPLC analysis of carotenoids from bacteria.

Bacteria synthesize different carotenoid structures. A powerful method for the determination of carotenoid profiles of bacteria, including quantification, is high performance liquid chromatography (HPLC). This chapter explains the basics of this method and its application, and provides instruction on how to proceed with HPLC. A basic HPLC system for bacterial carotenoids is presented which allows tentative identification and quantification. The individual analysis steps including extraction, sample preparation and HPLC separation are described in detail. HPLC examples of structurally diverse carotenoids from different bacterial phyla are displayed demonstrating the applicability of this method. In addition, information on spectroscopic properties useful for identification is included. Finally, a procedure for carotenoid quantification from the HPLC chromatogram is described.

Antioxidant and Anti-Inflammatory Activities of Ethanol Extract and its Fractions from Acer tegmentosum Stem

In this study, the first purpose was to determine total phenolic, total flavonoid and total terpenoid content of the Acer tegmentosum ethanol extract and four different types of fractions, including chloroform, ethyl acetate, n-butanol and water. Among the samples, Acer tegmentosum ethanol extract and fraction of ethyl acetate showed the highest total phenolic, total flavonoid and total terpenoid content. The second purpose was to evaluate the antioxidant activity of the samples assessed by in vitro methods such as scavenging capacity of 2,2-diphenyl-1-picryl-hydrazyl-hydrate and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid free radicals. Especially, Acer tegmentosum ethanol extract and fraction of ethyl acetate showed the strongest scavenging capacity with the lowest half maximal scavenging capacity values of these free radicals. The third purpose was to evaluate the anti-inflammatory activity in the lipopolysaccharide-stimulated Ralph and William’s cell line 264.7 macrophages treated with various concentrations of Acer tegmentosum ethanol extract and fraction of ethyl acetate, and no cytotoxic effect on the macrophages was observed at the concentration of 100 μg/ml of this extract. It suggested that this extract suppresses the activation of macrophages to secrete both pro-inflammatory cytokines (interleukin-1 beta, tumour necrosis factor alpha, inducible nitric oxide synthase and interleukin-6) and enzyme (inducible nitric oxide synthase) and anti-inflammatory cytokine (interleukin-10). The fourth purpose was to identify some compounds in a sample responsible for the antioxidant and anti-inflammation activities. In high-performance liquid chromatography chromatograms, the peak retention times detected from Acer tegmentosum ethanol extract and fraction of ethyl acetate in comparison with those from reference standards identified six compounds such as gallic acid, salidroside, (-)-epigallocatechin, (+)-catechin, scopoletin and trans-ferulic acid. It suggested that these compounds are responsible for antioxidant and anti-inflammation activities. Taken together, Acer tegmentosum seems to be one of the most promising species as a therapeutic source against diseases.

α-glucosidase inhibitors from Syzygium polyanthum (Wight) Walp leaves as revealed by metabolomics and in silico approaches

Ethnopharmacological relevanceSyzygium polyanthum (Wight) Walp leaves are traditionally used to cure diabetes in many regions of Indonesia. Traditional use involves boiling the leaves until the water is reduced to half volume, and then the decoction is taken 1–2 times daily. Despite several studies reporting the antidiabetic activity of this plant, bioactive compounds have not been well identified. Aim of the studyIndonesia is one of the countries with the highest diabetes cases, particularly type 2 diabetes mellitus (T2DM). Few people have access to modern medicinal treatment; thus, the role of antidiabetic traditional medicine has become increasingly important. This research aimed to identify α-glucosidase inhibitors from S. polyathum leaves using a metabolomics approach. When the active compounds of S. polyathum are properly identified, the quality of the herb can be more easily controlled. Materials and methodsThe dried leaves of S. polyanthum were extracted by a comprehensive extraction method using a solvent combination of n-hexane, acetone, and water in a gradient, resulting in a total of 42 fractions. All fractions were subjected to an in vitro α-glucosidase inhibition test and chemical profile analysis using Nuclear Magnetic Resonance (NMR) and high performance liquid chromatography (HPLC). Orthogonal projection least square (OPLS) analysis was used to correlate the two data to identify NMR signals, and HPLC chromatogram peaks correlated to the activity. 2D NMR and ultra-high-performance liquid chromatography coupled to high-resolution mass spectrometry (UHPLC-HRMS) analyses were also used to give more precise compound identification. The activity of the identified active compounds was confirmed by an in silico technique. Results and discussionThe results of the α-glucosidase activity test showed that the most active fractions were obtained from solvents with medium polarity: Fractions 9 and 10 (F9 and F10), obtained from gradient acetone-water 4:1 and 3:2, respectively. The IC50 values of F9 and F10 were 24.8 and 31.8 μg/mL, respectively. NMR data showed that F9 had more intense and diverse signals in the aromatic region than F10. OPLS analysis results showed that some typical flavonoid signals abundant in F9 positively correlated with α-glucosidase activity. 2D NMR and UHPLC-HRMS analysis of F9 led to the conclusion that these signals could be attributed to myricetin-3-O-rhamnoside (myricitrin) and epigallocatechin-3-gallate (EGCG). In silico analysis confirmed these results, as myricitrin and EGCG had binding energies resembling acarbose as a positive control (-8.47, -8.19, and -10.13, respectively). ConclusionsNMR and HPLC-metabolomics successfully identified myricitrin and EGCG as α-glucosidase inhibitors from S. polyanthum leaves, and docking analysis validated their inhibitory activity. The results of this study justified the traditional use of S. polyanthum as an antidiabetes herbal.