Isolation of flavonoids from Tagetes patula (French Marigold) flowers: Cytotoxic and oxidant activity in human cervical carcinoma cells / Aislación de flavonoides de las flores de Tagetes patula (cempasúchil francés): Actividad citotóxica y oxidante en células de carcinoma cervical humano

T agetes patula , known as French Marigold, belongs to the family Asteraceae. Human papillomavirus infection is considered one of the causes of cervical cancer. This study assessed the cytotoxic activity and intracellular oxidative capacity of compounds isolated from extract of T. patula flowers as anti - cancer cervical agents. Fraction F6 of n - butanol extract was subjected to column chromatography and HPLC - ESI - MS. The isolated compo unds of T. patula were used to examine cytotoxic activity and the production of total reactive oxygen species in SiHa and HeLa cells; the cells were also characterized using scanning electron microscopy. Patulitrin was cytotoxic to SiHa and HeLa cells. An increase in ROS production was observed at different times of treatment of cells with patuletin and patulitrin. Scanning electron microscopy showed morphological changes in SiHa and HeLa cells. Thus, compounds isolated from T. patula have great treatment p otential against cervical cancer.

Tagetes patula , conocida como cempasúchil francés, pertenece a la familia Asteraceae. La infección por el virus del papiloma humano se considera una de las causas del cáncer cervical. En este estudio, se evaluó la actividad citotóxica y la capacidad oxidativa intracelular de los compuestos aislados del extracto de las flores de T. patula como agentes anticancerígenos cervicales. La fracción F6 del ext racto de n - butanol se sometió a cromatografía en columna y HPLC - ESI - MS. Los compuestos aislados de T. patula se utilizaron para examinar la actividad citotóxica y la producción total de especies reactivas de oxígeno en las células SiHa y HeLa; las células también se caracterizaron mediante microscopía electrónica de barrido. Patulitrina resultó citotóxica para las células SiHa y HeLa. Se observó un aumento en la producción de ROS en diferentes momentos del tratamiento de las células con patuletina y patulit rina. La microscopía electrónica de barrido mostró cambios morfológicos en las células SiHa y HeLa. Por lo tanto, los compuestos aislados de T. patula tienen un gran potencial de tratamiento contra el cáncer cervical.

Phytochemical Screening and in vitro Anticancer Activity of Some Medicinal Plants Grown in the Kingdom of Saudi Arabia / Cribado Fitoquímico y Actividad Anticancerígena in vitro de Algunas Plantas Medicinales Cultivadas en el Reino de Arabia Saudita

SUMMARY: This study evaluated the phytochemical screening, antioxidant capacity, and in vitro anticancer activities of four plants namely, Gypsophila capillaris, Anabasis lachnantha, Haloxylon salicornicum, and Horwoodia dicksoniae which belong to four different families: Caryophyllaceae, Amaranthaceae, Chenopodiaceae, Brassicaceae, respectively. The total phenolics, anthocyanins, saponins, total antioxidant capacity (TAC), and DPPH assays were determined by spectrophotometer. In vitro anticancer activity was assessed using two human cancer cell lines; hepatocellular carcinoma (HepG-2) and breast adenocarcinoma (MCF-7) to estimate the inhibition concentration 50 % (IC50). The results showed that H. dicksoniae has the highest concentrations of phenolics and saponins, while H. salicornicum has the highest DPPH. The highest concentration of TAC was found in G. capillaries. Among the tested extracts, G. capillaries and H. salicornicum have the potential activity against MCF-7 and HepG-2 cell lines in vitro. The content of polyphenols in G. capillaries was profiled by high-performance liquid chromatography (HPLC). The highest concentration among the phenolic compounds was chlorogenic (60.8 µg/ml) while the highest concentration among the flavonoid compounds was hesperidin (1444.92 µg/ml). In summary, G. capillaries and H. salicornicum extracts have potent anticancer activity against HepG-2 and MCF-7 cell lines.

Este estudio evaluó la detección fitoquímica, la capacidad antioxidante y las actividades anticancerígenas in vitro de cuatro plantas, Gypsophila capillaris, Anabasis lachnantha, Haloxylon salicornicum y Horwoodia dicksoniae, que pertenecen a cuatro familias diferentes: Caryophyllaceae, Amaranthaceae, Chenopodiaceae y Brassicaceae, respectivamente. Los ensayos de fenólicos totales, antocianinas, saponinas, capacidad antioxidante total (TAC) y DPPH se determinaron mediante espectrofotómetro. La actividad anticancerígena in vitro se evaluó utilizando dos líneas celulares de cáncer humano; carcinoma hepatocelular (HepG-2) y adenocarcinoma de mama (MCF- 7) para estimar la concentración de inhibición del 50 % (IC50). Los resultados indicaron que H. dicksoniae tiene las concentraciones más altas de fenólicos y saponinas, mientras que H. salicornicum tiene el DPPH más alto. La mayor concentración de TAC se encontró en G. capillaries. Entre los extractos probados, G. capillaries y H. salicornicum tienen actividad potencial contra líneas celulares MCF-7 y HepG-2 in vitro. El contenido de polifenoles en G. capillaries se perfiló mediante cromatografía líquida de alta resolución (HPLC). La concentración más alta entre los compuestos fenólicos fue clorogénica (60,8 µg/ml), mientras que la concentración más alta entre los compuestos flavonoides fue la hesperidina (1444,92 µg/ml). En resumen, los extractos de Gypsophila capillaris y H. salicornicum tienen una potente actividad anticancerígena contra las líneas celulares HepG-2 y MCF-7.

Validation of an analytical method by HPLC for the quantification of yangambin from the Ocotea duckei species / Validación de un método analítico por HPLC para la cuantificación de yangambin de la especie Ocotea duckei

Ocotea duckei , known as Louro - de - cheiro, belongs to the Lauraceae family and presents lignoid yangambine (YAN) as the main plant marker. This work aimed to develop and validate an analytical method by high performance liquid chromatography for the quantification of YAN. The sample used was the crude eth anolic extract (CEE) obtained from aerial parts. In the developed method, a C18 column was used. The mobile phase was composed of acetonitrile and water (45:55), whereas the method parameters included mobile phase flow rate at 0.8 mL/min, oven temperature at 40°C, and monitoring at 205 nm. In the validation, the parameters of selectivity, linearity, precision, accuracy, robustness, limits of detection and quantification were evaluated. As a result, the developed method is in accordance with the guidelines f or validation of analytical methods and presented satisfactory chromatographic parameters for YAN determination. Thus, the present analytical methodology can be applied in the quality control of O. duckei raw materials.

Ocotea duckei , conocida como Louro - de - cheiro, pertenece a la familia Lauraceae y presenta la yangambina lignoide (YAN) como principal marcador vegetal. Este trabajo tuvo como objetivo desarrollar y val idar un método analítico por cromatografía líquida de alta resolución para la cuantificación de YAN. La muestra utilizada fue el extracto etanólico crudo (EEC) obtenido de partes aéreas. En el método desarrollado se utilizó una columna C18. La fase móvil c onsistió en acetonitrilo y agua (45:55), mientras los parámetros del método incluyeron el caudal de la fase móvil a 0,8 m L /min, la temperatura del horno a 40°C y la monitorización a 205 nm. En la validación se evaluaron los parámetros de selectividad, line alidad, precisión, exactitud, robustez, límites de detección y cuantificación. Como resultado, el método desarrollado está de acuerdo con las pautas para la validación de métodos analíticos y presentó parámetros cromatográficos satisfactorios para la deter minación de YAN. Por lo tanto, la presente metodología analítica se puede aplicar en el control de calidad de las materias primas de O. duckei.

RP-HPLC method for rhein quantification in Cassia fistula L. (Fabaceae) leaves

Objectives@#The aim of this study is to establish a Reversed Phase – High Performance Liquid Chromatographic (RP-HPLC) method for the quantification of Rhein from Cassia fistula L. leaves.@*Methods@#A Shimadzu system equipped with a C18 Column (150 x 4.6 mm, 5 μm) with an isocratic elution of Acetonitrile (solvent A) and 0.1% trifluoroacetic acid aqueous solution (solvent B) (Merck, 1.08178.0050) with a 55:45 ratio, respectively and a flow rate of 1.0 mL/min and sample injection of 10 μL detection was done at 230 nm. Standard solution of Rhein (Chengdu Biopurify) was prepared for method development. This study was validated using the guidelines set under “ICH Topic Q2 R2 or the Validation of Analytical Procedures”. Procedures for linearity, precision, accuracy, limit of detection, and limit of quantitation were performed.@*Results@#The retention time of Rhein standard was determined at 5.10 minutes. LOD and LOQ were determined to be 1.278 mcg/mL and 3.872 mcg/mL, respectively with good linearity (R2 ≥0.996) with a linear range of 2.5-20 ug/mL of the Rhein standard. The accuracy of the method was determined based on % recovery method and ranged from 94.75%-100.32% (intraday, n=3) with %RSD of 0.71. The intraday precision %RSD was 2.92 (n=6) while interday precision %RSD was 3.75 (n=3). The method was able to check the Rhein quantity among 10 samples of Cassia fistula L. leaves from different locations in the Philippines.@*Conclusion@#The method was found to be sensitive and accurate for the quantification of Rhein. The method was found to be useful for the quantification of the amount of Rhein and can be used as a Quality Control tool for the assessment of Cassia fistula.

Poly-ε-caprolactone based nanoparticles for delivery of genistein in melanoma treatment

We developed poly-ε-caprolactone (PCL)-based nanoparticles containing D-α-tocopherol polyethylene glycol-1000 succinate (TPGS) or Poloxamer 407 as stabilizers to efficiently encapsulate genistein (GN). Two formulations, referred to as PNTPGS and PNPol, were prepared using nanoprecipitation. They were characterized by size and PDI distribution, zeta potential, nanoparticle tracking analysis (NTA), GN association (AE%), infrared spectroscopy (FT-IR), and differential scanning calorimetry (DSC). PNTPGS-GN exhibited a particle size of 141.2 nm, a PDI of 0.189, a zeta potential of -32.9 mV, and an AE% of 77.95%. PNPol-GN had a size of 146.3 nm, a better PDI than PNTPGS-GN (0.150), a less negative zeta potential (-21.0 mV), and an AE% of 68.73%. Thermal and spectrometric analyses indicated that no new compounds were formed, and there was no incompatibility detected in the formulations. Cellular studies revealed that Poloxamer 407 conferred less toxicity to PCL nanoparticles. However, the percentage of uptake decreased compared to the use of TPGS, which exhibited almost 80% cellular uptake. This study contributes to the investigation of stabilizers capable of conferring stability to PCL nanoparticles efficiently encapsulating GN. Thus, the PCL nanoparticle proposed here is an innovative nanomedicine for melanoma therapy and represents a strong candidate for specific pre-clinical and in vivo studie

A rapid and sensitive High-Performance Liquid Chromatography method with fluorescence detection for quantification of melatonin in small volume rat plasma samples: application to a preclinical study to determine the oral pharmacokinetics of melatonin under gestational conditions

Abstract A novel, simple and sensitive high-performance liquid chromatography with fluorescence detection method was developed and validated for the characterization of the preclinical pharmacokinetics of melatonin under pregnant conditions. Plasma samples (25 µL) were treated with 30 µL of ethanol absolute (containing the internal standard, IS). After a centrifugation process, aliquots of supernant (5 µL) were injected into the chromatographic system. Compounds were eluted on a Xbridge C18 (150 mm x 4.6 mm i.d., 5 µm particle size) maintained at 30°C. The mobile phase consisted in a mixture of aqueous solution of 0.4% phosphoric acid and acetonitrile (70:30 v/v). The wavelengths were set at 305 nm (excitation) and 408 nm (emission) and the total analysis time was 8 min/sample. All validation tests were obtained with accuracy and precision, according to FDA guidelines, over the concentration range of 0.005-20 µg/mL. Pharmacokinetic study showed that melatonin systemic exposure increased from day 14, with a significant difference at 19 days of gestation compared to the control group. Our findings suggest a decreased metabolism of melatonin as result of temporary physiological changes that occur throughout pregnancy. However, other maternal physiological changes cannot be ruled out.

Anti - proliferative and anti - inflammatory activity of triterpene extracts from plant species belonging to Lamiaceae family / Actividad antiproliferativa y antinflamatoria de los extractos de triterpenos de plantas pertenecientes a la familia Lamiaceae

Triterpenes are very important secondary metabolites with wide structural diversity and significant role in pharmacy and medicine. In the present research, a comparative study of pharamacological activities of the triterpene fractions obtained from several plant species belonging to Lamiaceae family, was carried out. In - vitro anti - proliferative activity was performed using a standard proliferation assay based on tetrazolium salts. In vitro anti - inflammatory activity of triterpene fractions was determined by an assay of inhibition of albumin denaturation. In general, the triterpene fractions obtained from plant species belonging to Lam iaceae family showed a strong anti - proliferative activity and anti - inflammatory activity. The triterpene fraction of Rosmarini folium showed the strongest anti - proliferative activity (GI 50 range from 4 to 37 µg/ml) and the strongest anti - inflammatory activ ity in the range from 57.27% to 80.69%. This comparative study provides scientific evidence to support the traditional use of Lamiacae plant species for medical purposes as anti - inflammatory and anti - proliferative medicines.

Los triterpenos son metabolitos secundarios muy importantes, con una amplia diversidad estructural y un rol significativo en la farmacia y la medicina. En esta investiga ción, se realizó un estudio comparativo de las actividades farmacológicas de las fracciones de triterpenos obtenidas de varias especies de plantas pertenecientes a la familia Lamiaceae. La actividad antiproliferativa in vitro se realizó mediante un ensayo estándar de proliferación basado en sales de tetrazolio. Se determinó la actividad antinflamatoria de las fracciones de triterpeno s mediante un ensayo de inhibición de desnaturalización de la albúmina. En general, las fracciones de triterpeno s obtenidas de las plantas pertenecientes a la familia Lamiaceae mostraron una actividad antiproliferativa y antinflamatoria fuerte. La fracción de triterpeno de Folium Rosmarini mostró la actividad antiproliferativa más fuerte (rango GI 50 entre 4 y 37 µg/m L ) y la más f uerte actividad antinflamatoria en el rango de 57,27% a 80,69%. Este estudio comparativo provée evidencia científica para apoyar el uso tradicional de especies de plantas Lamiaceae para usos médicos como medicinas antinflamatorios y antiproliferativas.

Effects of cinnamon oil and its main constituents, cinnamic acid and cinnamaldehyde, on 1 - methyl - 4 - phenyl - 1,2,3,6 - tetrahydropyridine - induced neurodegeneration in PC - 12 cells / Efectos del aceite de canela y sus principales componentes, ácido cinámico y cin amaldehído en la neurodegeneración inducida por 1 - metil - 4 - fenil - 1,2,3,6 - tetrahidropiridina en células PC 12

Cinnamon ( Cinnamomum verum J. Presl) is a well - known medicinal plant considered as an effective treatment for neurological disorders based on Persian medicine . The aim of the present study was assessing the effect of cinnamon oil, cinnamic acid, and cinnamaldehyde, on the in vitro model of Parkinson's disease (PD). Cinnamon oil, prepared in sesame oil, was phytochemically analyzed using high performance liquid chromatography (HPLC). Pheochromocytoma - 12 (PC - 12) cells were treated with 1 - methyl - 4 - phenyl - 1,2,3,6 - tetrahydropyridine (MPTP) as an in vitro model of neurodegeneration in PD. Cell viability, activity of caspase enzymes, and formation of reactive oxygen sp ecies (ROS) were evaluated. MPTP significantly decreased cell viability and increased Casp activity, as well as ROS formation. Cinnamon oil and cinnamic acid at 200 µg/m L could significantly reverse MPTP - induced abnormalities in PC - 12 cells including Casp activity and ROS formation. Our study supports the beneficial effect of cinnamon oil in neurodegeneration. Furt her investigations are needed to clarify the mechanisms and main active components.

La canela ( Cinnamomum verum J. Presl) es una planta medicinal m uy conocida, y considerada como un tratamiento efectivo para patologías neurológicas según la medicina persa. El objetivo de este estudio fue evaluar el efecto del aceite de canela, el ácido cinámico, y el cinamaldehído, en un modelo in vitro de la enferme dad de Parkinson (PD). El aceite de canela, preparado en aceite de sésamo, fue analizado fitoquímicamente usando cromatografía líquida de alta eficacia (HPLC). Se trataron células con feocromocitoma - 12 (P - 12) usando 1 - metil - 4 - fenil - 1,2,3,6 - tetrahidropiridi na (MPTP) como un modelo in vitro de neurodegeneración en PD. Se evaluó la viabilidad celular, actividad de enzimas caspasa, y formación de especies reactivas del oxígeno (ROS). El tratamiento con MPTP disminuyó significativamente la viabilidad celular y a umentó la actividad casp, así la formación de ROS. Aceite de canela y ácido cinámico a 200 µg/mL podría revertir significativamente las anormalidades inducidas por MPTP en células PC - 12, incluyendo la actividad casp y la formación de ROS. Nuestro estudio e ntrega sustento sobre los efectos benéficos del aceite de canela en la neurodegeneración. Se requiere más investigación para clarificar los mecanismos y los principales componentes activos.

Development, characterization and anti - Mycobacterium tuberculosis activity of solid lipid nanoparticles for oral Piper corcovadensis roots extract delivery / Desarrollo, caracterización y actividad anti - Mycobacterium tuberculosis de nanopartículas lipídicas sólidas para la administración oral de extracto de raíces de Piper corcovadensis

The present study thus aimed at the development and physicochemical characterization of solid lipid nanoparticles loaded with crude extract of Piper corcovadensis roots (SLN - CEPc) and chitosan - coated solid lipid nanoparticles loaded with crude extract of P. corcovadensis roots (C - SLN - CEPc), as well as the determination of its antimycobacterial activity against Mycobacterium tuberculosis H37Rv, its cytotoxicity against the Vero cell line and evaluation in the hemolysis assay. Both formulat ions containing the encapsulated extract showed high encapsulation efficiency, formed by a monodispersed system with small and spherical particles, and there was no aggregation of particles. In the biological assays, SLN - CEPc and C - SLN - CEPc showed promisin g anti - M. tuberculosis activity with a minimum inhibitory concentration (MIC) of 12.5 µg/mL, whereas the cytotoxic concentrations obtained at 50% (CC 50 ) in Vero cells were 60.0 and 70.0 µg/mL, respectively. Therefore, nanoencapsulation showed satisfactory results, justifying its usage in the development of new products.

El presente estudio apuntó al desarrollo y caracterización fisicoquímica de na nopartículas lípidas en estado sólido, cargadas con extracto crudo de raíz de Piper c orcovadensis (SLN - CEPc) y nanopartículas lípidas en estado sólido cubiertas con quitosano cargadas co n extracto crudo de raíz de P. corcovadensis (C - SLN - CEPc), así como la determinación de su actividad antimico bacterial contra Mycobacterium tuberculosis H37Rv, su citotoxicidad contra la línea celular Vero y su evaluación en ensayo de hemólisis. Ambas formulaciones que contenían el extracto encapsulado mostraron alta eficien cia de encapsulación, formado por un sistema monodispersado con pequeñas partículas esféricas, y no hubo agregación de partículas. En los ensayos biológicos, SLN - CEPc y C - SLN - CEPc mostraron un a prometedora actividad anti - M. tuberculosis con una mínima conc entración inhibitoria (MIC) de 12,5 µg/mL, mientras que las concentraciones citotóxicas obtenidas al 50% (CC 50 ) en células Vero estuvo en 60,0 y 70,0 µg/mL, respectivamente. Por lo tanto, la nanoencapsulación mostró resultados satisfactorios, justificando su uso en el desarrollo de nuevos productos.

Assessment of aflatoxin B1 contamination in rice and maize / Evaluación de la contaminación por aflatoxina B1 en arroz y maíz

Introduction: Aflatoxins B1 are among the most common poisonous mycotoxins produced by certain fungi that harm animals and crops. Mycotoxins can cause a variety of adverse health effects and pose a serious health threat to humans. The Maximum Residue Limits of aflatoxin B1 in processed cereals and ingredients are 2 parts per billion (ppb) and 5 ppb, respectively. Objectives: To evaluate the status of aflatoxin B1 contamination in rice, corn and staple food produced in Ha Giang province compared with the maximum permitted levels. Methods: A total of 210 rice and maize samples were analyzed to quantify the level of aflatoxin B1. Analysis of mycotoxins was conducted by High Performance Liquid Chromatography using a fluorescence detector. Results: It was found that rice, rice products, maize, and maize products had a mean aflatoxin B1 content of 1.79 ppb, 2.55 ppb, 2.19 ppb, and 6.35 ppb, respectively. The results also showed that 71.9 percent of samples were contaminated with mycotoxins, and 14.28 percent of samples exceeded the maximum allowable limit. Conclusion: The concentration of aflatoxin B1 in 14.28 percent of the samples are over permissible limits by nationwide regulations (AU)

Introducción: La aflatoxina B1 se encuentra entre las micotoxinas más comunes y venenosas producidas por ciertos hongos que dañan a los animales y los cultivos. Las micotoxinas pueden causar una variedad de efectos adversos para la salud y representar una grave amenaza para la salud de los seres humanos. Los límites máximos de residuos de aflatoxina B1en cereales e ingredientes procesados son de 2 ppb y 5 ppb, respectivamente. Objetivos: Evaluar el estado de contaminación por aflatoxina B1 en arroz, maíz y alimentos básicos producidos en la provincia de Ha Giang, en comparación con los niveles máximos permitidos. Métodos: Se analizaron un total de 210 muestras de arroz y maíz para cuantificar el nivel de aflatoxina B1. El análisis de micotoxinas se realizó mediante cromatografía líquida de alta resolución, utilizando un detector de fluorescencia. Resultados: Se encontró que el arroz, los productos de arroz, el maíz y los productos de maíz tenían un contenido medio de aflatoxin B1, de 1,79 ppb, 2,55 ppb, 2,19 ppb y 6,35 ppb, respectivamente. Los resultados también mostraron que el 71,9 por ciento de las muestras estaban contaminadas con micotoxinas y el 14,28 por ciento de las muestras excedieron el límite máximo permitido. Conclusión: La concentración de aflatoxina B1 en el 14,28 por ciento de las muestras está por encima de los límites permisibles por la norma nacional(AU)

Desarrollo inicial de un fitofármaco derivado del jugo de citrus limón (limón) para el tratamiento de crisis hipertensivas / Development of a phyto-drug derived from citrus lemon juice to treat acute blood pressure elevations

Objetivo: Obtener un fitofármaco a base de extracto seco de limón con estándares de calidad, procesamiento amigable con el medio ambiente y con antecedentes de efectividad y seguridad, evaluando sus efectos en pacientes hipertensos con frecuentes crisis hipertensivas. Métodos y resultados: Se seleccionó la población de limónes de las Regiones Metropolitana y Coquimbo con las concentraciones más altas de ácido ascórbico y flavonoides totales, respectivamente. Se utilizó liofilización para obtener jugo de limón en polvo. Sólo durante el período de estudio, se informó un caso de aumento/ crisis hipertensiva. La formulación con la concentración más alta de ácido ascórbico disminuyó la presión arterial sistólica y diastólica en 16 mmHg desde 10 minutos hasta 60 minutos. Por otro lado, la formulación con mayor concentración de flavonoides disminuyó la presión arterial en 12 a 30 mmHg desde 5 a 60 minutos. Conclusión: Se obtuvo un producto innovador como complemento al manejo de los aumentos de presión arterial. Los principios activos con mayor contribución al mecanismo antihipertensivo del jugo de limón corresponden a compuestos fenólicos, específicamente, flavonoides.

Aim: To obtain a product based on a dry extract of standardized phytodrug lemon with high quality standards, using an environmentally friendly process; to evaluate its effects in hypertensive patients suffering frequent episodes of hypertensive crisis. Methods and Results: Lemons with high concentration of ascorbic acid (Coquimbo Region) or total flavonoids (Metropolitan Region) were selected. Lyophilization was used to obtain juice powder. During the study period only one case of hypertensive crisis was reported. The formula with the highest concentration of ascorbic acid decreased systolic and diastolic blood pressure by 16 mmHg from 10 minutes up to 60 minutes. On the other hand, the formula with the highest concentration of flavonoids decreased blood pressure from 12 to 30 mmHg between 5 and 60 minutes. Conclusion: An novel product to help decrease acutely elevated blood pressure was obtained that can be used as a complement to the management of acute blood pressure rises. The active principles with greater contribution to the antihypertensive mechanism of lemon juice corresponded to phenolic compounds, specifically flavonoids.

Chemical components of Magnoliae Officinalis Cortex of different origins and with different tree ages before and after being processed with ginger juice:a qualitative and quantitative analysis / 中国中药杂志

This study aimed to investigate the impact of ginger juice on chemical profile of Magnoliae Officinalis Cortex(MOC) when they were processed together. Ultra-high-performance liquid chromatography coupled to quadrupole-orbitrap high-resolution mass spectrometry(UHPLC-Q-Orbitrap HRMS) was used for qualitative analysis of the chemical component of MOC samples before and after being processed with ginger juice. UPLC was performed to observe the content variation of eight main components in processed MOC. A total of 174 compounds were identified or tentatively deduced from processed and unprocessed MOC samples according to MS data obtained in positive and negative ion mode. After MOC was processed with ginger juice, the peak areas of most phenolics increased, while the peak areas of most phenylethanoid glycosides decreased; as for neolignans, oxyneolignans, other lignans and alkaloids, changes in the peak area were variable, and the peak areas of terpenoid-lignans varied little. Additionally, gingerols and diarylheptanoids were only detected in the processed MOC sample. The contents of syringin, magnoloside A, and magnoloside B decreased significantly in the processed MOC sample while no significant difference was observed in the contents of magnoflorine, magnocurarine, honokiol, obovatol, and magnolol. This study comprehensively explored the content variation of chemical components in processed and unprocessed MOC samples derived from different regions and with different tree ages using UPLC and UHPLC-Q-Orbitrap HRMS, and summarized the variation characteristics of various compounds. The results provide a data foundation for further research on pharmacodynamic substances of MOC processed with ginger juice.

Comparison of HPLC fingerprints and determination of main components of Viticis Fructus from different species / 中国中药杂志

In order to comprehensively evaluate the quality of Viticis Fructus, this study established HPLC fingerprints and evaluated the quality of 24 batches of Viticis Fructus samples from different species by similarity evaluation and multivariate statistical analysis(PCA, HCA, PLS-DA). On this basis, an HPLC method was established to compare the content differences of the main components, including casticin, agnuside, homoorientin, and p-hydroxybenzoic acid. The analysis was performed on the chromatographic column(Waters Symmetry C_(18)) with a gradient mobile phase of acetonitrile(A)-0.05% phosphoric acid solution(B) at the flow rate of 1 mL·min~(-1) and detection wavelength of 258 nm. The column temperature was 30 ℃ and the injection volume was 10 μL. The HPLC fingerprint of 24 batches of Viticis Fructus samples was established with 21 common peaks, and nine peaks were identified. Similarity analysis was carried out based on chromatographic data of 24 batches of chromatographic data of Viticis Fructus, and the results showed that except for DYMJ-16, the similarity of Vitex trifolia var. simplicifolia was ≥0.900, while that of V. trifolia was ≤0.864. In addition, the similarity analysis of two different species showed that the similarity of 16 batches of V. trifolia var. simplicifolia was 0.894-0.997 and that of the eight batches of V. trifolia was between 0.990 and 0.997. The results showed that the similarity of fingerprints of these two species was different, but the similarity between the same species was good. The results of the three multivariate statistical analyses were consistent, which could distinguish the two different species. The VIP analysis results of PLS-DA showed that casticin and agnuside contributed the most to the distinction. The content determination results showed that there was no significant difference in the content of homoorientin and p-hydroxybenzoic acid in Viticis Fructus from different species, but the content of casticin and agnuside was significantly different in different species(P<0.01). The content of casticin was higher in V. trifolia var. simplicifolia, while agnuside was higher in V. trifolia. The findings of this study show that there are differences in fingerprint similarity and component content of Viticis Fructus from different species, which can provide references for the in-depth study of the quality and clinical application of Viticis Fructus.

Metabolomics study of Berberidis Radix in intervening ulcerative colitis based on UPLC-Q-TOF-MS / 中国中药杂志

The effect of Tujia medicine Berberidis Radix on endogenous metabolites in the serum and feces of mice with ulcerative colitis(UC) induced by dextran sulfate sodium(DSS) was analyzed by metabolomics technology to explore the metabolic pathway and underlying mechanism of Berberidis Radix in the intervention of UC. The UC model was induced in mice by DSS. Body weight, disease activity index(DAI), and colon length were recorded. The levels of tumor necrosis factor-α(TNF-α) and interleukin-10(IL-10) in colon tissues were determined by ELISA. The levels of endogenous metabolites in the serum and feces were detected by ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were employed to characterize and screen differential metabolites. The potential metabolic pathways were analyzed by MetaboAnalyst 5.0. The results showed that Berberidis Radix could significantly improve the symptoms of UC mice and increase the level of the anti-inflammatory factor IL-10. A total of 56 and 43 differential metabolites were identified in the serum and feces, respectively, belonging to lipids, amino acids, fatty acids, etc. After the intervention by Berberidis Radix, the metabolic disorder gradually recovered. The involved metabolic pathways included biosynthesis of phenylalanine, tyrosine, and tryptophan, linoleic acid metabolism, phenylalanine metabolism, and glycerophospholipid metabolism. Berberidis Radix can alleviate the symptoms of mice with DSS-induced UC, and the mechanism may be closely related to the re-gulation of lipid metabolism, amino acid metabolism, and energy metabolism.

Application of high-speed counter current chromatography in extraction and separation of alkaloids in natural products / 中国中药杂志

Alkaloids, widespread in plants, have a series of pharmacological activities and have been widely used to treat various diseases. Because alkaloids are usually presented in multicomponent mixtures and are deeply low in content, they are very difficult to extract and separate by traditional methods. High-speed counter current chromatography(HSCCC) is a kind of liquid-liquid chromatography without solid support phase, which has the advantages of large injection volume, low cost, and no irreversible adsorption. Compared with the traditional methods of extraction and separation of alkaloids, HSCCC can ensure the separation of many different alkaloids at one time, with a high recovery and large amount. In this paper, the advantages and disadvantages of HSCCC compared with traditional separation methods were discussed and the solvent system and elution mode of HSCCC used to separate alkaloids in recent years were summarized by referring to the relevant literature to provide some references for the separation of alkaloids by HSCCC.

Separation, characterization and anti-psoriasis effect of self-assembled nanoparticles from Shaoyao Gancao Decoction / 中国中药杂志

This study aims to separate and characterize self-assembled nanoparticles(SAN) from Shaoyao Gancao Decoction(SGD) and determine the content of active compounds. Further, we aimed to observe the therapeutic effect of SGD-SAN on imiquimod-induced psoriasis in mice. The separation of SGD was performed by dialysis, and the separation process was optimized by single factor experiment. The SGD-SAN isolated under the optimal process was characterized, and the content of gallic acid, albiflorin, paeoniflorin, liquiritin, isoliquiritin apioside, isoliquiritin, and glycyrrhizic acid in each part of SGD was determined by HPLC. In the animal experiment, mice were assigned into a normal group, a model group, a methotrexate group(0.001 g·kg~(-1)), and SGD, SGD sediment, SGD dialysate, and SGD-SAN groups of different doses(1, 2, and 4 g·kg~(-1)) respectively. The psoriasis grade of mice was evaluated based on the pathological changes of skin lesions, the content of inflammatory cytokines, organ index and other indicators. The results showed that SAN obtained by centrifugation at 13 000 r·min~(-1) for 30 min was stable after dialysis for 4 times, which were uniform spherical nanoparticles with the particle size of(164.43±1.34) nm, the polydispersity index of(0.28±0.05), and the Zeta potential of(-12.35±0.80) mV. The active compound content accounted for more than 70% of SGD. Compared with the model group, SAN and SGD decreased the skin lesion score, spleen index, and inflammatory cytokine levels(P<0.05 or P<0.01) and alleviated the skin thickening and infiltration of inflammatory cells. However, the sediment group and the dialysate group had no obvious effect. SGD showed a good therapeutic effect on imiquimod-induced psoriasis in mice, and SAN demonstrated the effect equivalent to SGD in a dose-dependent manner. Therefore, we conclude that the SAN formed during decocting is the main active form of SGD, which can lower the levels of inflammatory cytokines, promote the normal differentiation of keratinocytes, and reduce the infiltration of inflammatory cells in the treatment of psoriasis lesions in mice.

Component identification and analysis in vivo of Sanhan Huashi formula / 中国中药杂志

Sanhan Huashi formula(SHF) is the intermediate of a newly approved traditional Chinese medicine(TCM) Sanhan Huashi Granules for the treatment of COVID-19 infection. The chemical composition of SHF is complex since it contains 20 single herbal medicines. In this study, UHPLC-Orbitrap Exploris 240 was used to identify the chemical components in SHF and in rat plasma, lung and feces after oral administration of SHF, and heat map was plotted for characterizing the distribution of the chemical components. Chromatographic separation was conducted on a Waters ACQUITY UPLC BEH C_(18)(2.1 mm×100 mm, 1.7 μm) using 0.1% formic acid(A)-acetonitrile(B) as mobile phases in a gradient elution. Electrospray ionization(ESI) source was used to acquire data in positive and negative mode. By reference to quasi-molecular ions and MS/MS fragment ions and in combination with MS spectra of reference substances and compound information in literature reports, 80 components were identified in SHF, including 14 flavonoids, 13 coumarins, 5 lignans, 12 amino-compounds, 6 terpenes and 30 other compounds; 40 chemical components were identified in rat plasma, 27 in lung and 56 in feces. Component identification and characterization of SHF in vitro and in vivo lay foundations for disclosure of its pharmacodynamic substances and elucidation of the scientific connotation.

Nature-effect transformation mechanism of mulberry leaves and silkworm droppings based on chemical composition analysis / 中国中药杂志

Starting with the relationship between mulberry leaves and silkworm droppings as food and metabolites, this study systematically compared the chemical components, screened out differential components, and quantitatively analyzed the main differential components based on ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) and UPLC-Q-TRAP-MS combined with principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA). Moreover, the in vitro enzymatic transformation of the representative differential components was studied. The results showed that(1) 95 components were identified from mulberry leaves and silkworm droppings, among which 27 components only exist in mulberry leaves and 8 components in silkworm droppings. The main differential components were flavonoid glycosides and chlorogenic acids.(2) Nineteen components with significant difference were quantitatively analyzed, and the components with significant differences and high content were neochlorogenic acid, chlorogenic acid, and rutin.(3) The crude protease in the mid-gut of silkworm significantly metabolized neochlorogenic acid and chlorogenic acid, which may be an important reason for the efficacy change in mulberry leaves and silkworm droppings. This study lays a scientific foundation for the development, utilization, and quality control of mulberry leaves and silkworm droppings. It provides references for clarifying the possible material basis and mechanism of the pungent-cool and dispersing nature of mulberry leaves transforming into the pungent-warm and dampness-resolving nature of silkworm droppings, and offers a new idea for the study of nature-effect transformation mechanism of traditional Chinese medicine.

Quality evaluation and multi-spectral identification of origin herbs of Lonicerae Japonicae Flos based on grey correlation-TOPSIS method / 中国中药杂志

The grey correlation-TOPSIS method was used to evaluate the quality of the origin herbs of Lonicerae Japonicae Flos, and the Fourier transform near-infrared(NIR) and mid-infrared(MIR) spectroscopy was applied to establish the identification model of origin herbs of Lonicerae Japonicae Flos by combining chemometrics and spectral fusion strategies. The content of neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, caffeic acid, secoxyloganin, isoquercitrin, isochlorogenic acid B, isochlorogenic acid A, and isochlorogenic acid C in six origin herbs of Lonicerae Japonicae Flos was determined by high-performance liquid chromatography(HPLC), and their quality was evaluated by the grey correlation-TOPSIS method. The Fourier transform NIR and MIR spectra of six origin herbs of Lonicerae Japonicae Flos(Lonicera japonica, L. macranthoides, L. hypoglauca, L. fulvotomentosa, L. confuse, and L. similis) were collected. At the same time, principal component analysis(PCA), support vector machine(SVM), and spectral data fusion technology were combined to determine the optimal identification method for the origin herbs of Lonicerae Japonicae Flos. There were differences in the quality of the origin herbs of Lonicerae Japonicae Flos. Specifically, there were significant differences between L. japonica and the other five origin herbs(P<0.01). The quality of L. similis was significantly different from that of L. fulvotomentosa, L. macranthoides, and L. hypoglauca(P=0.008, 0.027, 0.01), and there were also significant differences in the quality of L. hypoglauca and L. confuse(P=0.001). The PCA and SVM 2D models based on a single spectrum could not be used for the effective identification of the origin herbs of Lonicerae Japonicae Flos. The data fusion combined with the SVM model further improved the identification accuracy, and the identification accuracy of the mid-level data fusion reached 100%. Therefore, the grey correlation-TOPSIS method can be used to evaluate the quality of the origin herbs of Lonicerae Japonicae Flos. Based on the infrared spectral data fusion strategy and SVM chemometric model, it can accurately identify the origin herbs of Lonicerae Japonicae Flos, which can provide a new method for the origin identification of medicinal materials of Lonicerae Japonicae Flos.

Assay of amino acids in leaves of Eucommia ulmoides under arbor forest mode and leaf-oriented cultivation mode by pre-column derivatization HPLC / 中国中药杂志

This study aims to develop the pre-column derivatization high performance liquid chromatography(HPLC) method for the determination of 16 kinds of amino acids in Eucommia ulmoides leaves, and compare the content of amino acids in the leaves harvested at different time and under leaf-oriented cultivation mode(LCM) and arbor forest mode(AFM). The HPLC conditions are as below: phenyl isothiocyanate(PITC) as pre-column derivatization agent, Agilent ZORBAX C_(18 )column(4.6 mm×250 mm, 5 μm), mobile phase A of acetonitrile-water(80∶20), mobile phase B of 0.1 mol·L~(-1) sodium acetate solution-acetonitrile(94∶6), gradient elution, flow rate of 1.0 mL·min~(-1), injection volume of 5 μL, column temperature of 40 ℃, and detection wavelength of 254 nm. The HPLC profile indicated well separation of 16 kinds of amino acids and the amino acid content in E. ulmoides leaves was up to 16.26%. In addition, the amino acid content in leaves of E. ulmoides under LCM was higher than under AFM. The amino acid content varied with the harvesting time. Through orthogonal partial least squares discriminant analysis, the amino acids of E. ulmoides under LCM and AFM were compared, which can distinguish the leaves under LCM from those under AFM. Principal component analysis was applied to comprehensively score the amino acids of E. ulmoides leaves. The results showed that the score of leaves under LCM was higher than that under AFM. Nutritional evaluation results indicated that the proteins in E. ulmoides leaves belonged to high-quality vegetable proteins. The established method for the determination of amino acid content is reliable. With the amino acid content as index, the leaf quality of E. ulmoides under LCM is better than that under AFM. This study lays a theoretical basis for the promotion of LCM for E. ulmoides and the development of medicinal and edible products from E. ulmoides leaves.