scholarly journals Application of HPLC-DAD for In Vitro Investigation of Acetylcholinesterase Inhibition Activity of Selected Isoquinoline Alkaloids from Sanguinaria canadensis Extracts

Molecules ◽  
2021 ◽  
Vol 26 (1) ◽  
pp. 230
Author(s):  
Tomasz Tuzimski ◽  
Anna Petruczynik
Keyword(s):  
Plant Extracts ◽  
High Performance ◽  
Acetylcholinesterase Activity ◽  
Hplc Method ◽  
Acetylcholinesterase Inhibition ◽  
Isoquinoline Alkaloids ◽  
Activity Inhibition ◽  
Sanguinaria Canadensis ◽  
In Vitro Investigation

Isoquinoline alkaloids may have a wide range of pharmacological activities. Some of them have acetylcholinesterase activity inhibition. Nowadays, neurodegenerative disorders such as Alzheimer’s disease have become a serious public health problem. Searching for new effective compounds with inhibited acetylcholinesterase activity is one of the most significant challenges of modern scientific research. The aim of this study was the in vitro investigation of acetylcholinesterase activity inhibition of extracts obtained from Sanguinaria canadensis collected before, during and after flowering. The acetylcholinesterase activity inhibition of these extracts has not been previously tested. The aim was also to quantify selected alkaloids in the investigated extracts by high performance liquid chromatography (HPLC). The analyses of alkaloid content were performed using HPLC in reversed phase (RP) mode using Polar RP column and mobile phase containing acetonitrile, water and ionic liquid (IL). The acetylcholinesterase activity inhibition of the tested plant extracts and respective alkaloid standards were examined using high performance liquid chromatography with diode-array detector (HPLC-DAD) for the quantification of 5-thio-2-nitro-benzoic acid, which is the product of the reaction between the thiocholine (product of the hydrolysis of acetylthiocholine reaction) with Ellman reagent. The application of the HPLC method allowed for elimination of absorption of interfering components, for example, alkaloids such as sanguinarine and berberine. It is revealed that the HPLC method can be successfully used for the evaluation of the acetylcholinesterase inhibitory activity in samples such as plant extracts, especially those containing colored components adsorbing at wavelength in the range 405–412 nm. The acetylcholinesterase inhibition activity synergy of pairs of alkaloid standards and mixture of all investigated alkaloids was also determined. Most investigated alkaloids and all Sanguinaria canadensis extracts exhibited very high acetylcholinesterase activity inhibition. IC50 values obtained for alkaloid standards were from 0.36 for berberine to 23.13 µg/mL for protopine and from 61.24 to 89.14 µg/mL for Sanguinaria canadensis extracts. Our investigations demonstrated that these plant extracts can be recommended for further in vivo experiments to confirm their acetylcholinesterase activity inhibition.

scholarly journals Badanie wpływu naturalnych ekstraktów roślinnych na izolowane wierzchołki wzrostów pedów w hodowli in vitro [Investigation of the influence of natural plant extracts on isolated generative apices cultivated in vitro]

2015 ◽  
Vol 26 (1) ◽  
pp. 5-15
Author(s):  
E. Kuta ◽  
C. Ryka
Keyword(s):  
Plant Extracts ◽  
Plant Material ◽  
Mineral Nutrient ◽  
Careful Selection ◽  
Natural Plant ◽  
In Vitro Investigation ◽  
Coconut Meal ◽  
Selection Of ◽  
Spinach Leaves

The aim of the present paper Was to test the value of various plant extracts (yeast barm, mature walnut embryos, coconut meal, young spinach leaves and tomato juice) used as the complementary elements of White's mineral nutrient for the culture <i>in vitro</i> of generative apices of winter wheat, rape and wetch. The results showed that the generative apices of investigated planits had various nutritive requirements which point to the necessity of a careful selection of a specific nutrient for particular plant material.

In Vitro Effects of Light on Serum Bilirubin Subfractions Measured by High-Performance Liquid Chromatography: Comparison with Four Routine Methods

1992 ◽  
Vol 38 (10) ◽  
pp. 2124-2129 ◽  
Author(s):  
H Ihara ◽  
H Nakamura ◽  
Y Aoki ◽  
T Aoki ◽  
M Yoshida
Keyword(s):  
High Performance ◽  
Total Bilirubin ◽  
Serum Bilirubin ◽  
Accurate Determination ◽  
Hplc Method ◽  
Direct Bilirubin ◽  
Spectrometric Method ◽  
Indirect Bilirubin ◽  
In Vitro Effects

Abstract The effects of light on serum bilirubin subfractions in vitro were investigated by HPLC and four routine methods for bilirubin analysis. By HPLC, the rate of photodegradation of unconjugated bilirubin (Bu) was nearly twice that of monoconjugated bilirubin (mBc) and threefold that of diconjugated bilirubin (dBc); delta bilirubin (Bd) was most stable against photoirradiation. In the diazo method, the rate of photodegradation of direct bilirubin was almost the same as that of the sum of mBc, dBc, and Bd determined by the HPLC method. However, the rate of photodegradation of indirect bilirubin was significantly lower (P &lt; 0.001) than that obtained by HPLC, because approximately 30% of the bilirubin photoproducts reacted with the diazo reagent as indirect bilirubin. The rate of photodegradation of total bilirubin determined by the direct spectrometric method was lower than that determined by the diazo method, but equal to that of the total peak areas of HPLC. In the Ektachem method, bilirubin photoproducts affected total bilirubin negligibly, and Bc and Bu positively, so that the value of Bd decreased. In the bilirubin oxidase method, bilirubin photoproducts were oxidized enzymatically by both the total and direct bilirubin reagents. We re-emphasize the importance of shielding serum from light to avoid generating bilirubin photoproducts that interfere with the accurate determination of serum bilirubin subfractions. We also recommend HPLC analysis as a standard method for bilirubin measurement.

The In Vitro Degradation Kinetics of Polyurethane Prodrug Materials

2011 ◽  
Vol 399-401 ◽  
pp. 1067-1070
Author(s):  
Chun Yan Li ◽  
Cong Cong Hu ◽  
Zhi Guo Wen ◽  
Sheng Xiong Dong
Keyword(s):  
Drug Release ◽  
High Performance ◽  
Degradation Kinetics ◽  
Hplc Method ◽  
In Vitro Degradation ◽  
Erosion Mechanism ◽  
First Order ◽  
First Order Kinetics ◽  
Kinetics Of

The method of high performance liquid chromatography (HPLC) is established to determine the content of antibacterial agent — ciprofloxacin (CF) in the degradation solution of ciprofloxacin-polyurethane (CFPU) and investigate the in vitro degradation kinetics by plotting and fitting the cumulative release curves to inspect the effects of different medium and different concentrations on drug release. The results showed that the HPLC method is accurate, reliable and simple. The drug-release of CFPU was bioresponsive and could be accorded with first order kinetics. It was observed that CF was released from CFPU by a combination of diffusion and erosion mechanism, mainly in the manner of diffusion in the absence of infection while erosion mechanism in the presence of infection.

scholarly journals RP-HPLC Method Development and Validation for Simultaneous Estimation of Clarithromycin and Paracetamol

2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Sadana Gangishetty ◽  
Surajpal Verma
Keyword(s):  
High Performance ◽  
Method Development ◽  
Hplc Method ◽  
Simultaneous Estimation ◽  
Uv Detector ◽  
Rp Hplc ◽  
Ich Guidelines ◽  
Hplc Method Development ◽  
Development And Validation

The present work describes a simple, rapid, and reproducible reverse phase high performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of clarithromycin (CLA) and paracetamol (PCM). C18 column (Kromasil ODS, 5 µm, 250 × 4.6 mm) and a mobile phase containing phosphate buffer (0.05 M) along with 1-octane sulphonic acid sodium salt monohydrate (0.005 M) adjusted to pH 3.2: acetonitrile (50 : 50 v/v) mixture was used for the separation and quantification. The flow rate was 1.0 mL/min and the eluents were detected by UV detector at 205 nm. The retention times were found to be 2.21 and 3.73 mins, respectively. The developed method was validated according to ICH guidelines Q2 (R1) and found to be linear within the range of 75–175 µg/mL for both drugs. The developed method was applied successfully for assay of clarithromycin and paracetamol in their combined in-house developed dosage forms and in vitro dissolution studies.

scholarly journals HPLC analysis, antimicrobial and antioxidant activities of Daphne cneorum L.

2012 ◽  
Vol 66 (5) ◽  
pp. 709-716 ◽  
Author(s):  
Nedeljko Manojlovic ◽  
Pavle Maskovic ◽  
Perica Vasiljevic ◽  
Ratomir Jelic ◽  
Marina Juskovic ◽  
...  
Keyword(s):  
High Performance ◽  
Antimicrobial Agents ◽  
Inhibitory Concentration ◽  
Total Phenolics ◽  
Antioxidant Activities ◽  
High Performance Liquid Chromatographic ◽  
Hplc Method ◽  
Potential Source ◽  
Liquid Chromatographic

The present study describes in vitro antimicrobial and antioxidant activities of the methanol extracts which were obtained from the leaves and twigs of the plant Daphne cneorum L. The antimicrobial activity of these extracts was tested against human pathogenic microorganisms using a minimum inhibitory concentration (MIC) values. Total phenolics and flavonoids, as well as the antioxidant activity of the extracts were determined. The two tested extracts showed good antimicrobial and antioxidant activities. The results of a high performance liquid chromatographic (HPLC) method showed that 7,8-dihydroxycoumarin is one of the most abundant secondary metabolite in the tested extracts. The results of this study clearly indicated that the extracts of D. cneorum could be used as a potential source of natural antioxidant and antimicrobial agents.

scholarly journals RP-HPLC Method Development and Validation of Synthesized Codrug in Combination with Indomethacin, Paracetamol, and Famotidine

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Mohyeddin Assali ◽  
Murad Abualhasan ◽  
Nihal Zohud ◽  
Noura Ghazal
Keyword(s):  
High Performance ◽  
Method Development ◽  
Pharmaceutical Formulations ◽  
Hplc Method ◽  
Active Ingredients ◽  
Reverse Phase ◽  
Rp Hplc ◽  
Relative Standard ◽  
Hplc Method Development

Background. Indomethacin is considered a potent nonsteroidal anti-inflammatory drug that could be combined with Paracetamol to have superior and synergist activity to manage pain and inflammation. To reduce the gastric side effect, they could be combined with Famotidine. Methodology. A codrug of Indomethacin and Paracetamol was synthesized and combined in solution with Famotidine. The quantification of the pharmaceutically active ingredients is pivotal in the development of pharmaceutical formulations. Therefore, a novel reverse-phase high-performance liquid chromatography (RP-HPLC) method was developed and validated according to the International Council for Harmonization (ICH) Q2R1 guidelines. A reverse phase C18 column with a mobile phase acetonitrile: sodium acetate buffer 60 : 40 at a flow rate of 1.4 mL/min and pH 5 was utilized. Results. The developed method showed good separation of the four tested drugs with a linear range of 0.01–0.1 mg/mL (R2 > 0.99). The LODs for FAM, PAR, IND, and codrug were 3.076 × 10−9, 3.868 × 10−10, 1.066 × 10−9, and 4.402 × 10−9 mg/mL respectively. While the LOQs were 9.322 × 10−9, 1.172 × 10−10, 3.232 × 10−9, and 1.334 × 10−8 mg/mL, respectively. Furthermore, the method was precise, accurate, selective, and robust with values of relative standard deviation (RSD) less than 2%. Moreover, the developed method was applied to study the in vitro hydrolysis and conversion of codrug into Indomethacin and Paracetamol. Conclusion. The codrug of Indomethacin and Paracetamol was successfully synthesized for the first time. Moreover, the developed analytical method, to our knowledge, is the first of its kind to simultaneously quantify four solutions containing the following active ingredients of codrug, Indomethacin, Paracetamol, and Famotidine mixture with added pharmaceutical inactive ingredients in one HPLC run.

scholarly journals Validation and Application of a New Reversed Phase HPLC Method forIn VitroDissolution Studies of Rabeprazole Sodium in Delayed-Release Tablets

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Md. Saddam Nawaz
Keyword(s):  
High Performance ◽  
Method Development ◽  
High Performance Liquid Chromatographic ◽  
Reversed Phase ◽  
Hplc Method ◽  
Array Detector ◽  
Relative Standard ◽  
Quality Control Tool ◽  
Rabeprazole Sodium

The purpose of this study was to develop and validate a new reversed phase high performance liquid chromatographic (RP-HPLC) method to quantifyin vitrodissolution assay of rabeprazole sodium in pharmaceutical tablet dosage form. Method development was performed on C 18,100×4.6 mm ID, and 10 μm particle size column, and injection volume was 20 μL using a diode array detector (DAD) to monitor the detection at 280 nm. The mobile phase consisted of buffer: acetonitrile at a ratio of 60 : 40 (v/v), and the flow rate was maintained at 1.0 mL/min. The method was validated in terms of suitability, linearity, specificity, accuracy, precision, stability, and sensitivity. Linearity was observed over the range of concentration 0.05–12.0 μg/mL, and the correlation coefficient was found excellent >0.999. The method was specific with respect to rabeprazole sodium, and the peak purity was found 99.99%. The method was precise and had relative standard deviations (RSD) less than 2%. Accuracy was found in the range of 99.9 to 101.9%. The method was robust in different variable conditions and reproducible. This proposed fast, reliable, cost-effective method can be used as quality control tool for the estimation of rabeprazole sodium in routine dissolution test analysis.

scholarly journals ANALYTICAL METHOD DEVELOPMENT, VALIDATION AND STABILITY STUDIES BY RP-HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF ANDROGRAPHOLIDE AND CURCUMIN IN CO-ENCAPSULATED NANOSTRUCTURED LIPID CARRIER DRUG DELIVERY SYSTEM

2021 ◽  
pp. 73-86
Author(s):  
ASIT KUMAR DE ◽  
TANMOY BERA
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
High Performance ◽  
Method Development ◽  
Drug Loading ◽  
Hplc Method ◽  
Simultaneous Estimation ◽  
Nanostructured Lipid Carrier ◽  
Rp Hplc

Objective: The current study aims to boost the bioavailability criteria of two natural bioactive compounds, andrographolide and curcumin by their combination in nanostructured lipid carrier (NLC) and also to develop a straightforward reverse-phase high-performance liquid chromatography (RP-HPLC) method to validate, quantify of andrographolide and curcumin simultaneously in novel NLC formulation. Methods: The reliable chromatographic separation was executed by using a column of Phenomenex octadecylsilane (C18) at 35 °C column oven temperature using a mobile phase of 0.02 M potassium dihydrogen orthophosphate (KH2PO4) salt solution of pH 3.0 as a buffer and acetonitrile in 50: 50 v/v fixed ratio and 1.5 ml/min flow rate of with 20 μl injection load. The detection was carried out at 240 nm isosbestic wavelength employing a photodiode array (PDA) detector. Results: Andrographolide and curcumin were eluted at 2.4 and 4.9 min, respectively. Quantification and linearity were achieved for both drugs at the 10-140 μg/ml range. The method is specified as the presence of excipients utilized in the formulation failed to interfere with the estimation of andrographolide and curcumin. The developed method was successfully utilized to work out the drug loading efficiency and in vitro drug release study of those drugs in NLC formulation and also for the estimation of those drugs from rat plasma. Conclusion: The developed high-performance liquid chromatography (HPLC) method may be utilized in the future estimation of andrographolide and curcumin simultaneously in NLC and other nanoformulations both in vitro and in vivo.

scholarly journals Effect of isoquinoline alkaloids from two Hippeastrum species on in vitro acetylcholinesterase activity

Phytomedicine ◽  
2010 ◽  
Vol 17 (8-9) ◽  
pp. 698-701 ◽  
Author(s):  
L.B. Pagliosa ◽  
S.C. Monteiro ◽  
K.B. Silva ◽  
J.P. de Andrade ◽  
J. Dutilh ◽  
...  
Keyword(s):  
Acetylcholinesterase Activity ◽  
Isoquinoline Alkaloids
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