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Table of Contents
July-September 2014
Volume 10 | Issue 39
Page Nos. 191-383
Online since Friday, July 25, 2014
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ORIGINAL ARTICLES
Direct proof by
13
C-nuclear magnetic resonance of semi-purified extract and isolation of
ent
-Catechin from leaves of
Eucalyptus cinerea
p. 191
Sayonara Mendes Silva, Simone Yae Abe, Fernanda Giacomini Bueno, Norberto Peporine Lopes, Joćo Carlos Palazzo de Mello, Tomoe Nakashima
DOI
:10.4103/0973-1296.137355
PMID
:25210302
Background:
Eucalyptus cinerea
F. Muell. ex Benth. is native to Australia and acclimatized to Southern Brazil. Its aromatic leaves are used for ornamental purposes and have great potential for essential oil production, although reports of its use in folk medicine are few.
Objective:
This study evaluated the composition of
E. cinerea
leaves using the solid state
13
C-nuclear magnetic resonance (NMR) and isolation of the compound from the semipurified extract (SE).
Materials
and
Methods:
The SE of
E. cinerea
leaves was evaluated in the solid state by
13
C-NMR spectrum, and the SE was chromatographed on a Sephadex LH-20 column, followed by high-speed counter-current chromatography to isolate the compound. The SE was analyzed by
13
C-NMR and matrix-assisted laser desorption/ionization-time-of-flight spectra.
Results:
Flavan-3-ol units were present, suggesting the presence of proanthocyanidins as well as a gallic acid unit. The uncommon
ent
-catechin was isolated.
Conclusion:
The presence of
ent
-catechin is reported for the first time in this genus and species.
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Simultaneous determination of four neuroprotective compounds of
Tilia amurensis
by high performance liquid chromatography coupled with diode array detector
p. 195
Bohyung Lee, Jin Bae Weon, Bo-Ra Yun, Jiwoo Lee, Min Rye Eom, Choong Je Ma
DOI
:10.4103/0973-1296.137353
PMID
:25210303
Background:
Tilia amurensis
consists of various compounds, such as flavonoids and terpenoids.
Objective:
A simple and reliable high performance liquid chromatography (HPLC) coupled with the diode array detector (DAD) method has been established for simultaneous determination of epicatechin, nudiposide, lyoniside, and scopoletin isolated from
Tilia amurensis
.
Materials and Methods:
Optimum separations were obtained with a SHISEIDO C
18
column by gradient elution, with 0.1% Trifluoroacetic acid (TFA) water-methanol as the mobile phase. The gradient elution system was completed within 40 minutes. The flow rate and detection wavelength were 1 mL/minute, 205 nm, 250 nm, and 280 nm, respectively.
Results:
Validation of the analytical method was evaluated by linearity, precision, and the accuracy test. The calibration curve was linear over the established range with R
2
> 0.997. The limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.01-15.20 μg/mL and 0.03-46.06 μg/mL. The method exhibited an intraday and interday precision range of 96.25-105.66% and 93.52-109.92%, respectively (RSD <2.80%). The recoveries and relative standard deviation (RSD) of the four compounds in
Tilia amurensis
were in the range of 90.42-104.84% and 0.2-2.58%.
Conclusion:
This developed method was accurate and reliable for the quality evaluation of the four compounds isolated from
Tilia amurensis
.
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Anticancer activity of
cissampelos pareira
against dalton's lymphoma ascites bearing mice
p. 200
B Samuel Thavamani, Molly Mathew, SP Dhanabal
DOI
:10.4103/0973-1296.137356
PMID
:25210304
Background:
Cissampelos pareira
(Menispermaceae) is used in folk Indian system of alternative medicine, for its analgesic, antipyretic, diuretic, antilithic, and emmenagogue properties.
Objective:
To evaluate
Cissampelos pareira
(
C. pareira)
for
in vitro
cytotoxicity and
in vivo
antitumor activity against Dalton's Lymphoma Ascites (DLA) cells in Swiss mice.
Materials and Methods:
Cissampelos pareira
was successively extracted using different solvents.
In vitro
cytotoxicity was assessed by the MTT assay. An
in vivo
study was carried out in methanol extract. Twenty-four hours after intraperitoneal inoculation of the DLA cells in mice, the methanol extract of
C. pariera
(MECP) was administered at 200 and 400 mg/kg body weight for 14 consecutive days. On day 14, six mice were sacrificed and the rest were kept alive for assessment of increase in life-span. The antitumor effect was assessed by evaluating the packed cell volume, viable tumor cell count, increase in body weight, and increase in life-span. The hematological and serum biochemical parameters and anti-oxidant properties were assessed by estimating the superoxide dismutase (SOD), catalase (CAT), and lipid peroxidation.
Results:
Methanol Extract of
Cissampelos pariera
(MECP) showed a potent cytotoxic activity, with an IC
50
value of 95.5 μg/ml and a significant (
P
< 0.001) decrease in packed cell volume, viable cell count, and an increased lifespan (54 and 72%). The hematological and serum biochemical profiles were restored to normal levels in MECP-treated mice. The MECP-treated group significantly (
P
< 0.001) decreased SOD, lipid peroxidation, and CAT to normal.
Conclusion:
This study demonstrated that
C. pariera
exhibited significant
in vitro
and
in vivo
anti-tumor activities and that it was reasonably imputable to its increasing endogenous mechanism of antioxidant property.
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Comparison of toad skins
Bufo bufo gargarizans
Cantor from different regions for their active constituents content and cytotoxic activity on lung carcinoma cell lines
p. 207
Congyan Liu, Wei Cao, Yan Chen, Ding Qu, Jing Zhou
DOI
:10.4103/0973-1296.137358
PMID
:25210305
Background:
The skin of
Bufo bufo gargarizans
Cantor, rich in bufadienolides, peptides, and alkaloids, has approved pharmacological activity for preliminary anti-liver and lung tumor treatment. However, few studies have systematically focused on the influence of the producing regions on the content and antitumor activity of the active constituents in toad skins.
Objective:
This study aims to compare toad skins obtained from six different regions in China (Jiangsu, Anhui, Henan, Hebei, Jiangxi, and Shandong province) for their bufadienolide and alkaloid content, and their cytotoxic activity on two lung carcinoma cell lines (SPC-A-1 cells and A549 cells).
Materials and Methods:
High performance liquid chromatography (HPLC) was used to quantificationally determine four bufadienolides, which included bufotalin, bufalin, cinobufagin, and resibufogenin in toad skins, from six different regions, respectively. In addition, an ultraviolet (UV) spectrophotometer was also employed to identify the content of the total alkaloids using 5-hydroxytryptamine (5-HT) as the reference substance. An MTT assay was performed to compare the antiproliferative effects of the toad skins' ethanolic extracts from the different regions against SPC-A-1 and A549 cells.
Results:
In this study, the toad skins from Jiangsu province had the highest amount of bufadienolides (472.6 μg/g crude drug) and alkaloids (1.51 mg/g crude drug). Meanwhile, according to the extract, it exhibited the strongest cytotoxic effect against the lung carcinoma cell line (SPC-A-1 cells and A549 cells) with IC
50
values of 24.82 ± 0.76 and 23.77 ± 0.63 μg crude drug/mL, respectively.
Conclusion:
The toad skins that originated from the Jiangsu province, have comparatively greater advantages over samples from other regions as far as active constituent content and potential anti-lung cancer activity is concerned, suggesting that it can be a promising chemotherapeutic agent in lung cancer therapy, in further studies.
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New Sesquiterpene and Polymethoxy-Flavonoids from
Artemisia annua
L
p. 213
Yang Chu, Hongbo Wang, Jing Chen, Yue Hou
DOI
:10.4103/0973-1296.137359
PMID
:25210306
Our previous study revealed that the polymethoxy-flavonoids, as main components of
Artemisia annua,
could improve the antimalarial activity of
Artemisinin
. Here, we described the isolation, elucidation, constituent analysis, flavonoids enrichment of the extracts of
A
.
annua
. A total of 20 compounds were isolated including a new sesquiterpene (compound 12) and five (1, 5, 6, 7, 15) afforded for the first time from A. annua. The elucidation of eight flavonoids may be a useful phytochemical data and chemical foundation for further mechanism studies on improving the anti-malarial action of
artemisinin
. Furthermore, the antitumor activities of the compounds were assayed using four different kinds of human cancer cell lines.
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Preparation and pharmacokinetics in beagle dogs of ganershu sustained-release pellets
p. 217
Jin-huo Pan, Jian-chun Wang, Zhi-tao Jiang, Ting Zhang, Shao-bo Ge, Ye-xia Zhang, Xin Jin, Guo-jun Yan
DOI
:10.4103/0973-1296.137360
PMID
:25210307
Background:
The active ingredients of Ganershu compound recipe, which are effective for hepatitis treatment in liver protection and transaminase reduction. However, the active ingredients of Ganershu compound recipe are poor absorption, which conduct it has a low oral bioavailability.
Objective:
We prepared Ganershu sustained-release pellets (GSPs) by fluidized-bed on central composite design-response surface methodology and increase its bioavailability in beagle dogs.
Materials and Methods:
In this study, GSPs were successfully prepared. The Drug-loaded pellets and sustained-release coated were carried out in fluidized-bed machine. GSP was optimized for fitting release, roundness, and the overall desirability by central composite design-response surface methodology.
Results:
To optimize cumulative release profile, the outermost ethyl cellulose coating layer and the hydroxypropyl methyl cellulose (HPMC) swelling layer were employed, which were respectively given coating levels in terms of weight gain of 22% and 6%, the concentration of HPMC is 4.5% (g/ml). The pharmacokinetics of Ganershu normal pellets (GNPs) and GSP was studied in beagle dogs after oral administration. The naringenin as an index, the area under the curve
0-∞
of naringenin in GSP was 1.38 times greater than that of GNP. Meanwhile,
T
max of GSP was prolonged for about 74%.
Conclusion:
This study can clearly indicate that we enhanced the oral bioavailability of Ganershu by preparing the GSP, which had the sustained dissolution and improved the potential of it for clinical application.
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Stability indicating studies on NMITLI 118RT+ (standardized extract of
withania somnifera
dunal)
p. 227
Hafsa Ahmad, Kiran Khandelwal, Shakti Deep Pachauri, Rajender Singh Sanghwan, Anil Kumar Dwivedi
DOI
:10.4103/0973-1296.137361
PMID
:25210308
Background:
Withania somnifera Dunal
(Ashwagandha
) is an Indian medicinal plant of great medicinal value; used in many clinically proven conditions. NMITLI-118RT+ is a candidate drug under a Council of Scientific and Industrial Research (CSIR) networking project. It is a chemotype of
W. somnifera
's root extract, which has been used for the present study.
Objectives:
The present investigation aims to develop and validate a simple isocratic reverse phase-high performance liquid chromatography (RP-HPLC) system for the detection and estimation of Withanolide A (marker compound) and its analytical application for stability indicating studies on NMITLI-118RT+.
Material and Methods:
A validated RP-HPLC method for Withanolide A was established on a Waters HPLC system and the same was used on NMITLI-118RT+ for quantification and fingerprinting purposes, and for establishing forced degradation, isothermal stress tests, and drug-excipient testing protocols as per International Conference on Harmonization (ICH) guidelines.
Results:
A validated method was established, which could detect the marker at a retention time of around 6.3 minutes, with a linearity range of 2-100 μg/mL, by varying the amounts of the said marker, which were estimated in four different batches of NMITLI-118RT+. Photostability as per ICH guidelines suggested a slight loss of the active constituent and maximum degradation was afforded with alkali followed by acid, and then peroxide, in the forced degradation studies. In the drug-excipient studies, the maximum amount of active constituent could be detected in the samples with ethyl cellulose and the least with hydroxy propyl cellulose.
Conclusion:
The method developed here was simple and rapid. The various stability indicating studies carried out in the present investigation would be useful for formulation development and were suggestive of deciding the recommended storage conditions for NMITLI-118RT+.
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Efficient production of flavonoids in
Fagopyrum tataricum
hairy root cultures with yeast polysaccharide elicitation and medium renewal process
p. 234
Jiang-Lin Zhao, Liang Zou, Cai-Qiong Zhang, Yuan-Yuan Li, Lian-Xin Peng, Da-Bing Xiang, Gang Zhao
DOI
:10.4103/0973-1296.137362
PMID
:25210309
Background:
Tartary buckwheat (
Fagopyrum tataricum)
, an excellent edible and medicinal crop, has been widely used as a daily diet and traditional medicine for a long time. The major functional components of
Fagopyrum tataricum
have been demonstrated to be flavonoids (i.e. rutin and quercetin), which had notable andioxidant, antidiabetic, hypocholesterolemic and antitumor activities. Hairy root culture is a convenient and efficient plant tissue culture system for large scale production of bioactive metabolites.
Objective:
To enhance the functional flavonoids production in hairy root culture of
F. tataricum
.
Materials and Methods:
The elicitation treatment in combination with medium renewal strategy was applied for efficient promoting flavonoids production in
F. tataricum h
airy root cultures.
Results:
The exogenous yeast polysaccharide (YPS) elicitor notably stimulated the functional metabolites production in
F. tataricum
hairy root cultures, and the stimulation effect was concentration-dependent. Combination with the YPS elicitation (200 mg/L) and medium renewal process, the maximal flavonoids yield was enhanced to 47.13 mg/L, about 3.2-fold in comparison with the control culture of 14.88 mg/L. Moreover, this research also revealed the accumulation of these bioactive metabolites resulted from the stimulation of the phenylpropanoid pathway by YPS treatment. These results indicated that the
F. tataricum
hairy root culture could be an effective system for rutin and quercetin production.
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LC-MS/MS determination and pharmacokinetics study of puerarin and daidzein in rat plasma after oral administration of Gegenqinlian decoction and
Radix Puerariae
extract
p. 241
Yifan Zhang, Jin Yuan, Ying Wang, Yue Wang, Rui An, Xinhong Wang
DOI
:10.4103/0973-1296.137363
PMID
:25210310
Background:
Gegenqinlian decoction (GQD) is a famous traditional medicine recipe. It is composed of four herbs including
Radix Puerariae
(GG),
Radix Scutellariae
(HQ),
Rhizoma Coptidis
(HL) and
Radix Glycyrrhizae
(GC), which is widely used for treating gastro-intestinal disorders in the clinical practice of Traditional Chinese medicine (TCM). The aim of this study was to compare the pharmacokinetics of puerarin and daidzein in rats following oral administration of Gegenqinlian Decoction and
Radix puerariae
extract. Thus, a sensitive and selective liquid chromatography/tandem mass spectrometry (LC-MS/MS) method was developed and validated for simultaneous determination of puerarin and daidzein in rat plasma following oral administration of Gegenqinlian Decoction and
Radix Puerariae
extract.
Materials and Methods:
Chromatographic separation was performed on a Shiseido CAPCELL PAK C
18
analytical column (100 mm × 2.0 mm i.d., 5 μm) by linear gradient elution, with water (0.1% formic acid)-acetonitrile (0.1% formic acid) as mobile phase. Detection was carried out by multiple reaction monitoring (MRM) mode using electrospray ionization in the positive ion mode.
Results:
The calibration curves were linear over a range of 7.80-1560 ng/mL for puerarin and 6.30-1260 ng/mL for daidzein. The intra- and inter-day precision values were less than 13.6% and their average recoveries was in the range of 77.8% and 88.6% for puerarin and was between 76.3 and 86.8% for daidzein, respectively.
Conclusion:
The validated method was applied to the comparative pharmacokinetic studies of puerarin and daidzein after oral administration of Gegenqinlian Decoction and
Radix Puerariae
extract. The pharmacokinetic parameters showed that puerarin and daidzein from Gegenqinlian Decoction were absorbed more effectively with slower elimination in rat plasma than that from
Radix Puerariae
extract. These results revealed that as far as the
Radix Puerariae
extract was concerned, it is very valuable to be used as a clinical directions of Gegenqinlian Decoction.
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Fast analysis of principal volatile compounds in crude and processed
Atractylodes macrocephala
by an automated static headspace gas chromatography-mass spectrometry
p. 249
Jida Zhang, Gang Cao, Yunhua Xia, Chengping Wen, Yongsheng Fan
DOI
:10.4103/0973-1296.137364
PMID
:25210311
Objective:
Atractylodes macrocephala
, a famous herbal medicine, is used extensively in the practice of Traditional Chinese Medicine (TCM). Processing procedure is a common approach that usually occurs before
A. macrocephala
is prescribed. This paper describes a sensitive and specific assay for the determination of principal volatile compounds in crude and processed
A. macrocephala
.
Materials and Methods:
The present study concentrated on the development of a static headspace gas chromatography-mass spectrometry (SHS-GC/MS) for separating and identifying of volatile compounds from crude and processed
A. macrocephala
samples.
Results:
The results showed that the volatile oil in crude and processed
A. macrocephala
was markedly quantitatively and qualitatively different. Processing resulted in the reduction of volatile oil contents and variation of chemical compositions in
A. macrocephala
.
Conclusion:
The proposed method proved that SHS-GC/MS is rapid and specific, and should also be useful for evaluating the quality of crude and processed medicinal herbs.
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Qualitative and quantitative analysis of the major constituents in traditional Chinese medicine Danmu injection using LC-ESI-MS
n
and LC-DAD
p. 254
Fen-Xia Zhu, Jing-Jing Wang, Xiu-Feng Li, E Sun, Xiao-Bin Jia
DOI
:10.4103/0973-1296.137365
PMID
:25210312
Background:
Danmu injection, a traditional Chinese medicine (TCM) preparation made from
Nauclea officinalis
, has been commonly used for the treatment of cold, fever, swelling of throat in China. However, the chemical constituents in Danmu injection have not been clarified yet.
Objective:
a HPLC/DAD/ESI-MS
n
method was developed for qualitative and quantitative analysis of the components in Danmu injection.
Materials and Methods:
The chromatographic separation was performed on a Welch Material XB-C18 (4.6mm × 250mm, 5μm) using gradient elution with acetonitrile (A) and water containing 0.1% formic acid (B) as mobile phase at a flow rate of 1.0 ml/min.
Results:
Twenty-five compounds, including phenolic acid and phenol glycoside, iridous glycoside and glycoalkaloid were identified or tentatively deduced on the base of their retention behaviors, UV absorption, MS and MS
n
data with those elucidated references or literature. In addition, eleven compounds were simultaneously determined by HPLC-DAD, which was validated and successfully applied for determination of major components in Danmu injection.
Conclusion:
The results suggested that the established qualitative and quantitative method would be a powerful and reliable analytical tool for the characterization of multi-constituent in complex chemical system and quality control of Danmu injection.
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Rapid and undamaged analysis of crude and processed
Radix Scrophulariae
by Fourier transform infrared spectroscopy coupled with soft independent modeling of class analogy
p. 265
Huiping Zhu, Gang Cao, Hao Cai, Baochang Cai, Jue Hu
DOI
:10.4103/0973-1296.137366
PMID
:25210313
Objective:
The main objective of this work is to determine the feasibility of identification of crude and processed
Radix
Scrophulariae
using the Fourier transform infrared spectroscopy couple with soft independent modeling of class analogy (FT-IR-SIMCA).
Materials
and
Methods:
A total of 50 different crude
Radix
Scrophulariae
was used to product processed ones. The spectra were acquired by FT-IR spectroscopy using a diffuse reflectance fiber optic probe. For the multivariate analysis, SIMCA was used. Results showed that FT-IR-SIMCA was useful to discriminate the processed
Radix
Scrophulariae
samples from crude samples. These samples could be successfully classified by SIMCA.
Results:
In all cases, the recognition and rejection rates were 97.8% and 100%, respectively. When testing with the blind sample that was picked out from the chosen samples, the accuracy was up to 90%.
Conclusion:
It means that the methodology is capable of accurately separating processed
Radix
Scrophulariae
from crude samples.
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Antidepressant-like effects of Chaihu-Shugan-San via SAPK/JNK signal transduction in rat models of depression
p. 271
Yun-Hui Li, Chun-Hu Zhang, Juan Qiu, Su-E Wang, Sui-Yu Hu, Xi Huang, Ying Xie, Yang Wang, Tian-Li Cheng
DOI
:10.4103/0973-1296.137367
PMID
:25210314
Background:
Chaihu-Shugan-San (CHSGS), a traditional Chinese medicinal herbal formula, registered in Jingyue Quanshu, has been indicated that oral administration of the extract from it can remit depressive disorder. C-Jun amino-terminal kinase (JNK/SAPK) signal transduction plays a key role in the apoptosis of nerve cells, be reported closely correlated with depression. This study was designed to investigate CHSGS antidepressant-like effects in rat models of depression and probe its possible mechanism.
Materials and Methods:
The classical experimental depression model chronic mild unpredictable stress (CMUS) was used to evaluate the antidepressant-like effects of CHSGS. The extracts were administered orally for 14 days, while the parallel positive control was given at the same time using fluoxetine hydrochloride. The expressions of JNK in the hippocampus were detected by real-time fluorescent quantitation PCR and Western blot assay.
Results:
Intragastric administration of CHSGS for 14 days caused a significant improvement of weight and locomotor activity in the open-field test. In addition, CHSGS treatment inhibited the expressions of JNK in the hippocampus tissue in CMUS rats.
Conclusion:
CHSGS could obviously improve the depressive state of the model rats and its mechanism may be correlated with regulating the expressions of JNK in the hippocampus.
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Content determination of the flavonoids in the different parts and different species of
Abelmoschus esculentus
L
.
by reversed phase-high performance liquid chromatograph and colorimetric method
p. 278
Yin Lin, Min-feng Lu, Hai-bing Liao, Yu-xian Li, Wei Han, Ke Yuan
DOI
:10.4103/0973-1296.137368
PMID
:25210315
Background:
This research will establish the ultraviolet colorimetric method to determine the total flavonoid content in different species and different parts of
Abelmoschus
esculentus
L.
Materials and Methods:
We establish the reversed-phase high-performance liquid chromatograph (RP-HPLC) method to determine the content of the three flavonoid glycosides in different species and different parts of the
A.
esculentus
. Adopt the NaNO
2
-Al (NO
3
)
3
-NaOH colorimetric method to determine the total flavonoid content; at the same time, adopt the RP-HPLC method to determine the contents of the three flavonoid glycosides. Using the methods of ultraviolet colorimetry and RP-HPLC, we determined and analyzed the total flavonoid content and the content of the three flavonoid glycosides in different species and different parts of
A.
esculentus
.
Results:
There are great distribution differences of the total flavonoids and the three flavonoid glycosides in different species and parts of
A.
esculentus
. Among them, the content of the effective constituents in the flower is relatively high, next is in the fruit. In the different species of
A.
esculentus
, the content of the flavonoids of finger relatively high. The HPLC method established in this research is simple and convenient and its results are accurate and reliable. In addition, it has a very good repeatability.
Conclusion:
The results provided the reference data for the medicinal use of
A.
esculentus
and it can be used in quality analyzing of its effective constituents.
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Pharmacokinetics and tissue distribution study of tanshinone IIA after oral administration of Bushen Huoxue Qubi granules to rats with blood stasis syndrome
p. 285
Yuanqing Wang, Jianye Yan, Shunxiang Li, Xiong Cai, Wei Wang, Kun Luo, Dan Huang, Jiesheng Gao
DOI
:10.4103/0973-1296.137369
PMID
:25210316
Background:
Bushen Huoxue Qubi (BHQ) granules, a traditional Chinese medicine preparation, has been clinically used for the treatment of the blood stasis syndrome.
Objective:
The main objective is to investigate whether the diseased condition would alter the pharmacokinetics and tissue distribution of tanshinone IIA in BHQ, which was given orally to the acute blood stasis rats.
Materials
and
Methods:
The main bioactive constituent in BHQ, tanshinone IIA, was measured in the plasma and tissues of animals by the high performance liquid chromatography with ultraviolet detection. The analysis was successfully performed on an Agilent TC-C
18
column (250 × 4.6 mm I.D., 5 μm) protected with a Octadecylsilane (ODS) guard column (10 × 4.6 mm I.D., 5 μm). The mobile phase was aqueous solution (A) (containing 0.40% aqueous acetic acid) and acetonitrile (B). The conditions of the solvent gradient elution were 35-40% (B) in 0-15 min, 40-42% (B) in 15-18 min and 42-70% (B) in 18-30 min at a flow rate of 1.0 mL/min. Detection was conducted with wavelength of 270 nm at 30°C.
Results:
Good linearity relationships were found (
r
2
> 0.9955) over the investigated concentration range for bio-samples. Blood stasis was associated with significantly higher area under the concentration-time curve (AUC), the maximum plasma concentration (C
max
) and biological half-life (t
1/2
), lower total body clearance (CL) and apparent volume of distribution (Vd) of tanshinone IIA in plasma and higher AUC
0-t
of tanshinone IIA in the analyzed tissues of rats treated with BHQ.
Conclusion:
Blood stasis could alter pharmacokinetics and tissue distribution of tanshinone IIA in BHQ.
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Optimization of simultaneous ultrasonic-assisted extraction of water-soluble and fat-soluble characteristic constituents from Forsythiae Fructus Using response surface methodology and high-performance liquid chromatography
p. 292
Yong-Gang Xia, Bing-You Yang, Jun Liang, Di Wang, Qi Yang, Hai-Xue Kuang
DOI
:10.4103/0973-1296.137370
PMID
:25210317
Background:
The compounds (+)-pinoresinol-
β
-glucoside (1) forsythiaside, (2) phillyrin (3) and phillygenin (4) were elucidated to be the characteristic constituents for quality control of Forsythiae Fructus extract by chromatographic fingerprint in 2010 edition of Chinese Pharmacopoeia due to their numerous important pharmacological actions. It is of great interest to extract these medicinally active constituents from Forsythiae Fructus simultaneously.
Materials and Methods:
In this study, a new ultrasound-assisted extraction (UAE) method was developed for the simultaneous extraction of biological components 1-4 in Forsythiae Fructus. The quantitative effects of extraction time, ratio of liquid to solid, extraction temperature, and methanol concentration on yield of these four important biological constituents from Forsythiae Fructus were investigated using response surface methodology with Box-Behnken design. The compounds 1-4 extracted by UAE were quantitative analysis by high-performance liquid chromatography-photodiode array detect (HPLC-PAD), and overall desirability (OD), the geometric mean of the contents of four major biological components, was used as a marker to evaluate the extraction efficiency.
Results:
By solving the regression equation and analyzing 3-D plots, the optimum condition was at extraction temperature 70°C, time 60 min, ratio of liquid to solid 20, and methanol concentration 76.6%. Under these conditions, extraction yields of compounds 1-4 were 2.92 mg/g, 52.10 mg/g, 0.90 mg/g and 0.57 mg/g, respectively, which were in good agreement with the predicted OD values. In order to achieve a similar yield as UAE, soxhlet extraction required at least 6 h and maceration extraction required much longer time of 24 h. Established UAE method has been successfully applied to sample preparation for the quality control of Forsythiae Fructus. Additionally, a quadrupole time-of-flight mass spectrometry was applied to the structural confirmation of analytes from the complex matrices acquired by UAE.
Conclusion:
The results indicated that UAE is an effective alternative method for extracting bioactive constituents, which may facilitate a deeper understanding of the extract of active constituents in Forsythiae Fructus from the raw material to its extract for providing the theoretical references.
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Analysis of the influence of sulfur-fumigation on the volatile components of
Angelicae sinensis
Radix by comprehensive two-dimensional gas chromatography/time-of-flight mass spectrometry
p. 304
Gang Cao, Hao Cai, Yajing Lou, Sicong Tu, Xiao Liu, Kunming Qin, Baochang Cai
DOI
:10.4103/0973-1296.137371
PMID
:25210318
Background:
Sulfur-fumigation of
Angelicae
sinensis
Radix causes changes in the structure and composition of volatile components. These changes alter the curative effect and the quality of
A.
sinensis
Radix.
Materials and Methods:
In this study, comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC × GC-TOFMS) was employed to investigate the influence of sulfur-fumigation on the volatile components, and to characterize and quantify the chemical composition of the volatile oil of
A.
sinensis
Radix.
Results:
The present study has shown that sulfur-fumigated
A.
sinensis
Radix samples had significant loss of the main active compounds and a more destructive fingerprint profile compared to non-fumigated samples.
Conclusion:
From this study, it can be concluded that the combination of GC × GC and TOFMS has potential as a quality monitoring tool in herbal medicine and food processing industries.
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Antitumor effects of dammarane-type saponins from steamed Notoginseng
p. 314
Fan He, Yan Ding, Chun Liang, Seok Bean Song, De-Qiang Dou, Gyu Yong Song, Young Ho Kim
DOI
:10.4103/0973-1296.137372
PMID
:25210319
Six dammarane-type saponins were extracted from steamed
Panax notoginseng
. Their chemical structures were identified spectroscopically as ginsenosides Rh
1
(1), Rg
1
(2), 20 (S)-Rg
3
(3), 20 (R)-Rg
3
(4), Rb
3
(5), and Rb
1
(6). Compounds (0.1-10 μM) were tested for inhibition of tumor necrosis factor-α (TNF)-induced nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) luciferase reporter activity using a human kidney 293T cell-based assay. Ginsenoside Rb
3
(5) showed the most significant activity with an IC
50
of 8.2 μM. This compound also inhibited the induction of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) messenger Ribonucleic acid (mRNA) in a dose-dependent manner after HepG2 cells had been treated with TNF-α (10 ng/mL).
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Methyl protodioscin induces G2/M cell cycle arrest and apoptosis in A549 human lung cancer cells
p. 318
Yang Bai, Xiao-Yuan Qu, Jun-Qiang Yin, Liangcai Wu, Hong Jiang, Han-Wu Long, Qiang Jia
DOI
:10.4103/0973-1296.137373
PMID
:25210320
Background:
Methyl protodioscin (MPD) is a furostanol bisglycoside with antitumor properties. It has been shown to reduce proliferation, cause cell cycle arrest.
Objective:
The present study elucidates the mechanism underlying MPD's apoptotic effects, using the A549 human lung cancer cell line.
Materials and Methods:
The human pulmonary adenocarcinoma cell line A549 was obtained from the Cell Bank of the Animal Experiment Center, North School Region, Sun Yat-Sen University. All of the cells were grown in RPMI 1640 supplemented with 10% fetal calf serum (Hyclone, Logan, UT, USA), penicillin (10,000 U/l), and streptomycin (100 mg/l) at 37°C in a 5% CO
2
humidified atmosphere. The induction of apoptosis was observed in flow cytometry and fluorescent staining experiments.
Results:
MPD showed growth inhibitory effects in A549 cells in a dose- and time-dependent manner. The significant G2/M cell cycle arrest and apoptotic effect were also seen in A549 cells treated with MPD. MPD-induced apoptosis was accompanied by a significant reduction of mitochondrial membrane potential, release of mitochondrial cytochrome c to cytosol, activation of caspase-3, downregulation of Bcl-2, p-Bad, and upregulation of Bax.
Conclusion:
Our results show that the induction of apoptosis by MPD involves multiple molecular pathways and strongly suggest that Bcl-2 family proteins signaling pathways. In addition, mitochondrial membrane potential, mitochondrial cytochrome c and caspase-3 were also closely associated with MPD-induced apoptotic process in human A549 cells.
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Induction of apoptosis by
Cordyceps militaris
fraction in human chronic myeloid leukemia K562 cells involved with mitochondrial dysfunction
p. 325
Tian Tian, Liyan Song, Qin Zheng, Xianjing Hu, Rongmin Yu
DOI
:10.4103/0973-1296.137374
PMID
:25210321
Background:
Cordyceps militaris
is widely used for various ethno medical conditions including cancer and inflammation complications in traditional Chinese medicine.
Objective:
To investigate the
in vitro
antitumor activity of
Cordyceps militaris
fraction (CMF) and the molecular mechanism underlying the apoptosis it induces in human chronic myeloid leukemia K562 cells.
Materials and Methods:
CMF was prepared according to our previous report. Cell viability was assessed by MTT assay. The rate of apoptosis, distribution of cell cycle and loss of mitochondrial membrane potential were measured by flow cytometry. Caspase activities were analyzed by Western blot and oxygen consumption rate was recorded using the Oxytherm system.
Results:
The results demonstrated that CMF triggered growth inhibition in K562 cells with only minor toxicity on a normal human cell line and inhibited the proliferation of K562 cells in a dose- and time-dependent manner with IC
50
value of 34.1 ± 2.0 μg/ml after 48 h incubation. This most likely resulted from cell cycle arrest at the S phase and the induction of apoptosis. In addition, CMF induced activation of caspase-3 and subsequent cleavage of poly ADP-ribose polymerase (PARP). The caspase signals may originate from mitochondrial dysfunction, which was supported by the finding of decreased mitochondria transmembrance potential and the lower oxygen consumption rate.
Conclusion:
CMF possessed the
in vitro
antitumor effect on K562 cells and CMF-induced apoptosis might be involved by the mitochondrial dysfunction and valuable to research and develop as a potential antitumor agency.
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Enhanced dissolution and stability of Tanshinone IIA base by solid dispersion system with nano-hydroxyapatite
p. 332
Yan-rong Jiang, Zhen-hai Zhang, Sai-yan Huang, Yan Lu, Tian-tian Ma, Xiao-bin Jia
DOI
:10.4103/0973-1296.137375
PMID
:25210322
Background:
Tanshinone IIA (TSIIA) exhibits a variety of cardiovascular effects; however, it has low solubility in water. The preparation of poorly soluble drugs for oral delivery is one of the greatest challenges in the field of formulation research. Among the approaches available, solid dispersion (SD) technique has proven to be one of the most commonly used these methods for improving dissolution and bioavailability of drugs, because of its relative simplicity and economy in terms of both preparation and evaluation.
Objective:
This study was aimed at investigating the dissolution behavior and physical stability of SDs of TSIIA by employing nano-hydroxyapatite (n-HAp).
Materials
and
Methods:
The TSIIA SDs was prepared to use a spray-drying method. First, an
in vitro
dissolution test was performed to assess dissolution characteristics. Next, a set of complementary techniques (differential scanning calorimetry, scanning electron microscopy, X-ray powder diffraction, and Fourier transform infrared spectroscopy) was used to monitor the physicochemical properties of the SDs. The SDs was stored at 40°C/75% relative humidity for 6 months, after which their stability was assessed.
Results:
TSIIA dissolution remarkably improved because of the formulation of the SDs with n-HAp particles. Comparisons with the corresponding physical mixtures revealed changes in the SDs and explained the formation of the amorphous phase. In the stability test, virtually no time-dependent decrease was observed in either
in vitro
drug dissolution or drug content.
Conclusion:
SD formulation with n-HAp may be a promising approach for enhancing the dissolution and stability of TSIIA.
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Novel characterization of
Radix Angelicae Dahuricae
before and after the sulfur-fumigation process by combining high performance liquid chromatographic fingerprint and multi-ingredients determination
p. 338
Xiao Liu, Jingjing Liu, Hao Cai, Songlin Li, Xiaoqing Ma, Yajing Lou, Kunming Qin, Hongyue Guan, Baochang Cai
DOI
:10.4103/0973-1296.137376
PMID
:25210323
Background:
Harmful sulfur-fumigation processing method is abused during
Radix
Angelicae
Dahuricae
preparation. However, the analytical technique characterizing
Radix
Angelicae
Dahuricae
before and after the sulfur-fumigation process is absent.
Materials and Methods:
The high performance liquid chromatography (HPLC) technique was adopted to develop methods combining finger-print analysis and multi-ingredients simultaneous determination for quality evaluation of
Radix
Angelicae
Dahuricae
before and after the sulfur-fumigation process. The chromatographic fingerprint method was established for qualitative analysis coupled with statistical cluster analysis basing on Euclidean distance. Additionally, a determination method was developed for quantitative analysis, which was able to assay the concentrations of the major coumarins including imperatorin, isoimperatorin, xanthotoxin, xanthotoxol, isoimpinellin, oxypeucedanin, and bergapten in
Radix
Angelicae
Dahuricae
simultaneously. The separations of the two methods were both achieved on a Hypersil octadecylsilyl C
18
column (250 mm × 4.6 mm, 5 μm) at 35°C under different strategic gradient elution programs. The detection wavelength was set at 254 nm all the time. Method validation data indicated that the methods were both reliable and applicable. They were then used to assay different
Radix
Angelicae
Dahuricae
samples collected from good agricultural practice (GAP) bases and local herbal markets.
Results:
The successful application demonstrated that the combination of HPLC fingerprint and simultaneous quantification of multi-ingredients offers an efficient approach for quality evaluation of
Radix
Angelicae
Dahuricae
before and after the sulfur-fumigation process.
Conclusion:
In order to discriminate
Radix
Angelicae
Dahurica
e before and after the sulfur-fumigation process, oxypeucedanin, and xanthotoxol were the most sensitive biomarkers and should be determined.
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Phytochemical compositions of volatile oil from
Blumea balsamifera
and their biological activities
p. 346
Zhi-long Jiang, Yan Zhou, Wei-chen Ge, Ke Yuan
DOI
:10.4103/0973-1296.137377
PMID
:25210324
Background:
This research, extract compositions of volatile oil in
Blumea balsamifera
(Linn.) DC by steam distillation method. The gas chromatograph-mass spectrometer technology (GC-MS) was used to separate and determine chemical components of volatile oil. Meanwhile, the percentage of the chemical components was determined in the volatile oil by peak area normalization method. At the same time, we determine and evaluate antitumor, antibacterial, and antioxidation activities of chemical components of volatile oil in
Blumea balsamifera
of the aromatic plant.
Materials and Methods:
We screened the cytotoxicity of volatile oil in
Blumea balsamifera
by using prawn larva and by prawn-lethal bioactivity experiment. Use the slanting test tube method to evaluate the antibacterial activity of volatile oil in
Blumea balsamifera
for eight kinds of plant pathogenic fungi. Taking Trolox as the contrast, the research uses DPPH method to study the radical-scavenging function (IC
50
) of the volatile oil in
Blumea balsamifera
.
Results:
The results show that 42 kinds of compounds are separated from volatile oil of
Blumea balsamifera
. The appraised components take up 97.65% of total peak area. The volatile oil in
Blumea balsamifera
mainly contains sesquiterpenoids. The results also show that it has relatively strong activity of antitumor and anti-plant pathogenic fungi and some antioxidation activity.
Conclusion:
This research provided the reference data for further development of this natural resource, and at the same time, we understood more of the chemical components of volatile oil and bioactivity of this aromatic plant.
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A rapid identification of four medicinal chrysanthemum varieties with near infrared spectroscopy
p. 353
Bangxing Han, Hui Yan, Cunwu Chen, Houjun Yao, Jun Dai, Naifu Chen
DOI
:10.4103/0973-1296.137378
PMID
:25210325
Background:
For genuine medicinal material in Chinese herbs; the efficient, rapid, and precise identification is the focus and difficulty in the filed studying Chinese herbal medicines.
Chrysanthemum morifolium
as herbs has a long planting history in China, culturing high quality ones and different varieties. Different chrysanthemum varieties differ in quality, chemical composition, functions, and application. Therefore, chrysanthemum varieties in the market demands precise identification to provide reference for reasonable and correct application as genuine medicinal material.
Materials and Methods:
A total of 244 batches of chrysanthemum samples were randomly divided into calibration set (160 batches) and prediction set (84 batches). The near infrared diffuses reflectance spectra of chrysanthemum varieties were preprocessed by first order derivative (D1) and autoscaling and was built model with partial least squares (PLS).
Results:
In this study of four chrysanthemum varieties identification, the accuracy rates in calibration sets of Boju, Chuju, Hangju, and Gongju are respectively 100, 100, 98.65, and 96.67%; while the accuracy rates in prediction sets are 100% except for 99.1% of Hangju.
Conclusion:
The research results demonstrate that the qualitative analysis can be conducted by machine learning combined with near infrared spectroscopy (NIR), which provides a new method for rapid and noninvasive identification of chrysanthemum varieties.
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Simultaneous determination of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β -D-glucuronopyranside methyl ester and quercetin in effective part of
Polygonum Perfoliatum
L. using high performace liquid chromatography
p. 359
Dongsheng Fan, Yang Zhao, Xin Zhou, Xiaojian Gong, Chao Zhao
DOI
:10.4103/0973-1296.137379
PMID
:25210326
Objective:
In the present study, a high performance liquid chromatography (HPLC) coupled with photodiode array detection was developed for simultaneous quantitation of esculetin, quercetin-3-O-β-D-glucuronide, quercetin-3-O-β-D- glucuronopyranoside methyl ester and quercetin in
Polygonum perfoliatum
L.
Materials and Methods:
The chromatographic separations were performed on a reversed-phase C
18
column using a mobile phase composed of acetonitrile -0.5% aqueous acetic acid with gradient elution. The calibration curves for the analytes demonstrated good linearities within the investigated ranges. The satisfactory intra- and inter-day precision, repeatability and stability of the developed analytical method were shown in the method validation procedure. The recoveries of the established method ranged from 95.76 to 102.10% for all the analytes.
Results:
This proposed method was successfully applied for simultaneous quantification of the four compounds in effective part of
Polygonum perfoliatum
L. from different regions. Hierarchical clustering analysis (HCA) and principal components analysis (PCA) were performed to characterize and classify the samples based on the contents of the four compounds in
Polygonum perfoliatum
L.
Conclusion:
The established HPLC method combined with chemometric approaches was proven to be useful and efficient for quality control of
Polygonum perfoliatum
L.
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In vivo
anti-fatigue activity of sufu with fortification of isoflavones
p. 367
Yunxian Liu, Yun Zhou, Satoru Nirasawa, Eizo Tatsumi, Yongqiang Cheng, Lite Li
DOI
:10.4103/0973-1296.137380
PMID
:25210327
Background:
Sufu is a traditional Chinese fermented soybean food. Isoflavones are abundant in soybean and products incorporated with isoflavones exert many health benefits. The aim of this study was to investigate anti-fatigue effect of sufu fortified with isoflavones.
Materials and Methods:
In vivo
anti-fatigue activity of sufu with fortification of isoflavones (IF) was investigated in this study via exhaustive swimming test using ICR mice and determination of biochemical parameters. Factors relating to fatigue, including hepatic glycogen, blood lactic acid (BLA), blood urea nitrogen (BUN) were determined. The isoflavone composition in the IF sufu was also determined to explore the anti-fatigue activity of isoflavones.
Results:
During fermentation, isoflavone glucosides were converted into aglycones and both sufu with and without fortification of IF prolonged the exhaustive swimming time of ICR mice. Intake of sufu also increased the hepatic glycogen content, while it decreased the levels of both the blood lactic acid (BLA) and blood urea nitrogen (BUN) content. A dose-response relationship was observed in both exhaustive swimming and BLA clearance test, with medium dose (1%) fortification of IF revealing the highest activity.
Conclusion:
IF sufu could possess high anti-fatigue activity.
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Development and validation of an UPLC-ESI-MS/MS method for determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine in Evodiae Fructus
p. 374
Yang Zhao, Yunling Zhao, Xin Zhou, Xiaojian Gong
DOI
:10.4103/0973-1296.137381
PMID
:25210328
Objective:
Evodiae Fructus (EF), one of the most widely used traditional Chinese medicines, mainly consists of alkaloids, is widely used for the treatments of headache and gastrointestinal disorders. In this study, a sensitive and reliable UPLC-ESI-MS/MS method was developed for qualitative determination of dehydroevodiamine, limonin, evodiamine, and rutaecarpine.
Materials and Methods:
Chromatographic separations were accomplished on a Phenomenex Kinetex XB-C18 column (2.1 × 150 mm, 1.7 μm) by using a gradient elution profile with a mobile phase consisting of 0.5% formic acid in water (A) and acetonitrile (B). Detection was performed using multiple reactions monitoring mode under ESI in the positive ion mode.
Results:
The results showed good linearity over the investigated concentration ranges (
R
2
>0.9900) for the analytes. The limit of quantitations (LOQs) were 6.88 ng/mL for dehydroevodiamine, 18.6 ng/mL for limonin, 6.24 ng/mL for evodiamine, and 2.56 ng/mL for rutaecarpine, respectively. Intraday and interday precisions (relative standard deviations, %) were <5% and accuracies ranged from 92% to 106%.
Conclusion:
The validated method was successfully applied to assay the contents of the four compounds in EF samples from different regions, with which just 10 min was needed to analyze each sample.
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