Quantitative Analysis of Eugenol in Clove Extract by a Validated HPLC Method

Abstract Clove (Eugenia caryophyllata) is a well-known medicinal plant used for diarrhea, digestive disorders, or in antiseptics in Korea. Eugenol is the main active ingredient of clove and has been chosen as a marker compound for the chemical evaluation or QC of clove. This paper reports the development and validation of an HPLC-diode array detection (DAD) method for the determination of eugenol in clove. HPLC separation was accomplished on an XTerra RP18 column (250 4.6 mm id, 5m) with an isocratic mobile phase of 60 methanol and DAD at 280 nm. Calibration graphs were linear with very good correlation coefficients (r2 > 0.9999) from 12.5 to 1000 ng/mL. The LOD was 0.81 and the LOQ was 2.47 ng/mL. The method showed good intraday precision (RSD 0.080.27) and interday precision (RSD 0.321.19). The method was applied to the analysis of eugenol from clove cultivated in various countries (Indonesia, Singapore, and China). Quantitative analysis of the 15 clove samples showed that the content of eugenol varied significantly, ranging from 163 to 1049 ppb. The method of determination of eugenol by HPLC is accurate to evaluate the quality and safety assurance of clove, based on the results of this study.

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Development and Validation of Stability Indicating RP-HPLC Method for Determination of Enzalutamide

International Journal of Pharmaceutical Sciences and Nanotechnology ◽

10.37285/ijpsn.2019.12.5.3 ◽

2019 ◽

Vol 12 (5) ◽

pp. 4635-4645

Author(s):

Pankaj Padmakar Nerkar ◽

Sameer Ansari ◽

Shailesh Chalikwar

Keyword(s):

Ammonium Acetate ◽

Correlation Coefficients ◽

Reversed Phase ◽

Hplc Method ◽

Stability Indicating ◽

Stability Indicating Method ◽

Rp Hplc ◽

Ich Guidelines ◽

Development And Validation

A simple, isocratic, and accurate reversed phase HPLC method was developed for the quantitative determination of enzalutamide. The chromatographic separation was achieved on an Qualisil BDS C18 (250 mm x 4.6mm, 5 μm) column using methanol: ammonium acetate buffer pH 4.2 adjusted with glacial acetic acid: (60:40, v/v) as a mobile phase, at a flow rate of 1 ml/min and detection at 236nm. The linear range for enzalutamide were 2.0 to 10 μg/mL was obtained with correlation coefficients ≥ 0.998. The retention time was found to be 6.30min. Enzalutamide was subjected to stress conditions hydrolysis (acid, base) oxidation, photolysis and thermal degradation and the stressed samples were analysed by the developed method. The method was validated for the precision, accuracy, linearity and robustness. The developed stability indicating method for enzalutamide was validated as per ICH guidelines.

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Development and validation of SPE-HPLC method for the determination of carbamazepine and its metabolites carbamazepine epoxide and carbamazepine trans-diol in plasma

Journal of the Serbian Chemical Society ◽

10.2298/jsc120106084d ◽

2012 ◽

Vol 77 (10) ◽

pp. 1423-1436 ◽

Cited By ~ 8

Author(s):

Predrag Dzodic ◽

Ljiljana Zivanovic ◽

Ana Protic ◽

Ivana Ivanovic ◽

Radmila Velickovic-Radovanovic ◽

...

Keyword(s):

Plasma Sample ◽

Correlation Coefficients ◽

Hplc Method ◽

Optimal Conditions ◽

Relative Standard ◽

Epileptic Patients ◽

Oasis Hlb ◽

Development And Validation ◽

Carbamazepine Epoxide

SPE-HPLC method has been developed and validated for rapid analysis of carbamazepine and its two metabolites carbamazepine epoxide and carbamazepine trans-diol in human plasma. The analysis was performed using C18 Bakerbond-BDC analytical column (250 mm x 4.6 mm i.d., particle size 5 ?m). The optimal conditions for the separation were established with the mobile phase acetonitrile - 10 mM phosphate buffer, pH 7.0 (30:70, v/v) at the flow rate of 1.5 mL min-1, temperature 35?C, and UV detection at 210 nm. Total run time was about 8 minutes. SPE procedure for extraction of the analytes from plasma sample was developed using Oasis HLB cartridges and subsequently eluate was injected into the HPLC system for analysis. Afterwards, SPE-HPLC method was subjected to validation. Linearity was obtained over the concentration range of 0.2-25 ?g/mL for carbamazepine, carbamazepine epoxide and carbamazepine trans-diol with correlation coefficients higher than 0.995. The method showed good intra-day and inter-day precision with relative standard deviation below 7.96%, while accuracy ranged from 92.09% to 108.5% for all analytes. Finally, the method was successfully applied to analysis of plasma samples of epileptic patients in monotherapy and polytherapy. [Acknowledgments. Projekat Ministarstva nauke Republike Srbije, br. OI 172033].

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HPLC identification and determination of myricetin, quercetin, kaempferol and total flavonoids in herbal drugs

Macedonian Pharmaceutical Bulletin ◽

10.33320/maced.pharm.bull.2002.48.005 ◽

2003 ◽

Vol 48 ◽

pp. 25-30 ◽

Cited By ~ 4

Author(s):

Svetlana Kulevanova ◽

Marina Stefova ◽

Tatjana Kadifkova Panovska ◽

Trajce Stafilov

Keyword(s):

Quantitative Analysis ◽

Hplc Method ◽

Total Flavonoids ◽

Herbal Drugs ◽

Column Temperature ◽

Total Flavonoids Content ◽

Array Detection ◽

Hydrolysis Of ◽

Uv Range

A new and rapid HPLC method for identification and determination of myricetin, quercetin, kaempferol and total flavonoids in ten herbal drugs of Macedonian origin is presented. Preparation of samples (Uvae ursi folim, Pruni spinosae flos, Sambuci flos, Betulae folim, Primulae flos, Herniariae herba, Centaurii herba, Tiliae flos, Robiniae pseudoacaciae flos, Bursae pastoris herba) included hydrolysis of glycosides and extraction of total aglycones with ethyl acetate. HPLC analysis with UV-diode array detection was carried out on RP C18 column, using 5% acetic acid and acetonitrile in agradient elution mode and column temperature of 30 o C. The monitoring of the elution is performed in the whole UV-range and the acquisition of data for quantitative analysis at 367 nm. Screening of the extracts showed presence of quercetin in nine, kaempferol in seven and myricetin in only one sample. The quantitative analysis showed that the content of quercetin ranged from 0.026-0.506 % (m/m), while for kaempferol it was from traces to 1.246 %. Uvaeursi folium and Pruni spinosae flos were rich in content of quercetin (0.482 % and 0.506 %, respectively), while Pruni spinosae flos and Robiniae pseudoaccaciae flos contained the highest amounts of kaempferol (1.246 % and 0.892 %, respectively). Myricetin was identified and determined only in Betulae folium (0.102 %). The content of total flavonoids in the investigated samples expressed in terms of quercetin ranged from 0.040 to 1.680 %. The proposed HPLC method is convenient for use in routine analysis of myricetin, quercetin and kaempferol, as well as for estimation of total flavonoids content in herbal drugs.

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Development and Validation of a RP-HPLC Method for the Simultaneous Determination of LevamisoleHCl and Oxyclozanideand its Application in the Assay of Veterinary Bolus Formulations

Current Pharmaceutical Analysis ◽

10.2174/1573412916666200211101633 ◽

2020 ◽

Vol 16 ◽

Author(s):

Kemal Hussien Seid ◽

Tarekegn Berhanu ◽

Kaleab Asres ◽

Ayenew Ashenef

Keyword(s):

High Performance Liquid Chromatography ◽

Liquid Chromatography ◽

High Performance ◽

Correlation Coefficients ◽

Hplc Method ◽

Reverse Phase ◽

Rp Hplc ◽

Average Accuracy ◽

Development And Validation

Introduction: A reverse-phase high performance liquid chromatography (RP-HPLC) method was developed and validated for the simultaneous analysis of two drugs, levamisole hydrochloride(LH) and oxyclozanide(OX), in co-formulations for veterinary use. Materials and Methods: The new HPLC method was validated per ICH and other guidelines. A C18 column was used with agradient program; eluent A was an equal mixture of methanol and acetonitrile, and eluent B a 25 mM phosphate buffer at pH 7.0 containing 30 mM sodium decanesulfonate andtriethylamine(50:50:1 v/v)then pH adjusted to 7.0 with H3PO4 [51:49 v/v] .The detection wavelength was set at 220 nm.For the final gradient program, the retention times were 8.2(for LH)and 13.6(for OX) minutes respectively at flow rate of 1 ml/min over 20 minute run time. Results: The method wasprecise, specific and robust.The correlation coefficients, R2 were 0.9998 and 0.9999 for LH and OX respectively in the ranges of 5 – 280 µg / mL.The percent y-intercepts and percent residual standard deviations were 1.6%/0.4% and 1.4%/1.0% for LH and OX, respectively. The LOD and LOQ of the method were 0.21 µg / mL and 0.62 µg / mL for LHand 0.06 µg / mL and 0.18 µg / mL for OX. The method has average accuracy of 100.5% for LH and 101.1% for OX when tested on veterinary bolus formulations, and the samples couldbe stored under typical lab conditions for about 7 days without significant degradation. Conclusion: This HPLC method is suitable forassayinglevamisole hydrochloride and oxyclozanide simultaneously from veterinary formulations.

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