Antioxidant Activity and Phenolic Content of Different Parts of Lotus and Optimization of Extraction Condition using Response Surface Methodology

Nelumbo nucifera Gaertn. (Nymphaeaceae) is commonly called lotus and its leaves are widely been used as functional ingredients due to its antioxidant activity. For maximum efficacy, optimized extraction condition was established using response surface methodology. The high F-values, low p-values and insignificant p-value for lack-of-fit supported the fitness of the model and yielded the second-order polynomial regression for the antioxidant activity. The optimized extract was obtained by the extraction of 1 g of lotus leaves with 40 mL of 50% MeOH at 10.0℃, which exerted 70.1% antioxidant activity. Close correlation between phenolic content and antioxidant activity suggested phenolic compounds as active constituents of lotus leaves. In addition, comparison of different parts of lotus demonstrated the most potent antioxidant activity of flowers, followed by leaves and roots. Taken together, these results provide useful information about lotus leaves for the development as antioxidant ingredients. In addition, flowers and roots as well as leaves are suggested as good sources for antioxidant activity.

Efficient Isolation of Dihydrophaseic acid 3′-O-β-D-Glucopyranoside from Nelumbo nucifera Seeds Using High-performance Countercurrent Chromatography and Reverse-phased High-performance Liquid Chromatography

High-performance countercurrent chromatography (HPCCC) coupled with reversed-phase high-performance liquid chromatography (RP-HPLC) method was developed to isolate dihydrophaseic acid 3′-O-β-D-glucopyranoside (DHPAG) from the extract of Nelumbo nucifera seeds. Enriched DHPAG sample (2.3 g) was separated by HPCCC using ethyl acetate/n-butanol/water system (6:4:10, v/v/v, normal-phase mode, flow rate: 4.0 mL/min) to give 23.1 mg of DHPAG with purity of 88.7%. Further preparative RP-HPLC experiment gave pure DHPAG (16.3 mg, purity > 98%). The current study demonstrates that utilization of CCC method maximizes the isolation efficiency compared with that of solid-based conventional column chromatography.

Biopharmaceuticals classification system(BCS) of nuciferin in Nelumbinis Folium / 中国中药杂志

Nuciferin is the main active ingredients in Nelumbinis Folium， which was proved to have good hypolipidemic， antioxidative and anti-inflammatory bioactivities. Currently， pharmacokinetic studies of nuciferin showed different results based on different animal models. evaluation experiments were low-cost， stable and controllable. Biopharmaceutical classification system(BCS) was an effective and reliable simulation method to evaluate the bioavailability of oral drugs. It was a scientific framework for classifying drugs or active pharmaceutical ingredients(API) according to their solubility and impermeability . In the study， BCS was applied in an active ingredient in traditional Chinese medicine(TCM)， which was consisted of numerous chemical components. To study the equilibrium solubility of nuciferine， ideal solution model， Ape blat model and polynomial model were adopted. The permeability was measured based on partition coefficient(logP) and distribution coefficient(logD). Besides， apparent permeabilities of Caco-2 cells and murine intestine tissues were evaluated. Nuciferine was classified as BCSⅠ， since it had a good solubility and permeability in all methods under acidic conditions. However， in neutral and alkaline environments， nuciferine was classified as BCSⅣ by using everted intestinal sac. It indicated that the species of experimental animals has a significant influence on the absorption of nuciferine. This experiment can provide data support to the prediction in a complex environment(medicinal materials and absorbed parts). The application of BCS on TCM ingredients provided a new method to evaluate and screen out the druggability of TCM ingredients.

Anti-inflammatory effects of Nelumbo leaf extracts and identification of their metabolites

BACKGROUND/OBJECTIVES: Nelumbo leaves have been used in traditional medicine to treat bleeding, gastritis, hemorrhoids, and halitosis. However, their mechanisms have not been elucidated. MATERIALS/METHODS: The present study prepared two Nelumbo leaf extracts (NLEs) using water or 50% ethanol. Inflammatory response was induced with LPS treatment, and expression of pro-inflammatory mediators (inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-α (TNF-α), interleukin (IL)-1β, and IL-6 and nitric oxide (NO) and prostaglandin E₂ (PGE₂) productions were assessed. To determine the anti-inflammatory mechanism of NLEs, we measured nuclear factor-κB (NF-κB) activity. Major metabolites of NLEs were also analyzed and quantified. RESULTS: NLEs effectively reduced the expression and productions of pro-inflammatory mediators such as IL-1β, IL-6, TNF-α, PGE₂, and NO. NLEs also reduced NF-κB activity by inhibiting inhibitor of NF-κB phosphorylation. Both extracts contained catechin and quercetin, bioactive compounds of NLEs. CONCLUSIONS: In this study, we showed that NLEs could be used to inhibit NF-κB-mediated inflammatory responses. In addition, our data support the idea that NLEs can ameliorate disease conditions involving chronic inflammation.

Chemical Constituents of Nelumbo nucifera Seeds

The phytochemical study for the extract of Nelumbo nucifera (Nymphaceae) seeds has led to the isolation of ten compounds including five simple phenolic compounds, two indole derivatives, a flavonoid glycoside, two abscisic acid derivatives. The interpretation of 1D and 2D NMR and ESI-Q-TOF-MS spectroscopic data revealed the chemical structures of isolates to be p-hydroxybenzoic acid (1), protocatechuic acid (2), (E)-p-coumaric acid (3), (E)-ferulic acid (4), (E)-sinapate-4-O-β-D-glucopyranoside (5), tryptophan (6), 3-indoleacetic acid (7), isoschaftoside (8), dihydrophaseic acid (9), dihydrophaseic acid 3′-O-β-D-glucopyranoside (10). To the best of our knowledge, 1 – 5 and 7 were identified for the first time from N. nucifera seeds, and the presence of dihydrophaseic acid (9) and its glucoside (10) were demonstrated secondly in this plant.

Study on chemical constituents in stems of Nelumbo nucifera by UPLC-ESI/Q-TOF-MS/MS / 中国中药杂志

This paper employed UPLC-Electrospray Ionization /Quadrupole-Time of Flight-Mass /Mass Spectrometry( UPLC-ESI/Q-TOF-MS/MS) to analyze the chemical constituents in the stems of Nelumbo nucifera. The stems of N. nucifera were extracted with 75% methanol, and we applied an Agilent Zorbax SB-Aq column (2.1 mm x 100 mm, 1.8 μm) to UPLC analysis with water methanol-water( containing 0.05% formic acid) in gradient as mobile phase. The eluates were then detected by ESI-Q-TOF-MS/MS. Results indicated that 22 benzylisoquinoline alkaloids were indendified. Among them, one alkaloid may be a new compound and a component was found in the Lotus for the first time. We fully identify the composition of the Lotus stems for the first time, Which could provides theoretical foundation for further study and utilization of the medicinal resources.

Study on content determination of alkaloids and HPLC fingerprint of "Jianlian" Nelumbinis Plumula / 中国中药杂志

This study is to determine the content of three alkaloids and establish the HPLC fingerprint of "Jianlian" Nelumbinis Plumula. The HPLC method of content determination was as follows: Thermo C18 (4. 6 mm x 250 mm, 5 μm) was conducted with acetonitrile-sodium dodecyl sulfonate solution-acetic acid (56: 43: 1) at a flow rate of 1.0 mL x min(-1). The monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. The method of HPLC fingerprint was as follows: Agilent ZORBAX SB-Aq C18 (4.6 mm x 250 mm, 5 μm) was conducted with gradient elution of methanol and water at a flow rate of 0.8 mL x min(-1), the monitoring wavelength was set at 282 nm and the column temperature was 35 degrees C. Similarities evaluation and hierarchical clustering analysis were applied to demonstrate the variability of 12 batches of "Jianlian" Nelumbinis Plumula samples. The results demonstrated that 11 batches showed good similarity on chemical constituents. The method could well display the chemical information of "Jianlian" Nelumbinis Plumula. It was simple, reliable and could be used for the chemical quality control of "Jianlian" Nelumbinis Plumula.

Studies on flavonoids from stems of Nelumbo nucifera Gaertn and their cytotoxic activities / 中国中药杂志

This research is to investigate study the flavonoids from stems of Nelumbo nucifera and the cytotoxic activities of iso- lated compounds. The constituents were separated by column chromatography,and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for cytoxic activities by MTT method. Twelve compounds were isolated and identified as rhamnazin-3-O-beta-D-glucopyranoside (1), luteolin-3', 4'-dimethylether-7-O-beta-D-glucoside (2), kaempferol-3-O-beta-D-xylopyranosyl-(1-->2)-O-beta-D-glucopyranoside (3), quercetin-3,3'-di-O-beta-D-glucopyranoside (4), 1, 8-dihydroxy-3,7-dimethoxyxanthone (5), isorhamnetin-3-O-beta-D-glucopyranoside(6) , kaempferol(7), isorhamnetin (8), quercetin(9), astragalin(10), hyperoside (11) and 1-hy- droxy-3,7,8-trimethoxyxanthone(12). All compounds were isolated from stems of this plant for the first time, and compounds 1-5 were firstly isolated from the family nelumbonaceae. Compounds 24 and 6 showed significant cytotoxic activities against BEL-7402 carcinoma cell lines at a concentration of 1 x 10(-5) mol x L(-1) with the inhibitory rate of 67.36%, 53.25%, 57.78%, 60.13% and 52.11%, respectively.

Chemical constituents from leaves of Nelumbo nucifera / 中国中药杂志

To study the chemical constituents, twenty-seven compounds were isolated from the 70% ethanol extract from leaves of Nelumbo nucifera by modern chromatographic techniques. Their structures were identified as 10-octacosanol (1), beta-sitosterol (2), 1-undecanol (3), 1-eicosanol (4), daucosterol (5), 6'-hydroxy-4,4'-dimethoxychalcone (6), 3,7,8-trimethoxy-1-hydroxy-xanthone (7), rhamnetin-3-O-beta-D-glucopyranoside (8), chrysoeriol-7-O-beta-D-glucoside (9), quercetin-3-O-beta-D-glucopyranoside (10), quercetin-3-O-alpha-L-rhamnopyranosyl (11), hyperoside (12), quercetin-3-O-rutinoside (13), astragalin (14), isorhamnetin-3-O-alpha-L-rhamnopyranosyl-(1--> 6)-[alpha-D-lyxopyranosyl-(1 --> 2) -beta-D-glucopyranoside] (15), isorhamnetin-3-O-alpha-D-lyxopyranosyl-(1 --> 2) -beta-D-glucopyranoside (16), isorhamnetin-3-O-beta-D-glucopyranoside (17), isorhamnetin-3-O-alpha-L-rhamnopyranosyl-(1 --> 6)-beta-D-glucopyranoside (18), quercetin (19), kaempferol (20), dehydronuciferine (21), roemerine (22), stigmast-7-en-3-O-beta-D-glucopyranoside (23), stigmast-7-en-3beta-ol (24), and benzene-1,2-diol (25) on the basis of spectral data analysis. Compounds 1, 6, 7, 8, 24 and 25 were isolated from this plant for the first time, and compounds 15-18 were isolated from the leaves for the first time. Compounds 6, 8, 10, 11, 13 and 15 showed inhibitory activities against beta amyloid (1-42) by A-beta aggregation method with inhibition rates of (63.99 +/- 24.29)%, (79.61 +/- 4.49)%, (49.96 +/- 12.61)%, (101.19 +/- 8.19)%, (88.41+/-6.76)% and (72.48 +/- 8.97)%, respectively.

Cytotoxic alkaloids from stems of Nelumbo nucifera / 中国中药杂志

Chemical investigation was carried out to study the alkaloids from stems of Nelumbo nucifera and their cytotoxic activities. The constituents were separated by column chromatography, and their structures were elucidated by spectroscopic data analyses. The isolated compounds were evaluated for their cytotoxic activities by MTr method. Fifteen compounds were isolated from the total alkaloids extract and identified as asimilobine (1), isococlaurine (2), N-acetylnorarmepavine (3), crykonisine (4), velucryptine (5), pycnarrhine (6), liriodenine (7), nuciferine (8), nornuciferine (9), armepavine (10), N-methylasimilobine (11), coclaurine (12), N-norarmepavine (13), N-methylcoclaurine (14) and lysicamine (15). Compounds 1-7 and 12-15 were isolated from stems of this plant for the first time, and compounds 2-6 were firstly isolated from the family Nelumbonaceae. Compounds 7-10, 13 and 14 showed significant cytotoxic activities against HL-60 carcinoma cell line with inhibitory ratios of 51.36%, 59.09%, 52.51%, 53.93%, 51.43%, and 64.31% at concentration of 1 x 10(-5) mol x L(-1), respectively.

Screening of pancreatic lipase and alpha-glucosidase inhibitors from Chinese dietary herbs / 中国中药杂志

The present study was conducted to develop new inhibitors of pancreatic lipase and alpha-glucosidase from Chinese dietary herbs. Sixty-three dietary herbs from 39 taxonomic families were selected and extracted with aqueous ethanol or water. The extracts were then tested with in vitro enzyme assays for their ability to inhibit pancreatic lipase and alpha-glucosidase activities. Orlistat and acarbose were used as two positive controls. The extracts of Nelumbo nucifera, Curcuma longa, Piper longum and Morus alba showed strong pancreatic lipase inhibitory effects with IC50 at (28.00 +/- 5.51), (5.24 +/- 0.51), (14.76 +/- 2.58), (4.78 +/- 0.58), (3.41 +/- 0.67) mg x L(-1), respectively. These extracts also showed potent alpha-glucosidase inhibitory activities with IC50 at (1.98 +/- 0.13), (0. 18 + 0.007), (0.71 +/- 0.08), (0.077 +/- 0.005), (0.089 +/- 0.006) g x L(-1), respectively. The results provide useful information for developing new drugs or natural health products for hyperlipidemia and hypoglycemia from Chinese dietary herbs.

Effects of plant extract neferine on cyclic adenosine monophosphate and cyclic guanosine monophosphate levels in rabbit corpus cavernosum in vitro / 亚洲男科学杂志(英文版)

<p><b>AIM</b>To further investigate the relaxation mechanism of neferine (Nef), a bis-benzylisoquinoline alkaloid extracted (isolated) from the green seed embryo of Nelumbo nucifera Gaertn in China, on rabbit corpus cavernosum tissue in vitro.</p><p><b>METHODS</b>The effects of Nef on the concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) in isolated and incubated rabbit corpus cavernosum tissue were recorded using 125I radioimmunoassay.</p><p><b>RESULTS</b>The basal concentration of cAMP in corpus cavernosum tissue was 5.67 +/- 0.97 pmol/mg. Nef increased the cAMP concentration in a dose-dependent manner (P < 0.05), but this effect was not inhibited by an adenylate cyclase inhibitor (cis-N-[2-phenylcyclopentyl]azacyclotridec-1-en-2-amine, MDL-12, 330A) (P > 0.05). The accumulation of cAMP induced by prostaglandin E1 (PGE1, a stimulator of cAMP production) was also augmented by Nef in a dose-dependent manner (P < 0.05). The basal concentration of cGMP in corpus cavernosum tissue is 0.44 +/- 0.09 pmol/mg. Nef did not affect this concentration of cGMP, either in the presence or in the absence of a guanyl cyclase inhibitor (1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, ODQ) (P > 0.05). Also, sodium nitroprusside (SNP, a stimulator of cGMP production)-induced cGMP production was not enhanced by Nef (P > 0.05).</p><p><b>CONCLUSION</b>Nef, with its relaxation mechanism, can enhance the concentration of cAMP in rabbit corpus cavernosum tissue, probably by inhibiting phosphodiesterase activity.</p>

HPLC determination of 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine in Nelumbo nucifera and its alkaloid fraction / 中国中药杂志

<p><b>OBJECTIVE</b>To establish an HPLC method for the determination of four alkaloids, i.e., 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine, in Nelumbo nucifera and its alkaloid fraction.</p><p><b>METHOD</b>The determination was carried out at 35 degrees C on a Hypersil C18 column (4.6 mm x 250 mm, 5 microm), eluting with acetonitrile-water containing 0.1% triethylamine as mobile phases in gradient mode. The flow rate was 1.0 mL x min(-1) and detection at the wavelength was set at 270 nm.</p><p><b>RESULT</b>The linear ranges of 2-hydroxy-1-methoxyaporphine, pronuciferine, nuciferine and roemerine were 0.110-0.658 microg (r = 0.9995), 0.0210-0.126 microg (r = 0.9995), 0.103-0.618 microg (r = 0.9998), 0.085 6-0.514 microg (r = 0.9995), with the average recoveries (n=6) were 101.5%, 99.14%, 99.21% and 98.41% for the alkaloid fraction of N. nucifera and 99.53%, 100.5%, 97.51% and 100.1% for N. nucifera respectively.</p><p><b>CONCLUSION</b>The determination results of the three batches of samples showed that the method was easy and accurate which could be used to determine the contents of four components in N. nucifera and its alkaloid fraction.</p>

Study on preparation technology of plumula nelumbinis Alkaloid Dripping Pill and determination of content by HPLC / 中国中药杂志

<p><b>OBJECTIVE</b>To determine the optimum technical parameters through controlling the different factors and the content of Dripping Pill.</p><p><b>METHOD</b>Time of dissolution, spherical degree and pill weight as parameters, the influential factors were investigated by orthogonal test and then to determine the content of liensinine, isoliensinine, neferine in plumula nelumbinis Alkaloid Dripping Pill.</p><p><b>RESULT</b>The optimum condition: 75% PEG4000 as matrix, methyl-silicon oil as refrigerant, 4 cm dripping distance, 10 degrees C refrigerant, 30 drippings per minute.</p><p><b>CONCLUSION</b>The good quality pills can be produced through this process and the HPLC determination method is simple, reliable and can be used in the quality control of plumula nelumbinis Alkaloid Dripping Pill.</p>

Molecular identity of Crocus sativus and its misused substitutes by ITS sequence / 中国中药杂志

<p><b>OBJECTIVE</b>To find the patterns of the rDNA ITS sequence variation of Crocus sativus, Chrysanthemum chanetii, Nelumbo nucifera, Zea mays and Garthamus tinctorius and to establish the molecular biological method for the identification of C. sativus and the others.</p><p><b>METHOD</b>After the total DNA of Crocus sativus, C. vernus-w and C. vernus-p were extracted, the ITS sequence was amplified by PCR with universal primer of ITS and PCR product was sequenced after purification and cloning. The ITS sequences of Chrysanthemrnum chanetii, Nelumbo nucifera, Zea mays and Garthamus tinctorius were obtained from GenBank.</p><p><b>RESULT</b>The complete ITS sequence of Crocus sativus, C. vernus-w and C. vernus-p, including ITSI rDNA, 5.8S rDNA, ITS2 rDNA were measured. The GenBank accession No. was DQ094185, DQ224363 and DQ224364 respectively. The similarity of ITS sequence between C. sativus and the two garden species of C. vernus was above 91%; the identity was 99.84% between C. vernus-w and C. vernus-p. The range of diversity between C. sativus and other herbs was above 46% based on ITS1 and above 41% based on ITS2.</p><p><b>CONCLUSION</b>C. sativus can be distinguished from misused substitutes by the ITS sequence. The ITS sequence is an available molecular marker for identification of the C. sativus.</p>

PCR, clone and sequence analysis of rDNA-ITS of Nelumbo nucifera from different geographical origins in China / 中国中药杂志

<p><b>OBJECTIVE</b>To provide DNA molecular marker for identification of Nelumbo nucifera by exploring the differences of nrDNA-ITS sequence of N. nucifera originated from different habitats.</p><p><b>METHOD</b>To compare nrDNA-ITS base sequence using specific PCR-ITS.</p><p><b>RESULT</b>The completed sequence of ITS and 5.8 S rDNA, and the partial sequences of 18S rDNA and 26S rDNA, totally 750 bp, from N. nucifera were obtained. The differences among N. nucifera from different habitats and from different cultivars were found.</p><p><b>CONCLUSION</b>The method can be used to identify N. nucifera among different species and to distinguish their fakes. It provided the basis for identifying N. nucifera from different geographical regions by comparison of their ITS sequences.</p>

Metabolic products and pathway of neferine in rat liver / 药学学报

The present study utilized LC-MS and HPLC approaches to characterize the metabolites of neferine in rat liver after an oral administration of 20 mg x kg(-1), and investigated the involvement of CYP450 isoforms in the metabolism of neferine by their selective inhibitors in vitro, separately. In positive ionization mode, besides neferine, four metabolites (M1-M4) were detected. M2 (the major metabolite) and M4 were identified as liensinine and isoliensinine by comparison with reference substances. Moreover, according to the analysis of metabolic rule of parent drug (neferine), M1 and M3 may be desmethylliensinine and desmethyl-isoliensinine, respectively. Furthermore, the metabolism of neferine in rat liver microsomes showed that the percentage inhibition of the major metabolism (liensinine) formation was 80.5% by quinidine (10 micromol x L(-1), selective CYP2D1 inhibitor) and 25.7% by ketoconazole (1 micromol x L(-1), selective CYP3A1 inhibitor). Neferine was mainly metabolized by CYP2D1 or CYP3A1 to liensinine, isoliensinine, desmethyl-liensinine and desmethyl-isoliensinine.

Separation and purification processes of alkaloids from Lotus plumule with macroporous adsorption resin / 中国中药杂志

<p><b>OBJECTIVE</b>To explore the optimal separation of the alkaloids from Lotus plumule by selecting appropriate macroporous adsorption resins.</p><p><b>METHOD</b>To evaluated the separating efficiency by measuring the adsorption ratio, eluting ratio and the concentration of neferine.</p><p><b>RESULT</b>The LSA -5B macroporous adsorption resin prolided the optimal separating efficiency. The best adsorption capacity was achieved with the following conditions: the concentration of extact liquid was 0. 125 g x mL(-1) (equivalence to raw material), then the resin was washed by water to remove impurity and alkaloids were desorbed by 50% ethanol.</p><p><b>CONCLUSION</b>This method is simple, feasible and suitable for industry production.</p>

Antimicrobial potential of certain medicinal plants available in Western Ghats of India

Ethanol extracts from the plant Hypericum mysorense [Family: Hypericaceae]. Passiflora edulis [Family: Passifloraceae] and Nelumbo nucifcra Gaertn. [Family: Nymphaeaceae] were screened for antimicrobial activity. All the extracts showed that almost all concentration [40 mg/ml, 20 mg/ml, 10 mg/ml] of extracts possesses antibacterial and antifungal activity against three gram +ve, three gram -ve and three fungal strains by zone of inhibition method

Effects of isoliensinine on proliferation of porcine coronary arterial smooth muscle cells induced by phenylephrine / 药学学报

<p><b>AIM</b>To investigate the inhibitory effects and mechanism of action of isoliensinine (IL) on the proliferation of porcine coronary arterial smooth muscle cells (CASMCs) induced by phenylephrine (Phen) and its mechanisms of action.</p><p><b>METHODS</b>MTT assay, immunohistochemical method and Western blotting were adopted.</p><p><b>RESULTS</b>IL (0.03 - 3 micromol x L(-1)) could inhibit the CASMCs proliferation induced by Phen (0.1 micromol x L(-1)) in a concentration-dependent manner. IL (0.1 micromol x L(-1)) antagonized Phen-induced overexpression of PDGF-beta and bFGF from 0.545 +/- 0.026 and 0.47 +/- 0.03 to 0.458 +/- 0.019 and 0.376 +/- 0.017 (P < 0.01 , P < 0.01). IL (0.1 micromol x L(-1)) also decreased c-fos, c-myc and hsp70 overexpression induced by Phen from 0.57 +/- 0.04, 0.44 +/- 0.04 and (173 +/- 36)% to 0.46 +/- 0.05, 0.372 +/- 0.021 and (115 +/- 35)% respectively (P < 0.01, P < 0.01, P < 0.01).</p><p><b>CONCLUSION</b>IL exerted antiproliferative effect on CASMCs induced by phenylephrine, and its mechanisms were related to decrease the overexpression of growth factors (PDGF-beta, bFGF), protooncogene (c-fos, c-myc) and hsp70.</p>