Thermal and pH stability of Justicia spicigera (Mexican honeysuckle) pigments: Application of mathematical probabilistic models to predict pigments stability.

Kinetic and probabilistic (Time-to-Failure, TTF) models were used to predict the color (L*, a*, b* total color differences (ΔE), Hue and Chroma) stability of Justicia spicigera leaves pigments subjected to different temperatures (40 - 80 °C) and pHs (2 - 12). The change in pH caused different hues (from 60° = orange red to 268° = deep-blue) due to the shift effect of anthocyanins in the extract. Temperatures higher than 60 °C increased the color degradation. High heat sensitivity was observed at pH 4 (Ea = 90.27) and 10 (Ea = 154.99 kJ/mol). The Time-to-Failure model for both ΔE and Hue describes the effect of pH and temperature in the J. spicigera extracts. High pHs and temperatures applied to the extracts increased the probability of showing ΔEs > 4 or Hue changes over 20 %. Nearby the neutral region of pH, pigments of J. spicigera were more stable. The TTF model might be a useful tool to describe and predict the behavior of pigments added to foods.

Methodology for dyes purification by preparative bidimensional offline reversed-phase high-performance liquid chromatography separation based on mobile phase pH change: Case study of Justicia spicigera.

The extensive characterization of the natural dyes involves the purification of their colored compounds for their structural analysis and stability studies. As most natural compounds being ionizable, herein is presented the optimization of an easy and affordable preparative bidimensional offline reversed-phase high-performance liquid chromatography purification based on mobile phase pH change. At the analytical scale, several combinations of stationary phases and mobile phases at different pH values were investigated first. The orthogonality between the dimensions was quantitatively evaluated using the nearest-neighbor distance approach. The optimized separation method was transferred to the preparative scale for the successful isolation of two colored compounds from Justicia spicigera, a traditional dye plant from Central America. They were identified as perisbivalvine B (2-amino-8-hydroxy-7-methoxy-3H-phenoxazin-3-one) and one of its derivatives, also found in another tinctorial plant species, Peristrophe bivalvis growing in Asia.

Microencapsulates by spray of Lacticaseibacillus rhamnosus GG from fermented whole or skimmed cow's milk added with Mexican honeysuckle (Justicia spicigera) extract using mesquite gum as carrier agent.

This work aimed to evaluate the effect of the addition 5°Bx Mexican honeysuckle (Justicia spicigera) extract (JSE) in spray dried encapsulates of whole and skimmed unfermented and fermented cow's milk with Lacticaseibacillus rhamnosus (LR). All samples were spray dried at 160 °C. Samples were analyzed in physical properties (moisture content, water activity (a w ), L∗, a∗, b∗, Hue, Chroma color parameters, particle size), LR content, and bioactive compounds (total anthocyanins (TA), total phenolic content (TPC), and antioxidant capacity (AC) using the DPPH assay). Results showed that the load of LR was in the range 6.79-7.44 Log10 (CFU/mL) cycles after fermentation, lower values were obtained when JSE was added before fermentation. In addition, LR remains after drying in fermented samples but decrease about 1 Log10 (CFU/mL) cycle. LR was 4.46 Log10 (CFU/mL) in the fermented skimmed milk-J. spicigera extract powder. All powders had a w and moisture content below 0.295 and 6.51%, respectively. Color of powders depended on the moment of addition of JSE and fermentation. Powders from fermented milk were pale brownish/orangey/red (Hue = 44.91-59.7) and unfermented and J. spicigera extract-maltodextrin solution (12% w/v) powders were purple (Hue = 314.52-326.68). Higher particle sizes (52.3-104.7 µm) were obtained with whole milk fermented and unfermented powders. On the contrary, skimmed milk and JSE without milk protein had values in the range 15.56-44.0 µm. TPC in powders were higher (16.96-33.81 mg GAE/g powder db) compared with TA (0.27-0.64 mg Peonidin-3,5-diglucoside/g powder db). TPC increased with fermentation and remain after spray drying. The AC and TPCs were highly correlated and had antioxidant capacity of 10.18 mg TE/g powder db. The principal component analysis showed that the type of milk and fermentation separate the powders in four groups, depending on their physical and antioxidant properties. Encapsulated pigments could be used in formulations in the food industry to increase bioactive compounds and pigments in foods.

Antilipidemic and Hepatoprotective Effects of Ethanol Extract of Justicia spicigera in Streptozotocin Diabetic Rats.

Oxidative stress is a factor that contributes to the development of complications in diabetes; however, its effects can be counteracted using exogenous antioxidants that are found in some plants, which is why people turn to traditional medicines in the search for therapeutic treatment. Justicia spicigera has been demonstrated to have the capacity to reduce glycemic levels; however, its effects on non-insulin-dependent organs such as the liver have not been reported. During 30 days of administration of Justicia spicigera ethanol extract, the blood glucose and weight of rats were measured every 5 days. Once the treatment was concluded, the rats were sacrificed. Corporal weight, blood glucose, cholesterol, very-low-density lipoprotein (VLDL), triglycerides, total lipids, and liver profile were reduced in the diabetic condition and normalized with the application of ethanol extract from J. spicigera (EJS). Additionally, there was a significant increase in catalase and superoxide dismutase activity in the control diabetic rats, a decrease in their activity with the extract administration, and no effect on normoglycemic rats. In conclusion, EJS is considered to be capable of reducing oxidative stress by maintaining diminished lipid and liver function profiles in male Wistar rats with streptozotocin-induced diabetes.

Extraction and Microencapsulation of Bioactive Compounds from Muicle (Justicia spicigera) and Their Use in the Formulation of Functional Foods.

Bioactive compounds (BC) present in muicle leaves were extracted using the best extraction conditions obtained with a Box-Behnken experimental design, extracting 95% of BC. Microencapsulation of muicle BC was carried out by spray drying using DE10 maltodextrin (MD) and soy protein isolate (SPI) as encapsulating agents. The best conditions for the ethanolic extraction of BC from muicle were 30 °C, 40% aqueous ethanol, and one extraction for 1 h. The best spray drying encapsulating conditions for BC and antioxidant capacity (AC) using MD as an encapsulating agent were: 160-80 °C and 10% MD in the feeding solution, and for SPI: 180-70 °C and 5% SPI in the feeding solution. Microcapsules were added to yogurt and a sensory evaluation and retention of BC during 15-day storage at 4 °C was performed. Sensory evaluation showed that yogurt with added MD microcapsules had better acceptance than that with SPI microcapsules. Based on this, a jelly with added muicle MD microcapsules was also prepared which obtained better acceptance by the judges. At the end of the storage period, yogurt with SPI microcapsules showed better retention of BC and AC than yogurts with MD microcapsules; however, products with MD microcapsules had better acceptance.

Gastrointestinal activity of Justicia spicigera Schltdl. in experimental models.

Justicia spicigera Schltdl. (Acanthaceae) is used for treatment of gastrointestinal illnesses therapy in traditional medicine. The objective of this study was to give evidence of the antinociceptive and spasmolytic effects of the J. spicigera ethanol extract (JS EtOH) using in in vivo and/or in vitro assays. The JS EtOH exerted regulatory effect on the motility and a partial relaxing response on the intestinal tissue. Furthermore, a significant abdominal antinociceptive response was obtained in mice, which was totally abolished in the presence of 5-HT1A receptor antagonist (WAY100635, 0.1 mg/kg, s.c.) and partially by blocking opioid receptors (NX, 1 mg/kg, i.p.), whereas the inhibition of the NO synthesis (L-NAME, 30 mg/kg, i.p.) facilitated antinociception of this extract. Kaempferitrin was isolated and identified as major secondary metabolite. These results support the analgesic and spasmolytic-like activity of J. spicigera aerial parts involving inhibitory neurotransmission reinforcing the potential of this medicinal plant for alleviating pain.

Nanopartículas de plata de Justicia spicigera y sus potencialidades antimicrobianas en el control biológico de bacterias transmitidas por alimentos y hongos fitopatógenos / Silver nanoparticles from Justicia spicigera and their antimicrobial potentialities in the biocontrol of foodborne bacteria and phytopathogenic fungi

In the present work, the biosynthesis of silver-nanoparticles (AgNP) was evaluated using the aqueous extract from Justicia spicigera. The obtained silver nanoparticles were characterized using UV-visible spectroscopic techniques, energy dispersive X-ray spectrometers (EDS), zeta potential and dynamic light scattering. The antimicrobial activity of biosynthesized AgNP was tested against foodborne bacteria (Bacillus cereus, Klebsiella pneumoniae and Enterobacter aerogenes) and phytopathogenic fungi (Colletotrichum sp., Fusarium solani, Alternaria alternata and Macrophomina phaseolina). The elemental profile of synthesized nanoparticles using J. spicigera shows higher counts at 3 keV due to silver, confirming the formation of silver nanoparticles. Scanning electron microscopy (SEM) analysis showed a particle size between 86 and 100 nm with spherical morphology. AgNP showed effective antibacterial and antifungal activity against the tested organisms principally with B. cereus, K. pneumoniae, E. aerogenes, A. alternata and M. phaseolina. Therefore, further studies are needed to confirm the potential of AgNP from J. spicigera in the control of indicator organisms under field conditions.

En el presente trabajo se evaluó la biosíntesis de nanopartículas de plata (AgNP) en presencia de una sal de plata y extractos acuosos de Justicia spicigera. Las nanopartículas así obtenidas fueron caracterizadas mediante técnicas espectroscópicas UV-visibles, espectrómetros de rayos X de energía dispersiva (EDS), potencial zeta y dispersión de luz dinámica. La actividad antimicrobiana de las AgNP biosintetizadas se probó frente a diversas bacterias transmitidas por alimentos (Bacillus cereus, Klebsiella pneumoniae y Enterobacter aerogenes) y hongos fitopatógenos (Colletotrichum sp., Fusarium solani, Alternaria alternata y Macrophomina phaseolina). El perfil elemental de las nanopartículas sintetizadas utilizando el extracto de J. spicigera mostró valores altos a 3 keV, lo que confirma la formación de nanopartículas de plata. El análisis por microscopía electrónica de barrido (SEM) reveló un tamaño de partícula entre 86 y 100 nm, con morfología esférica. Las AgNP mostraron una actividad antibacteriana y antifúngica efectiva contra los organismos evaluados, principalmente contra B. cereus, K. pneumoniae, E. aerogenes, A. alternata y M. phaseolina. Se necesitan más estudios para confirmar el potencial de las AgNP derivadas de J. spicigera en el control de organismos indicadores en condiciones de campo.

Anxiolytic-like effect of the aqueous extract of Justicia spicigera leaves on female rats: A comparison to diazepam.

BACKGROUND: In Mexico, an infusion made from muicle (Justicia spicigera, Mexican honeysuckle) is used to treat menstrual problems and soothe people's "nerves" (i.e., anxiety and/or depression). However, no studies have yet substantiated these affects. Anxiety currently ranks as the seventh most common cause of disability worldwide, a fact that has intensified the search for new treatments with fewer adverse effects and better therapeutic efficacy. OBJECTIVE: The anxiolytic-like effect of the aqueous extract of muicle leaves (AEML) was analyzed, and the influence of the estrous cycle on its pharmacological effect in rats was explored. METHODS: Female Wistar rats were subjected to the elevated plus maze test to evaluate the anxiolytic effect, followed by an assessment of locomotor activity using the open field test. Diazepam was the reference drug used. Preliminary phytochemical tests were performed. RESULTS: The dose of 12  mg/kg of AEML increased the time to first entry, and the frequency of entries into, the open arms in the proestrus-estrus (P-E) and metestrus-diestrus phases (M-D), while also reducing the anxiety index in the P-E phases. These effects are similar to those observed with diazepam on the elevated plus maze test. None of the treatments showed effects on locomotor activity. The presence of flavonoids, sterols and terpenes in the AEML was confirmed. CONCLUSION: The anxiolytic-like effect of the dose of 12  mg/kg of AEML in the P-E phases could be due to the presence of the metabolites identified; i.e., flavonoids, sterols and/or terpenes.

Influence of Monometallic and Bimetallic Phytonanoparticles on Physiological Status of Mezquite.

The present study was conducted to evaluate the impact of monometallic and bimetallic nanoparticles (NPs) of copper (Cu) and silver (Ag) from Justicia spicigera on the photochemical efficiency and phenol pattern of Prosopis glandulosa. In this study, the existence of localized surface plasmon resonance absorption associated with the nano-sized nature of Ag, Cu and Cu/Ag particles was confirmed by the presence of a single peak around 487, 585, and 487/580 nm respectively. Zeta potential and electrophoretic mobility were found to be 0.2 mV and 0.02 µmcm/(Vs) for synthesized NPs indicating less stability and thus tendency to agglomerate, and broad distribution of particles. Cu-NPs and Cu/Ag-NPs demonstrate that the dispersed phase is stable and has a minimum particle size at zeta potentials above -30 mV. Changes in phenolic compounds, total chlorophyll, and photochemical efficiency in leaves exposed to Ag, Cu and Cu/Ag phyto-nanoparticles were evaluated up to 72 hours. The results revealed that Ag-NP and Cu-NP from J. spicigera at 100 mg/L showed significant reduction in chlorophyll, epidermal polyphenol content and photochemical efficiency of P. glandulosa. In contrast, the application of bimetallic Cu/Ag-NP from J. spicigera showed a positive impact on physiological parameters of P. glandulosa after 72 h of exposure.

Silver nanoparticles from Justicia spicigera and their antimicrobial potentialities in the biocontrol of foodborne bacteria and phytopathogenic fungi.

In the present work, the biosynthesis of silver-nanoparticles (AgNP) was evaluated using the aqueous extract from Justicia spicigera. The obtained silver nanoparticles were characterized using UV-visible spectroscopic techniques, energy dispersive X-ray spectrometers (EDS), zeta potential and dynamic light scattering. The antimicrobial activity of biosynthesized AgNP was tested against foodborne bacteria (Bacillus cereus, Klebsiella pneumoniae and Enterobacter aerogenes) and phytopathogenic fungi (Colletotrichum sp., Fusarium solani, Alternaria alternata and Macrophomina phaseolina). The elemental profile of synthesized nanoparticles using J. spicigera shows higher counts at 3keV due to silver, confirming the formation of silver nanoparticles. Scanning electron microscopy (SEM) analysis showed a particle size between 86 and 100nm with spherical morphology. AgNP showed effective antibacterial and antifungal activity against the tested organisms principally with B. cereus, K. pneumoniae, E. aerogenes, A. alternata and M. phaseolina. Therefore, further studies are needed to confirm the potential of AgNP from J. spicigera in the control of indicator organisms under field conditions.

Optimization of ultrasonic-assisted extraction of phenolic compounds from Justicia spicigera leaves.

A Box-Behnken design (Extraction-time, pulse-cycle, sonication-amplitude) was employed to extract phenolic compounds from Justicia spicigera leaves by ultrasonic-assisted extraction. The muicle leaves extracts were analyzed measuring total phenolic compounds and antioxidant capacity. According to response surface methodology the optimal conditions of ultrasonic-assisted extraction to obtain the highest soluble phenolic content were 2 min (extraction time) for 0.7 s (pulse cycle) at 55% of sonication amplitude. Under these optimal conditions, the total phenolic content was higher when was used ultrasonic-assisted extraction (54.02 mg/g) than stirring (46.46 mg/g) and thermal decoction (47.76 mg/g); however, the antioxidant capacity from J. spicigera extracts did not increase by ultrasonic-assisted extraction. The extracts or aqueous infusions from J. spicigera leaves are used for therapeutic proposes, therefore the ultrasonic-assisted extraction is a useful technology to improve the extraction of phytochemicals from J. spicigera leaves.

Exploratory studies of some Mexican medicinal plants: Cardiovascular effects in rats with and without hypertension.

BACKGROUND: Papaveraceae Argemone mexicana L., Burseraceae Bursera simaruba (L.) Sarg., Acanthaceae Justicia spicigera Schltdl. and Selaginellaceae Selaginella lepidophylla (Hook. & Grev.) Spring., have been used in Mexican traditional medicine to treat hypertension. The objective of this study was to further characterize the cardiovascular effects of the methanol extracts of such plants. METHODS: The medicinal plants were collected and taxonomically identified; the methanol extract of each explored plant were administrated to conscious and unconscious male Wistar rats with and without glucose-induced hypertension. The blood pressure (BP) and heart rate (HR) were evaluated before and after the extract administration. Vascular reactivity experiments were conducted in rat aortic rings obtained from rats with and without sugar-induced hypertension, a model widely used to study such effects with cardiovascular agents. RESULTS: After oral administration in normotensive conscious rats all tested extracts decreased the HR, such effect was only observed in hypertensive conscious rats after the administration of B. simaruba; only A. mexicana and B. simaruba decreased the BP after oral administration. All extracts administrated by intravenous injection diminished the mean arterial pressure. Dose-response curves to cumulative concentrations of all the extracts promote vascular relaxation in precontracted aortas from rats with and without sugar-induced hypertension. CONCLUSIONS: The present study indicated that B. simaruba is worthy of further investigation as a potential phytotherapeutic agent for treating hypertension.

Justicia spicigera Schltdl. and kaempferitrin as potential anticonvulsant natural products.

Justicia spicigera Schltdl. is a vegetal species traditionally used to control epilepsy, but scientific evidence is required to reinforce this activity. The aim of the study was to evaluate the anticonvulsant-like activity of J. spicigera aqueous extract (JsAE) and a bioactive compound. JsAE was assessed in a dose-response manner (30, 100 and 1000mg/kg, i.p.) using the pentylenetetrazol (PTZ)-induced seizures and maximal electroshock seizure (MES) test in mice in comparison to ethosuximide (ETX, reference drug 100mg/kg, i.p.) or phenytoin (25mg/kg, i.p.), respectively. Then a significant dosage (1000mg/kg, i.p.) was chosen to examine electrographic activity (EEG) in rats. Treatment groups were compared to the vehicle and ETX in the convulsive behavior alone or simultaneous to EEG after PTZ-induced seizures (80 or 35mg/kg, i.p., mice or rats). Kaempferitrin (a flavonoid of JsAE) and ETX were administered via intracerebroventricular (i.c.v, 4th ventricle, 1µg/µL) and tested in the presence of PTZ in rats. Results confirmed that JsAE delayed the onset of seizures and reduced frequency of tonic convulsion and mortality in mice. JsAE or kaempferitrin also decreased the EEG spikes frequency and amplitude in a similar manner than EXT in rats. In conclusion, these preliminary data give evidence of the potential of J. spicigera as possible anticonvulsant as recommended in folk medicine for treating epilepsy, where kaempferitrin is suggested as a partial responsible bioactive compound.

Physicochemical and antioxidant characterization of Justicia spicigera.

Extracts with water:ethanol (100:0, 70:30, 50:50, 30:70, 0:100) solutions from fresh (F), just dried (JD), dried and stored for one year (DS) Justicia spicigera leaves were obtained using the stirring and ultrasound techniques. Extracts were analyzed in physicochemical and antioxidant characteristics. Identification of chemical compounds by gas chromatography-mass spectroscopy (GC-MS) was also performed. 2.14±0.91, 5.67±1.70, and 8.52±4.97g Gallic acid equivalents/100g dry weight (d.w.) of phenolic compounds were found, in average, for F, JD, and DS J. spicigera, respectively. 2.22±1.31, 2.58±2.11, and 8.48±3.78g Trolox equivalents/100g d.w. were detected with the ABTS method and 0.49±0.33, 1.23±0.87, and 0.88±0.94g with the DPPH method for F, JD and DS J. spicigera, respectively. Eucalyptol, phytol, and azulene were identified as the main compounds. J. spicigera showed colors (green-iridescent, green-yellow, or pink of different intensities) and antioxidant characteristics depending on the solvent concentration. Extracts could be used in the food and pharmaceutical industries.

Antinociceptive Activity of an Ethanol Extract of Justicia spicigera.

Preclinical Research The aim of the present study was to evaluate the antinociceptive and sedative activity of an ethanol extract of Justicia spicigera an evergreen used in Mexican traditional medicine for the relief of pain, wounds, fever and inflammation. At 200 mg/kg po, the maximum dose examined, the ethanol extract of J. spicigera (JSE) had analgesic activity in mice in the acetic acid writhing test, the second phase of the formalin test and the tail flick test that was similar in efficacy to the NSAID, naproxen (150 mg/kg po). JSE was inactive in the hot plate test and and the ketamine-induced sleeping time test; it had no sedative effects. These results show that the ethanol extract from the leaves of J. spicigera has antinociceptive effects in mice without inducing sedation. Drug Dev Res 77 : 180-186, 2016. © 2016 Wiley Periodicals, Inc.