Exploring the Anti-inflammatory Effects of Protopine Total Alkaloids of Macleaya Cordata (Willd.) R. Br.

Macleaya cordata (Willd). R. Br. is a Chinese medicinal plant commonly used externally to treat inflammatory-related diseases such as arthritis, sores, and carbuncles. This study aimed to evaluate the anti-inflammatory activity of protopine total alkaloids (MPTAs) in Macleaya cordata (Willd.) R. Br. in vivo tests in rats with acute inflammation showed that MPTA (2.54 and 5.08 mg/kg) showed significant anti-inflammatory activity 6 h after carrageenan injection. Similarly, MPTA (3.67 and 7.33 mg/kg) showed significant anti-inflammatory activity in the mouse ear swelling test. In addition, the potential mechanisms of the anti-inflammatory effects of MPTA were explored based on network pharmacology and molecular docking. The two main active components of MPTA, protopine and allocryptopine, were identified, and the potential targets and signaling pathways of MPTA's anti-inflammatory effects were initially revealed using tools and databases (such as SwissTargetPrediction, GeneCards, and STRING) combined with molecular docking results. This study provides the basis for the application of MPTA as an anti-inflammatory agent.

Safety assessment of MPTA: An oral acute and 90-day sub-chronic toxicity study in Sprague-Dawley rats.

MPTA is a novel extract product derived from Macleaya cordata (Willd.) R. Br., which has good anti-inflammatory and antioxidant activity. The aim of this study was to investigate the acute oral toxicity and 90-day sub-chronic oral toxicity of MPTA. In the acute toxicity study, 50 SD rats of both sexes were randomly divided into 5 groups and dosed in a gradient from 197.53 mg/kg to 1000.00 mg/kg bw. Toxic effects were observed up to 14 days and LD50 was calculated. In a subchronic toxicity test, male and female SD rats were orally dosed repeatedly with 96.40, 19.28, 3.86 mg/kg bw of MPTA for 90 days. In addition, a control group was set up in the subchronic study. The acute toxicity test showed that the oral LD50 of MPTA was 481.99 mg/kg with a 95% confidence interval of 404.24-574.70 mg/kg. MPTA did not appear to induce toxic effects in the longer term in terms of food and water consumption, weight gain, haematological and clinical biochemical parameters and pathological examination. The first data on the potential toxicity of MPTA was provided to highlight the safety of short-term to longer-term oral administration of MPTA, and the experimental results yield and establish a NOEAL of 96.40 mg/kg/d for MPTA.

Pharmacokinetics of sanguinarine, chelerythrine, and their metabolites in broiler chickens following oral and intravenous administration.

Sanguinarine (SA) and chelerythrine (CHE) are the main active components of the phytogenic livestock feed additive, Sangrovit®. However, little information is available on the pharmacokinetics of Sangrovit® in poultry. The goal of this work was to study the pharmacokinetics of SA, CHE, and their metabolites, dihydrosanguinarine (DHSA) and dihydrochelerythrine (DHCHE), in 10 healthy female broiler chickens following oral (p.o.) administration of Sangrovit® and intravenous (i.v.) administration of a mixture of SA and CHE. The plasma samples were processed using two different simple protein precipitation methods because the parent drugs and metabolites are stable under different pH conditions. The absorption and metabolism of SA following p.o. administration were fast, with half-life (t1/2 ) values of 1.05 ± 0.18 hr and 0.83 ± 0.10 hr for SA and DHSA, respectively. The maximum concentration (Cmax ) of DHSA (2.49 ± 1.4 µg/L) was higher that of SA (1.89 ± 0.8 µg/L). The area under the concentration vs. time curve (AUC) values for SA and DHSA were 9.92 ± 5.4 and 6.08 ± 3.49 ng/ml hr, respectively. Following i.v. administration, the clearance (CL) of SA was 6.79 ± 0.63 (L·h-1 ·kg-1 ) with a t1/2 of 0.34 ± 0.13 hr. The AUC values for DHSA and DHCHE were 7.48 ± 1.05 and 0.52 ± 0.09 (ng/ml hr), respectively. These data suggested that Sangrovit® had low absorption and bioavailability in broiler chickens. The work reported here provides useful information on the pharmacokinetic behavior of Sangrovit® after p.o. and i.v. administration in broiler chickens, which is important for the evaluation of its use in poultry.

Medicinal plants of the genus Macleaya (Macleaya cordata, Macleaya microcarpa): A review of their phytochemistry, pharmacology, and toxicology.

In the genus Macleaya, Macleaya cordata and Macleaya microcarpa have been recognized as traditional herbs that are primarily distributed in China, North America, and Europe and have a long history of medicinal usage. These herbs have been long valued and studied for detumescence, detoxification, and insecticidal effect. This review aims to provide comprehensive information on botanical, phytochemical, pharmacological, and toxicological studies on plants in the genus Macleaya. Plants from the genus of Macleaya provide a source of bioactive compounds, primarily alkaloids, with remarkable diversity and complex architectures, thereby having attracted attention from researchers. To date, 291 constituents have been identified and/or isolated from this group. These purified compounds and/or crude extract possess antitumor, anti-inflammatory, insecticidal, and antibacterial activities in addition to certain potential toxicities. Macleaya species hold potential for medicinal applications. However, despite the pharmacological studies on these plants, the mechanisms underlying the biological activities of active ingredients derived from Macleaya have not been thoroughly elucidated to date. Additionally, there is a need for research focusing on in vivo medical effects of Macleaya compounds and, eventually, for clinical trials.

[Research strategies for feed additives and veterinary medicines from side products of Chinese medicine resources industrialization].

The global antimicrobial resistance has been a big challenge to the human health for years. It has to make balance between the safety of animal products and the use of antimicrobials in animal husbandry. Any methods that can minimize or even phase out the use of antimicrobials in animal husbandry should be encouraged. We herein describe the research strategies for feed additives and veterinary medicines from the side products of Chinese medicine resources industrialization. Killing two birds with one stone-besides the major purposes, the rational utilization of non-medicinal parts and wastes of industrialization of Chinese herbal medicines is also achieved under the proposed strategies.

[A new 6,6a-offbenzophenanthridine alkaloid from Macleaya cordata].

A new alkaloid was isolated from the leaves of Macleaya cordata with 95% ethanol extracted and its isolation was by column chromatography and preparation HPLC. The new structure was elucidated as 6'-hydroxy-2',3'-dimethoxyarnottianamide on the basis of its spectroscopic date.

Mass spectrometry-guided isolation of two new benzoquinoline alkaloids from Macleaya cordata.

Two new alkaloids, named 2,3-methylenedioxy-7,10-dimethyl-7,8,9,10-tetrahydro-benzoquinoline (1) and 2,3-methylenedioxy-7,10-dimethyl-8-carboxyl-benzoquinoline (2), were detected primarily from the fruits of Macleaya cordata by their different fragmentation pathways. And then isolation of the two compounds was performed by column chromatography and preparative HPLC under the guiding of mass spectrometry. Finally, their structures were determined by spectroscopic analysis.

Structural speculation and identification of alkaloids in Macleaya cordata fruits by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry combined with a screening procedure.

RATIONALE: Alkaloids with significant therapeutic effects are the main active constituents of Macleaya cordata, which is a perennial herb plant in the Papaveraceae family. A systematic and novel method for speculating and identifying the structures of alkaloids in M. cordata fruits by high-performance liquid chromatography/quadrupole-time-of-flight mass spectrometry (HPLC/Q-TOF-MS) with a screening procedure was reported. METHODS: Investigation of mass spectral fragmentation of alkaloids was carried out based on the tandem mass spectrometry (MS/MS) data analyses of eight reference substances. The skeletons of alkaloids were determined by their ultraviolet spectra (UV) and MS/MS data. The substituent groups of the alkaloids were acquired through a screening procedure developed in our laboratory and MS/MS data. The substituent linkage sites were deduced by MS/MS fragmentation behavior, as well as biosynthetic pathways of related alkaloids. RESULTS: The structures of 21 alkaloids were speculated in this study, 10 of which were reported for the first time in M. cordata. Furthermore, benzyltetrahydroisoquinoline and N-methyltetrahydroprotoberberine-type alkaloids were discovered, which indirectly proved that the biosynthetic pathways of benzophenanthridine alkaloids reported in Eschscholtzia california existed in M. cordata as well. CONCLUSIONS: HPLC/Q-TOF-MS combined with a screening procedure was a systematic and reliable method for speculating and elucidating the structures of alkaloids. This study might be useful for the identification of other compounds in herbal medicines.

Optimization of microwave-assisted extraction of protopine and allocryptopine from stems of Macleaya cordata (Willd) R. Br. using response surface methodology.

The purpose of the research was to investigate the multiple response optimizations for the extraction of protopine and allocryptopine from the stems of Macleaya cordata (Willd) R. Br. by using microwave-assisted extraction (MAE). A three-level, three-factor Box-Behnken design of response surface methodology was used to develop response model, and desirability function was employed to optimize the effects of main extraction parameters. Three variables, ethanol concentration (20-80%, v/v), extraction temperature (30-70 degrees C) and solvent/solid ratio (10:1 to 30:1, mL/g), were investigated in this study. The results showed that the optimum parameters of MAE were ethanol concentration of 45.2 % (v/v), extraction temperature of 54.7 degrees C and solvent/solid ratio of 20.4:1 (mL/g). Under these conditions, the extraction yields of protopine and allocryptopine were 89.4 and 102.0%, respectively, and the extracta sicca yield was 12.5%. The combination use of response surface methodology, Box-Behnken design and the appropriate desirability function could provide an insight into a lab-scale MAE process, and help to develop procedures for commercial production of active ingredients from medical plants.

Analysis of alkaloids in Macleaya cordata (Willd.) R. Br. using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry.

A method for the analysis of alkaloids in Macleaya cordata (Willd.) R. Br. using high-performance liquid chromatography with diode array detection and electrospray ionization mass spectrometry (HPLC-DAD-ESI/MS) was developed. Using protopine (PRO), allocryptopine (ALL), sanguinarine (SA), and chelerythrine (CHE) as the model components, different columns for the separation and different mobile phases for the signal intensities of alkaloids in ESI/MS were investigated, respectively. The results showed that good separation and high signal intensities can be obtained on a high carbon loading (17%) reversed-phase C(18) column with 30 mM formic acid in mobile phase for the analysis of alkaloids. Under the optimal separation condition and UV detection (284 nm), linearity of the six alkaloids was obtained over concentration range from 0.05 to 100.00 microg/ml. The limit of detection (LOD) was 1.62, 1.87, 1.79, 1.76, 1.10, and 0.94 ng/ml for SA, CHE, PRO, ALL, dihydrosanguinarine (DHSA), and dihydrochelerythrine (DHCHE), respectively. The LODs with ESI/MS detection were lower three orders of magnitude than those obtained with UV detection. The proposed method could be used to control quality of the raw materials of the herb more comprehensively.

Investigation of phenolic constituents in Echinacea purpurea grown in China.

Echinacea is a North American native medicinal herb. In 1990 s, it was introduced in China. Nowadays, Echinacea is growing successfully in a number of places in China, and has been used as a crude drug. However, the phytochemical variation in the plant grown in China has not been studied. In this study, the contents of total phenolics and caffeic acid derivatives in aerial parts and roots of Echinacea purpurea grown in China were investigated by high-performance liquid chromatography (HPLC) and colorimetric analysis. The effects of different drying methods on the components were also studied. The results show that the content of caffeic acid derivatives in E. purpurea reached its highest in the middle stage of full blossoming. The content of caffeic acid derivatives in fresh raw material was generally higher than that in dried raw material. There was no significant difference in the content of caffeic acid derivatives among three geographical populations of E. purpurea. Furthermore, the developmental pattern of total phenolics in E. purpurea was the same as that of caffeic acid derivatives. The stage of mid-bloom is an optimal harvesting period for both caffeic acid derivatives and total phenolics. In addition, the results show that the fresh raw material is the optimal material for pharmaceutical purposes, and that the optimal pharmaceutical parts are the roots, leaves and flowers.

[Determination of benzopyrene in red clover extract by high performance liquid chromatography with spectrofluorescence detector].

OBJECTIVE: To establish a high performance liquid chromatography method with spectrofluorescence detector for the quantitative analysis of benzopyrene in red clover extract. METHOD: Analysis was performed on Hypersil C18 column (4. 6 mm x 200 mm, 5 microm) with acetonitrile and water (70: 30) as mobile phase; The excitation wavelength and emission wavelengh were 368 nm and 405 nm, respectively; Flow rate was 1.0 mL x min(-1). RESULT: The linear range was 0.93-37.2 ng x mL(-1), and r = 0.9992 (n = 5). The average recovery of benzopyrene was 85.2% (n = 9). CONCLUSION: The method was convenient and accurate with good recovery.

[Simultaneous determination of resveratrol, emodin, chrysophanol, physcion, in root of Polygonam cuspidatum and its extract by HPLC].

OBJECTIVE: To establish a HPLC method for simultaneous determination of resveratrol, emodin, chrysophanol, physcion in root of Polygonum cuspidatum and its extract. METHOD: The separation was performed in a Hypersil ODS C18 column with a mobile phase of Acetonitrile-1%HAc. The flow rate was 1.0 mL x min(-1) and detection wavelength was at 287 nm. RESULT: The mean recovery and RSD of resveratrol, emodin, chrysophanol, physcion was 101.8%, RSD 1.7%; 96.2%, RSD 1.4%; 99.7%, RSD 1.0%; 98.60%, RSD 0.97% respectively. CONCLUSION: The method is accurate, simple and reliable. It can be used for quality control of P. cuspidatum and its extract.

Simultaneous analysis of caffeic acid derivatives and alkamides in roots and extracts of Echinacea purpurea by high-performance liquid chromatography-photodiode array detection-electrospray mass spectrometry.

High-performance liquid chromatography (HPLC) coupled with UV photodiode-array detection and electrospray ionization mass spectrometry was developed for the simultaneous analysis of caffeic acid derivatives and alkamides in the roots and extracts of Echinacea purpurea. Caffeic acid derivatives and alkamides produced very abundant peaks in the total ion current chromatogram during negative and positive cone voltage switching. Cichoric acid and the isomer pair, dodeca-2E,4E,8Z,10E/Z-tetraenoic acid isobutylamide, were used as a standard for quantification of caffeic acid derivatives and alkamides in E. purpurea. This novel method surpasses previously published ones in product quality control and providing the HPLC chromatographic fingerprints of biological active components in E. purpurea.