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Influence of drying on the flavor quality of spearmint (Mentha spicata L.).

Spearmint (Mentha spicata L.) was dried using three different drying methods: oven-drying at 45 degrees C, air-drying at ambient temperature, and freeze-drying. The effect of the drying method on the volatile compounds and on the structural integrity and sensory characteristics of the spice was evaluated. The volatile components from fresh and dried spearmint samples were isolated by simultaneous distillation-extraction (SDE) and analyzed by gas chromatography-mass spectrometry (GC-MS). A total of 28 compounds were identified, carvone, limonene, and 1,8-cineole, in that order, being the main components in all of the samples. Oven-drying at 45 degrees C and air-drying at ambient temperature were the methods that produced the best results. An increase in monoterpenes was observed in all of the dried samples, except in the freeze-dried samples that underwent freezing at -198 degrees C. Freeze-drying resulted in substantial losses in oxygenated terpenes and sesquiterpenes. The effect of each drying method on leaf structure was observed by scanning electron microscopy. From a sensory standpoint, drying the spearmint brought about a decrease in herbaceous and floral notes together with an increase in minty odor.

Air↗

Attenuation of benzoyl peroxide-mediated cutaneous oxidative stress and hyperproliferative response by the prophylactic treatment of mice with spearmint (Mentha spicata).

The modulating effect of spearmint (Mentha spicata) on benzoyl peroxide-induced responses of tumor promotion in murine skin was investigated. Benzoyl peroxide (BPO) is an effective cutaneous tumor promoter acting through the generation of oxidative stress, induction of ornithine decarboxylase activity and by enhancing DNA synthesis. BPO treatment (20 mg/animal) increased cutaneous microsomal lipid peroxidation and hydrogen peroxide generation. The activity of cutaneous antioxidant enzymes, namely catalase, glutathione peroxidase, glutathione reductase and glutathione S-transferase, was decreased and the level of cutaneous glutathione was depleted. BPO treatment also induced the ornithine decarboxylase activity and enhanced the [3H]thymidine uptake in DNA synthesis in murine skin. Prophylactic treatment of mice with spearmint extract (10, 15 and 20 mg/kg) 1 hr before BPO treatment resulted in the diminution of BPO-mediated damage. The susceptibility of cutaneous microsomal membrane to lipid peroxidation and hydrogen peroxide generation was significantly reduced (P < 0.05 ). In addition, depleted levels of glutathione, inhibited activity of glutathione dependent and antioxidant enzymes were recovered to a significant level (P < 0.01, P < 0.05 and P < 0.01, respectively). Similarly, the elevated ornithine decarboxylase activity and enhanced thymidine uptake in DNA synthesis was inhibited significantly (P < 0.05 ) in a dose-dependent manner. The protective effect of spearmint was dose dependent in all parameters. The result suggests that spearmint is an effective chemopreventive agent that may suppress BPO-induced cutaneous oxidative stress, toxicity and hyperproliferative effects in the skin of mice.

Animals↗

Investigation of biochemical and histopathological effects of Mentha piperita Labiatae and Mentha spicata Labiatae on liver tissue in rats.

The plant Mentha piperita, or peppermint, is commonly used in the treatment of loss of appetite, common cold, bronchitis, sinusitis, fever, nausea and vomiting, and indigestion as a herbal agent. In this study, we aimed to investigate biochemical and histological effects of M. piperita Labiatae, growing in the Yenisar Bademli town of Isparta city, and Mentha spicata Labiatae, growing in the Anamas high plateau of the Yenisar Bademli town, on the rat liver tissue. Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. Rats were divided into four groups of 12 animals: Group I received no herbal tea (control group); Group II received 20 g/L M. piperita tea; Group III received 20 g/L M. spicata tea; and Group IV received 40 g/L M. spicata tea. Herbal teas were prepared daily and provided at all times to the rats during 30 days as drinking water. Liver function tests, including aspartate aminotransferase (AST/GOT) and alanine aminotransferase (ALT/GPT) activities were measured. To evaluate liver antioxidant defences, superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), catalase (CAT) and thiobarbituric acid reactive substance (TBARS) activities were determined in the homogenates of liver tissue. In addition, liver tissues were submitted for histopathologic examination. AST and ALT activities were increased in Group II, Group III and Group IV gradually when compared with the control group. The difference between Group II and the control group was not statistically significant (P > 0.016). Increases in AST and ALT activities of Group III and Group IV were statistically significant when compared with the control group. SOD, GSH-Px and CAT activities were increased in Group II when compared with the control group but the difference was not statistically significant (P > 0.016). However, SOD, GSH-Px activities and the TBARS level were significantly increased, and CAT activity was significantly decreased in Group III when compared with the control group. In Group IV, while SOD, GSH-Px and CAT activities were decreased, the TBARS level was increased as compared with the control group (P < 0.0016). Histopathological evaluation of experimental groups revealed a mild to severe degree of hepatic damage when compared to the control group. In Group II, there was only minimal hepatocytes degeneration. In Groups III and IV, there were granular or ballooning hepatocyte degeneration and necrosis, sinusoidal and central vein dilatation. It was concluded that lipid peroxidation and hepatic damage occurs after M. piperita and M. spicata administration in rat liver and the damage seems to be dose dependent.

Alanine Transaminase↗

The effect of Mentha spicata Labiatae on uterine tissue in rats.

The plant Mentha spicata, or peppermint, is commonly used in the treatment of loss of appetite, common cold, bronchitis, sinusitis, fever, nausea and vomiting, and indigestion as a herbal agent. In this study, we aimed to investigate the biochemical and histological effects of M. spicata Labiatae, growing on the Anamas high plateau of Yenisarbademli town, on rat uterine tissue. Twenty female Wistar albino rats weighing 160+/-10 g were used for this study. Rats were divided into two groups of ten animals: group I received no herbal tea (control group) and group II received 20 g/L M. spicata tea. Control group rats were given commercial drinking water (Hayat DANONESA water). Herbal tea was prepared daily and provided at all times to the rats over 30 days as drinking water. Plasma malondialdehyde (MDA) levels were determined. In addition, uterine tissues were submitted for histopathologic examination. MDA levels were increased in group II when compared with the control group. The difference between group II and the control group was statistically significant (P<0.01). In the M. spicata Labiatae-treated group, histopathological changes like apoptosis and diffuse eosinophil leucocyte infiltration in surface and stromal glandular epithelium were observed in both endometrium and endocervix. It was concluded that lipid peroxidation and uterine damage occurs after M. spicata administration in rat uterus. Despite the beneficial effects of M. spicata Labiatae in indigestion, we should also be aware of the toxic effects when it is not used in the recommended fashion, at the recommended dose.

Animals↗

[Study of chemical constituents in active parts of Mentha spicata III].

OBJECTIVE: To study the chemical constituents of Mentha spicata. METHOD: The chemical constituents were isolated by silica gel column chromatography, and identified by physical and chemical characters and spectroscopic analysis. RESULT: Compounds I - V were obtained and their structures were elucidated as protocatechuic aldehyde (I), protocatechuic acid (II), chrysoeriol (III), 5, 6-dihydroxy-7, 8, 3', 4'-tetramethoxyflavone (IV), nodifloretin (V). CONCLUSION: Compound I and II were first isolated from the genus Mentha. Compound Ill, IV and V were isolated from M. spicata for the first time.

Benzaldehydes↗

Effect of Mentha piperita (Labiatae) and Mentha spicata (Labiatae) on iron absorption in rats.

AIM: The effect of Mentha piperita (Labiatae) and Mentha spicata (Labiatae) teas, which contain different phenol compounds, on iron metabolism was studied. These teas grow in different areas of the city of Isparta, Turkey. These herbals were given to the rats in tea. METHODS: Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. The rats were divided into four groups of 12 animals: Group I received no herbal tea (control group); Group II received 20 g/L M. piperita tea; Group III received 20 g/L M. spicata tea; Group IV received 40 g/L M. spicata tea. Herbal teas were prepared daily and provided at all times to the rats over 30 days as drinking water. RESULTS: M. piperita tea caused a decrease in serum iron and ferritin levels (P < 0.05), and caused an increase in unsaturated iron-binding capacity (UIBC) (P < 0.01). M. spicata tea caused no significant change in serum iron, ferritin levels and UIBC (P > 0.05). CONCLUSION: Both herbal teas inhibited Fe absorption. Inhibition caused by M. spicata tea was dose dependent. Therefore, when drinking these teas, their effect should be considered, especially for children and anemic patients.

Animals↗

Purification of 4S-limonene synthase, a monoterpene cyclase from the glandular trichomes of peppermint (Mentha x piperita) and spearmint (Mentha spicata).

The p-menthane monoterpenes of the Mentha species are biosynthesized from geranyl pyrophosphate via the monocyclic olefin 4S-limonene. A monoterpene cyclase was isolated from both Mentha x piperita (peppermint) and Mentha spicata (spearmint) that catalyzes the cyclization of geranyl pyrophosphate to 4S-limonene. This enzyme, 4S-limonene synthase, was purified to apparent homogeneity by dye ligand, anion exchange, and hydrophobic interaction chromatography. Since the monoterpenes of Mentha are synthesized and secreted in modified epidermal hairs called glandular trichomes, an extract of isolated glandular trichome cells was used as the source of this enzyme. A combination of gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that purified 4S-limonene synthase had a native molecular weight of 56,000 and was monomeric. The principal product of the enzyme was enantiomerically pure (-)-4S-limonene, and a catalytic constant of 0.3/s was determined. The basic properties of 4S-limonene synthase from both M. x piperita and M. spicata are identical and, in general, are similar to those of other monoterpene, sesquiterpene, and diterpene cyclases isolated from microorganisms and higher plants.

Chromatography, Liquid↗

[A study on chemical constituents in the herb of Mentha spicata].

OBJECTIVE: To investigate the constituents of anti-inflammatory and hemostatic active sites of Mentha spicata. METHOD: Chemical constituents were separated by solvent method and chromatography and identified by physicochemical properties and spectroscopic methods. RESULT: The structures were identified as ursane I, 3-methoxy-4-methylbenzaldehyde II, veratric acid III, 5-hydroxy-3',4',6,7-tetramethoxyflavone IV, diosmetin(V), thymonin VI, daucosterol VII. CONCLUSION: Among them, compounds I, II, III were first obtained from M. genus, IV, VI were separated from M. spicta for the first time.

Flavonoids↗

Two new monoterpenoid glycosides from Mentha spicata L.

Two new monoterpenoid glycosides, spicatoside A and spicatoside B, were isolated from the whole herbs of Mentha spicata L. which have anti-inflammatory and hemostatic activities. Their structures have been determined on the basis of spectral and chemical analysis. They are (+)-5-[1-(beta-D-glucopyranosyloxymethyl)ethenyl]-2-methyl-2-cyclohexen-1-one (1), and (-)-5-[[2-(beta-D-glucopyranosyloxy)-1-hydroxy-l-methyl]ethyl]-2-methyl-2-cyclohexen-1-one (2).

Glycosides↗

Biochemical and Histochemical Localization of Monoterpene Biosynthesis in the Glandular Trichomes of Spearmint (Mentha spicata).

The primary monoterpene accumulated in the glandular trichomes of spearmint (Mentha spicata) is the ketone (-)-carvone which is formed by cyclization of the C(10) isoprenoid intermediate geranyl pyrophosphate to the olefin (-)-limonene, hydroxylation to (-)-trans-carveol and subsequent dehydrogenation. Selective extraction of the contents of the glandular trichomes indicated that essentially all of the cyclase and hydroxylase activities resided in these structures, whereas only about 30% of the carveol dehydrogenase was located here with the remainder located in the rest of the leaf. This distribution of carveol dehydrogenase activity was confirmed by histochemical methods. Electrophoretic analysis of the partially purified carveol dehydrogenase from extracts of both the glands and the leaves following gland removal indicated the presence of a unique carveol dehydrogenase species in the glandular trichomes, suggesting that the other dehydrogenase found throughout the leaf probably utilizes carveol only as an adventitious substrate. These results demonstrate that carvone biosynthesis takes place exclusively in the glandular trichomes in which this natural product accumulates.

Journal Article↗

Production and characterization of polyclonal antibodies in rabbits to 4S-limonene synthase from spearmint (Mentha spicata).

Limonene synthase, a monoterpene cyclase from the oil glands of spearmint (Mentha spicata) leaves that catalyzes the conversion of geranyl pyrophosphate to (-)-4S-limonene, was purified, and polyclonal antibodies were generated in rabbits against the sodium dodecyl sulfate-denatured protein. Immunoblotting analysis revealed that the antibodies were very specific for denatured limonene synthase from all Mentha species tested. However, no immunological cross-reactivity was observed with denatured limonene synthases from Valencia oranges (Citrus sinensis, Rutaceae) or wormseed (Chenopodium ambrosioides, Chenopodiaceae). Furthermore, the antibody preparation did not detectably cross-react with other monoterpene cyclases from related angiosperm species of the Lamiaceae, Asteraceae, and Umbellifereae, or from conifer species, and no cross-reactivity was demonstrated toward several sesquiterpene cyclases of higher plant and fungal origin. Although the antibody preparation was highly selective for denatured limonene cyclase from Mentha, the antibodies did not recognize the native protein in several different types of experiments. Nevertheless, specificity for the target enzyme was unambiguously demonstrated when the antibody preparation was shown to cross-react with the cyclase protein expressed in Escherichia coli that harbored the corresponding limonene synthase cDNA gene from M. spicata.

Animals↗

Monoterpene biosynthesis: specificity of the hydroxylations of (-)-limonene by enzyme preparations from peppermint (Mentha piperita), spearmint (Mentha spicata), and perilla (Perilla frutescens) leaves.

Microsomal preparations from the epidermal oil glands of Mentha piperita, Mentha spicata, and Perilla frutescens leaves catalyze the NADPH- and O2-dependent allylic hydroxylation of the monoterpene olefin (-)-limonene at C-3, C-6, and C-7, respectively, to produce the corresponding alcohols, (-)-trans-isopiperitenol, (-)-trans-carveol, and (-)-perillyl alcohol. These transformations are the key steps in the biosynthesis of oxygenated monoterpenes in the respective species, and the responsible enzyme systems meet most of the established criteria for cytochrome P450-dependent mixed function oxygenases. The reactions catalyzed are completely regiospecific and, while exhibiting only a modest degree of enantioselectivity, are highly specific for limonene as substrate. Of numerous monoterpene olefins tested, including several positional isomers of limonene, only the 8,9-dihydro analog served as an alternate substrate for ring (C-3 and C-6) hydroxylation, but not side chain (C-7) hydroxylation. In addition to the regiospecificity of the allylic hydroxylation, these enzymes are also readily distinguishable based on differential inhibition by substituted imidazoles.

Cyclohexenes↗

Influence of autoclaved fungal materials on spearmint (Mentha spicata L.) growth, morphogenesis, and secondary metabolism.

The influence of autoclaved fungal materials such as culture filtrate, freeze-dried mycelium (FDM), mycelium suspension, and spore suspension (SS) on the growth, morphogenesis, and carvone production of spearmint (Mentha spicata L.) plants was studied. Fungal materials were either applied as a drench or spray on the plants. Spearmint plants (cv. "294099") drenched with SS (1 x 10(8) spores/ml) of Trichoderma reesei showed no significant differences in leaf numbers, root numbers, or shoot numbers compared with nontreated controls. However, significantly higher fresh weights and carvone levels were observed in plants drenched with T. reesei SS compared with the untreated controls. Fungal materials derived from Aspergillus sp., Fusarium graminearum, F. sporotrichoides, Penicillium sp., P. acculeatum, Rhizopus oryzae, and T. reesei were sprayed on spearmint foliage. F. graminearum, F. sporotrichoides, or R. oryzae elicited no enhanced growth, morphogenesis, or secondary metabolism responses. The best growth and morphogenesis responses were obtained employing Aspergillus sp., Penicillium sp., or T. reesei foliar sprays. For example, spearmint cv. "557807" plants sprayed with 100 mg/l FDM T. reesei isolate NRRL 11460 C30 stimulated higher fresh weights (75%), shoot numbers (39%), leaf numbers (57%), and root numbers (108%) compared with untreated plants. This effect was not dose-dependent because similar growth and morphogenesis responses were obtained by testing 10, 100, or 1000 mg/l FDM concentrations. Carvone levels in fungal-treated foliar-sprayed plants were comparable to nontreated controls. However, total carvone levels per plant were higher in fungal-treated plants because of their increased fresh weight.

Animals↗

Antioxidant activity measured in different solvent fractions obtained from Mentha spicata Linn.: an analysis by ABTS*+ decolorization assay.

Antioxidant compounds are abundantly available in plants and play an important role in scavenging free radicals, thus providing protection to humans against oxidative DNA damage. Mentha spicata Linn., commonly called spearmint, belongs to the family lamiaceae. It was selected in the present study because Mentha extracts have antioxidant properties due to the presence of eugenol, caffeic acid, rosmarinic acid and alpha-tocopherol. Four solvent fractions (hexane, chloroform, ethyl acetate and water) of ethanolic extract of dried leaves powder of M. spicata were analyzed for total antioxidant activity (TAA) and relative antioxidant activity (RAA) and compared with standard antioxidants such as Quercetin, beta-carotene, L-ascorbic acid and glutathione using ABTS*+ decolorization assay (ABTS/Potassium persulphate). The antioxidant activity was assumed to be from the total phenolic content of the ethanolic extract. Total phenolics are found to be highest in ethyl acetate fraction (54 mg/g) and least in hexane fraction (13 mg/g) and more or less similar in water and chloroform fractions (30-32 mg/g). TAA is found to be less in hexane and chloroform fractions (<53% at 50 microg/ml) and highest in ethyl acetate (95% at 20 microg/ml) and water (84% at 30 microg/ml) fractions. The RAA of ethyl acetate fraction is 1.1 compared to quercetin (at 5 microM/ml), but greater when compared to beta-carotene (15 microM/ml), L-ascorbic acid (15 microM/ml) and glutathione (15 microM/ml). The RAAs with these antioxidants are in the range of 1.31 -1.6. The values of RAAs for water fraction also show similar trend and are in the range of 1.0-1.4. The antioxidant activities of the solvent factions are closely related to the content of total phenolics present in them.

Antioxidants↗

Purification and characterization of an NADPH-cytochrome P450 (cytochrome c) reductase from spearmint (Mentha spicata) glandular trichomes.

Solubilized NADPH-cytochrome c (P450) reductase was purified to homogeneity from an extract of spearmint (Mentha spicata) glandular trichomes by dye-ligand interaction chromatography on Matrex-Gel Red A and affinity chromatography on 2', 5'-adenosine diphosphate agarose. SDS-PAGE of the purified enzyme preparation revealed the presence of two similar proteins with masses of 82 kDa (major) and 77 kDa (minor) that crossreacted on immunoblot analysis with polyclonal antibodies directed against NADPH-cytochrome P450 reductase from Jerusalem artichoke and from mung bean. Complete immunoinhibition of reductase activity was observed with both types of polyclonal antibodies, while only partial inhibition of activity resulted using a family of monoclonal antibodies directed against the Jerusalem artichoke cytochrome P450 reductase. Inhibition of the spearmint oil gland cytochrome c reductase was also observed with the diphenyliodonium ion. The K(m) values for the cosubstrates NADPH and cytochrome c were 6.2 and 3.7 microM, respectively, and the pH optimum for activity was at 8.5. The NADPH-cytochrome c reductase reconstituted NADPH-dependent (-)-4S-limonene-6-hydroxylase activity in the presence of cytochrome P450, purified from the microsomal fraction of spearmint oil gland cells and dilauroyl phosphatidyl choline. These characteristics establish the identity of the purified enzyme as a NADPH-cytochrome P450 reductase.

Antibodies↗

Investigation of biochemical and histopathological effects of Mentha piperita L. and Mentha spicata L. on kidney tissue in rats.

Peppermint plants have been used as a herbal medicine for many conditions, including loss of appetite, common cold, bronchitis, sinusitis, fever, nausea, vomiting and indigestion. This study is aimed at investigating the biochemical and histological effects of Mentha piperita L., growing in the Yenisar Bademli town of Isparta City, and Mentha spicata L., growing on the Anamas high plateau of Isparta City, on rat kidney tissue. Forty-eight male Wistar albino rats weighing 200-250 g were used for this study. Animals were divided into four experimental groups, each with 12 rats, as follows: control group (group I); 20 g/L M. piperita tea (group II); 20 g/L M. spicata tea (group III); 40 g/L M. spicata tea (group IV). The control group rats were given commercial drinking water (Hayat DANONESA water). The tea for the other groups was prepared daily and provided at all times to the rats during 30 days as drinking water. Plasma urea and creatinine levels were determined, and the levels of thiobarbituric acid reactive substance (TBARS) and the activities of glutathione peroxidase (GSH-Px), catalase (CAT) and superoxide dismutase (SOD) were studied in the homogenates of kidney tissue. The levels of plasma urea and creatinine were increased significantly (P < 0.0033) in groups III and IV when compared with group I. The activities of SOD and GSH-Px were decreased significantly (P < 0.0033) in group IV when compared with group I. The activities of CAT were decreased significantly in groups III and IV (P < 0.033, P < 0.0033, respectively) when compared with group I. TBARS levels were increased significantly (P < 0.0033) in groups III and IV when compared with group I. In groups II, III and IV, hydropic degeneration of tubular epithelial cells, the epithelial cells with picnotic nuclei and eosinophilic cytoplasm, tubular dilatation and enlargements in Bowman capsules were observed histologically. However, in group II histopathological changes were more slight than in groups III and IV. In group IV, in addition to these changes, extremely hydropic degeneration of tubular epithelial cells, some atrophic tubules and glomerules, and focal mononuclear cell infiltrations in the kidney tissues of the rats were observed. In conclusion, the results indicate that M. piperita does not show nephrotoxicity but M. spicata presents markedly nephrotoxic changes in rats.

Animals↗

Analysis of Indian mint (Mentha spicata) for essential, trace and toxic elements and its antioxidant behaviour.

Mint, belonging to the genus Mentha in the family Labiatae (Lamiaceae) is pungent in taste with post digestive effects and hot potency. Ten samples of mint leaves, collected from four different locations in North-West parts of India (Roorkee, Dehradun, Baghpat and Uttarkashi) were analysed for seven minor (Al, Na, K, Ca, Cl, Mg, and P) and 20 trace (As, Au, Ba, Br, Co, Cr, Cs, Eu, Fe, Hf, Hg, La, Mn, Rb, Sb, Sc, Se, Sn, Th and Zn) elements by instrumental neutron activation analysis (INAA). Also Ni, Cu and Cd were determined by atomic absorption spectrophotometry (AAS). Samples along with reference materials (RMs) and synthetic primary standard were irradiated at approximately 10(13) n cm-2s-1 and its gamma-activity was measured using HPGe detector and MCA system. Most elements were found in widely varying amounts depending on the location, e.g. Na (0.21-0.86 mg/g), K (12.4-53.3 mg/g) and Ca (5.82-16.8 mg/g) whereas mean contents of other nutrient elements in mint were: Fe (108+/-22 microg/g), Mg (4.83+/-0.92 mg/g), Mn (53.5+/-9.6 microg/g), P (3.88+/-0.94 mg/g), Cu (16.9+/-1.8 microg/g), Zn (21.0+/-4.7 microg/g) and Se (0.18+/-0.03 microg/g). Further, DPPH free radical scavenging activity test in diethyl ether extract showed 100% activity at approximately 40 microg/L suggesting it to be antioxidant in accordance with literature reports.

Antioxidants↗