HPLC method development/validation and skin diffusion study of caffeine, methyl paraben and butyl paraben as skin–diffusing model drugs

The focus of this research was to develop and validate a suitable HPLC method, which allows simultaneous determination of three proposed skin model penetrants to investigate the percutaneous diffusion behavior of their combination: caffeine, methyl paraben and butyl paraben. These penetrants were selected because they represent a wide range of lipophilicities. This model highlights the effect of combining penetrants of different molecular properties on their diffusion behavior through skin. The proposed method employed a gradient system that was systematically optimized for separation and quantification of the penetrants. The effect of the stationary phase (C18, C4 and cyano (CN)) was assessed with CN proven to be superior in terms of peak shape, retentivity and dynamic linear range. Significant differences in retention time, peak broadening, and quantifiability between different stationary phases could be demonstrated. The method was validated as per ICH guidelines Q2 (R1) with a satisfactory outcome. The method was successfully applied for real diffusion experiments, and revealed notable differences between the individual penetrants and their ternary mixture on transdermal permeation. The method could potentially be extended to determine these analytes in other related skin permeation investigations.

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Analytical and preparative separation and isolation of functionalized fullerenes by conventional HPLC stationary phases: method development and column screening

RSC Advances ◽

10.1039/d0ra02814b ◽

2020 ◽

Vol 10 (33) ◽

pp. 19211-19218

Author(s):

Merve Ergun Dönmez ◽

Helena Grennberg

Keyword(s):

Method Development ◽

Stationary Phases ◽

Hplc Method ◽

Complex Reaction ◽

Preparative Separation ◽

Isolation And Purification ◽

Functionalized Fullerenes

Isolation and purification of functionalized fullerenes from often complex reaction mixtures is challenging. Here, a simple and efficient HPLC method is presented.

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Simultaneous Determination of Saponins and a Flavonoid from Aerial Parts of Zygophyllum coccineum L.

Journal of AOAC International ◽

10.5740/jaoacint.11-216 ◽

2012 ◽

Vol 95 (3) ◽

pp. 757-762

Author(s):

Elham Amin ◽

Yan-Hong Wang ◽

Bharathi Avula ◽

Seham S El-Hawary ◽

Magda M Fathy ◽

...

Keyword(s):

Simultaneous Determination ◽

Relevant Literature ◽

Hplc Method ◽

Desert Plant ◽

Gradient System ◽

Alcohol Extract ◽

Triterpenoid Saponins ◽

Wide Range ◽

Acquity Uplc

Abstract Triterpenoid saponins are a class of glycosides with a wide range of bioactivities, which make them interesting research candidates. Zygophyllum coccineum is an Egyptian desert plant rich in triterpenoid saponins. Reviewing the relevant literature, no data concerning the HPLC or ultra-performance LC (UPLC) analysis of Zygophyllum content were found. This paper presents two methods, HPLC-UV and UPLC-UV-evaporative light scattering detector (ELSD)/MS, for the simultaneous determination of 10 compounds in the alcohol extract of Z. coccineum. The HPLC method uses a C18 column and water–acetonitrile (both containing 0.1% trifluoroacetic acid) gradient system. The separation was achieved within 32 min. The developed UPLC method simultaneously detects and quantifies the 10 compounds using an Acquity UPLC BEH Shield RP18 column and reagent alcohol–acetonitrile (80/20, v/v) and water (both containing 0.5% formic acid) gradient system within 14 min with UV, ELS, and MS detectors. The methods were used to analyze another species, Z. simplex, and results revealed a great variation between the secondary metabolite pattern of both species.

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Chromatographic Characterization and Method Development for Determination of Levetiracetam in Saliva: Application to Correlation with Plasma Levels

Journal of Analytical Methods in Chemistry ◽

10.1155/2017/7846742 ◽

2017 ◽

Vol 2017 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

Imad I. Hamdan ◽

Mervat Alsous ◽

Amira Taher Masri

Keyword(s):

High Performance ◽

Method Development ◽

Stationary Phases ◽

High Performance Liquid Chromatographic ◽

Hplc Method ◽

Therapeutic Drug ◽

Epileptic Children ◽

Isocratic Hplc ◽

Liquid Chromatographic

Levetiracetam (LVT) is a widely used antiepileptic drug (AED). A less invasive sampling method for therapeutic drug monitoring (TDM) would be very useful particularly for children. Saliva has been shown as an adequate sample for TDM of some AEDs. Due to the high hydrophilicity of LVT its separation on common stationary phases is quite a challenge so that previous methods for determination of LVT in saliva employed either gradient high performance liquid chromatographic (HPLC) system or mass spectrometer as a detector. In this study the retention behavior of LVT on some common stationary phases was examined, with C8 being the most retentive. A simple isocratic HPLC method that is based on simple protein precipitation was developed and validated for the determination of LVT in saliva. The method was applied to a sample group of epileptic children for the purpose of assessing potential correlation with plasma LVT levels and to investigate patient’s compliance. The results confirmed a reasonable correlation between plasma and salivary levels of LVT (R = 0.9) which supports the use of saliva for TDM of LVT. The study also revealed a significant percentage of epileptic patients having LVT levels below the estimated therapeutic range.

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HPLC METHOD DEVELOPMENT AND VALIDATION FOR AZITHRO-MYCIN IN ORAL SUSPENSION

Journal of Applied Pharmaceutical Sciences and Research ◽

10.31069/japsr.v2i1.2 ◽

2019 ◽

pp. 7-12

Author(s):

Alok Pratap Singh ◽

Iti Chauhan ◽

Snigdha Bhardwaj ◽

Praveen Gaur ◽

S Sadish Kumar ◽

...

Keyword(s):

Method Development ◽

Limit Of Detection ◽

Hplc Method ◽

Uv Detector ◽

Limit Of Quantification ◽

Relative Standard ◽

Validation Parameters ◽

Wide Range ◽

Tract Infections ◽

Development And Validation

Introduction: Azithro-mycin a semi-synthetic, azalide congener of erythro-mycin indicated in the treatment of respiratory tract infections. Various methods available for determination of Azithro-mycin, but HPLC are most versatile one. Objective: The present study is based on the development and validation of a rapid, simple high performance liquid chromatography (HPLC) method equipped with UV detector for quantitative analysis of Azithro-mycin (AZN) in suspension. Material and methods: The Method was performed by using Hypersil BDS-C18 (250 mm × 4.6 mm i.d.) column MS-II, with an isocratic mobile phase of methanol, acetonitrile and phosphate buffer pH 8 (60:30:10; v/v) with run time 15 minutes. The determinations were performed at a flow rate of 1.0ml/min, and UV detector set at 212 nm. Result and Discussion: The method was found to be specific with relative standard deviation (RSD) less than 2.09%. The method showed accuracy with RSD less than 1.34% and precision in repeatability with RSD less than 1.42%. The method was found to be linear over a wide range of concentration from 1.0 to 50.0 μg/mL (R2 = .995). Limit of detection and limit of quantification were found to be 14.40 ng/mL and 43.66 ng/mL respectively. Conclusion: It was advantageous to use UV detector over other methods employing electrochemical, photodiode array etc. as the detector, because of cheap and easy availability. The developed method fulfilled all validation parameters as per ICH and can be successfully applied to quantify percent drug content in marketed oral Azithro-mycin suspension.

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Method Development and Validation of Degradation Studies of Lenalidomide by RP-HPLC

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00744 ◽

2021 ◽

pp. 4281-4286

Author(s):

Punna Venkateshwarlu ◽

Mehul M. Patel

Keyword(s):

Retention Time ◽

Mobile Phase ◽

Method Development ◽

Hplc Method ◽

Mobile Phase Flow Rate ◽

Pharmaceutical Dosage Forms ◽

Rp Hplc ◽

System Suitability ◽

Wide Range

A simple, accurate, RP HPLC method was developed by this study determination of lenalidomide. This method is developed by Shimadzu LC -2010 HT by using C18 (250 X 4.6 X mm X 5µ) column in solvents Phosphate buffer: Acetonitrile (55:45) v/v as mobile phase and the temperature was maintained at 25°C. The mobile phase flow rate 1ml/min was pumped and sample wavelength was detected at 242nm by ultraviolet -visible spectrophotometer. The retention time was found 2.5 min. The number of theoretical plates and tailing factor for lenalidomide was observed 16199.817 (NLT 2000) and 1.128 (NMT 2). The method was validated for analytical standards such as linearity, accuracy, precision, system suitability and robustness. LOD and LOQ values obtained from regression of lenalidomide 0.058 and 0.174µg/ml. The regression equation of validated method for lenalidomide is Y=5223x+183075. In wide range of 25 to 150 (µg/ml) the linearity was observed. The method was validated and a recovery study indicates accuracy of this method. The Retention time less compared to established methods. The method was validated by determining its accuracy, precision and system suitability. The results of the study showed that the proposed RP-HPLC method is simple, rapid, precise and accurate, which is useful for the routine determination of Lenalidomide in bulk drug and in its pharmaceutical dosage forms.

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Method Development and Validation for Estimation of related Substances in Tilorone Dihydrochloride using RP¬-HPLC

Research Journal of Pharmacy and Technology ◽

10.52711/0974-360x.2021.00577 ◽

2021 ◽

pp. 3319-3324

Author(s):

M. Zeba Baktiyar ◽

B. Mohammed Ishaq ◽

Siva Sanker Reddy L ◽

Sreenivasulu M.

Keyword(s):

Mobile Phase ◽

Method Development ◽

Hplc Method ◽

Column Temperature ◽

Related Substances ◽

Rp Hplc ◽

Ich Guidelines ◽

Wide Range ◽

Method Development And Validation ◽

Development And Validation

A simple, precise and reproducible RP-HPLC method was developed for the estimation of related substances in tilorone dihydrochloride. Quantification was performed using a Zorbax SB-phenyl column (150 × 4.6mm, 5µ) with mobile phase A: 20mM potassium dihydro phosphate + 2ml of triethylamine, pH 2.30 and mobile phase B: acetonitrile, methanol and water 60: 20: 20% v/v. A gradient program was followed with a run time of 55 minutes at a flow rate of 1.0 ml/min. The column temperature was maintained at 40°C, the injection volume was 10 µl and the detection was performed at 269nm using a PDA detector. The retention time of Tilorone dihydrochloride was found to be 10.36 minutes. The proposed method has been validated according to the ICH guidelines for Linearity, Precision, Accuracy, LOD, and LOQ. The method was linear from 0.157 - 3.934μg/ml for standard, 0.153-3.820μg/ml and 0.166 - 4.140μg/ml for impurities, TLHC01 and TLHC02 respectively. The impurities TLHC01 and TLHC02 have been mapped in all stress conditions. The LOD and LOQ of TLHC01 were found at 1.757μg/ml and 5.857μg/ml and 1.919μg/ml and 6.396μg/ml respectively for TLHC02 respectively. Statistical analysis showed that the method was precision, reproducible, selective, specific and accurate for the analysis of Tilorone dihydrochloride and its impurities. The wide range of linearity, sensitivity, precision, short retention times and simple mobile phase have shown that the method is suitable for the routine quantification of mass impurities of tilorone hydrochloride and its dosage pharmaceutical forms with high precision and accuracy.

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Method Development and Validation of Degradation Studies of Favipiravir by RP-HPLC

Journal of Pharmaceutical Research International ◽

10.9734/jpri/2021/v33i42a32399 ◽

2021 ◽

pp. 220-228

Author(s):

. Shyamala ◽

Dongamanti Ashok

Keyword(s):

Mobile Phase ◽

Method Development ◽

Hplc Method ◽

Mobile Phase Flow Rate ◽

Rp Hplc ◽

Wide Range ◽

Method Development And Validation ◽

Development And Validation ◽

Analytical Standards ◽

Theoretical Plates

RP HPLC method was developed by this study estimation of favipiravir. This method is developed by Shimadzu LC -2010 HT by using a C18 (250 X 4.6 X mm X 5µ) column in solvents Water(OPA)+ACN (60:40)v/v as mobile phase and the temperature was maintained at 25°C. The mobile phase flow rate of 1ml/min was pumped and sample wavelength was detected at 324 nm by ultraviolet -visible spectrophotometer. The Rt was found 4.453 min. The number of theoretical plates and tailing factor for favipiravir was observed 82651(NLT 2000) and 1.265 (NMT 2). The method was validated for analytical standards such as linearity, accuracy, precision, LOD, LOQ and robustness. LOD and LOQ values were calculated from regression of favipiravir 1.26 and 3.83 µg/ml.The regression equation of validated method for favipiravir is Y=253.5x+1881.In a wide range of 4 to 20 (µg/ml) the linearity was observed. Degradation methods were also performed.

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Rp-HPLC Determination of Quercetin in a Novel D-α-Tocopherol Polyethylene Glycol 1000 Succinate Based SNEDDS Formulation: Pharmacokinetics in Rat Plasma

Molecules ◽

10.3390/molecules26051435 ◽

2021 ◽

Vol 26 (5) ◽

pp. 1435

Author(s):

Osama A. A. Ahmed ◽

Hany M. El-Bassossy ◽

Heba M. El-Sayed ◽

Soad S. Abd El-Hay

Keyword(s):

Polyethylene Glycol ◽

Limit Of Detection ◽

Rat Plasma ◽

Hplc Method ◽

Pharmacokinetic Studies ◽

Gradient System ◽

Rp Hplc ◽

Polyethylene Glycol 200 ◽

Wide Range ◽

Polyethylene Glycol 1000

Despite its proven efficacy in diverse metabolic disorders, quercetin (QU) for clinical use is still limited because of its low bioavailability. D-α-Tocopherol polyethylene glycol 1000 succinate (TPGS) is approved as a safe pharmaceutical adjuvant with marked antioxidant and anti-inflammatory activities. In the current study, several QU-loaded self-nanoemulsifying drug delivery systems (SNEDDS) were investigated to improve QU bioavailability. A reversed phase high performance liquid chromatography (RP-HPLC) method was developed, for the first time, as a simple and sensitive technique for pharmacokinetic studies of QU in the presence of TPGS SNEDDS formula in rat plasma. The analyses were performed on a Xterra C18 column (4.6 × 100 mm, 5 µm) and UV detection at 280 nm. The analytes were separated by a gradient system of methanol and phosphate buffer of pH 3. The developed RP-HPLC method showed low limit of detection (LODs) of 7.65 and 22.09 ng/mL and LOQs of 23.19 and 66.96 ng/mL for QU and TPGS, respectively, which allowed their determination in real rat plasma samples. The method was linear over a wide range, (30–10,000) and (100–10,000) ng/mL for QU and TPGS, respectively. The selected SNEDDS formula, containing 50% w/w TPGS, 30% polyethylene glycol 200 (PEG 200), and 20% w/w pumpkin seed oil (PSO), showed a globule size of 320 nm and −28.6 mV zeta potential. Results of the pharmacokinetic studies showed 149.8% improvement in bioavailability of QU in SNEDDS relative to its suspension. The developed HPLC method proved to be simple and sensitive for QU and TPGS simultaneous determination in rat plasma after oral administration of the new SNEDDS formula.

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