scholarly journals Development and Validation of a Method for Quantification of Favipiravir as COVID-19 Management in Spiked Human Plasma

Molecules ◽  
2021 ◽  
Vol 26 (13) ◽  
pp. 3789
Author(s):  
Mohammad Hailat ◽  
Israa Al-Ani ◽  
Mohammed Hamad ◽  
Zainab Zakareia ◽  
Wael Abu Dayyih
Keyword(s):  
Human Plasma ◽  
Internal Standard ◽  
Weighting Factor ◽  
Hplc Method ◽  
Bioanalytical Method Validation ◽  
Spiked Human Plasma ◽  
Fda Guidance ◽  
Significant Difference ◽  
Us Fda ◽  
Carry Over

In the current work, a simple, economical, accurate, and precise HPLC method with UV detection was developed to quantify Favipiravir (FVIR) in spiked human plasma using acyclovir (ACVR) as an internal standard in the COVID-19 pandemic time. Both FVIR and ACVR were well separated and resolved on the C18 column using the mobile phase blend of methanol:acetonitrile:20 mM phosphate buffer (pH 3.1) in an isocratic mode flow rate of 1 mL/min with a proportion of 30:10:60 %, v/v/v. The detector wavelength was set at 242 nm. Maximum recovery of FVIR and ACVR from plasma was obtained with dichloromethane (DCM) as extracting solvent. The calibration curve was found to be linear in the range of 3.1–60.0 µg/mL with regression coefficient (r2) = 0.9976. However, with acceptable r2, the calibration data’s heteroscedasticity was observed, which was further reduced using weighted linear regression with weighting factor 1/x. Finally, the method was validated concerning sensitivity, accuracy (Inter and Intraday’s % RE and RSD were 0.28, 0.65 and 1.00, 0.12 respectively), precision, recovery (89.99%, 89.09%, and 90.81% for LQC, MQC, and HQC, respectively), stability (% RSD for 30-day were 3.04 and 1.71 for LQC and HQC, respectively at −20 °C), and carry-over US-FDA guidance for Bioanalytical Method Validation for researchers in the COVID-19 pandemic crisis. Furthermore, there was no significant difference for selectivity when evaluated at LLOQ concentration of 3 µg/mL of FVIR and relative to the blank.

scholarly journals DEVELOPMENT AND VALIDATION OF BIOANALYTICAL HPLC METHOD FOR SIMULTANEOUS ESTIMATION OF CILNIDIPINE AND NEBIVOLOL IN HUMAN PLASMA

2017 ◽  
Vol 9 (10) ◽  
pp. 253
Author(s):  
Aruna G. ◽  
Bharathi K ◽  
Kvsrg Prasad
Keyword(s):  
Human Plasma ◽  
Analytical Method ◽  
Internal Standard ◽  
Hplc Method ◽  
Pharmacokinetic Parameters ◽  
Simultaneous Estimation ◽  
Time Data ◽  
Bioanalytical Method Validation ◽  
Limit Of Quantitation ◽  
The Mean

Objective: To develop and validate a modified isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method for determination of cilnidipine and nebivolol in human plasma to be used for pharmacokinetic studies.Methods: The drug was extracted from plasma samples by direct protein precipitation technique using acetonitrile. Amlodipine was used as internal standard (IS). Samples were analyzed on BDS C18 column (250 x 4.6 mm, 5 µm), applying ortho phosphoric acid (0.1%): Acetonitrile, at a ratio of 45:55 v/v in isocratic mode as a mobile phase at a flow rate of 1 ml/min to attain adequate resolution. Separations were performed at room temperature and monitored at a wavelength of 260 nm after injection of 50μl samples into the HPLC system. The analytical method was validated according to FDA bioanalytical method validation guidance. The method was applied for pharmacokinetic study of cilnidipine and nebivolol tablets-10 mg and 5 mg were administered as a single dose to 6 healthy male rabbits under fasting condition. Twelve blood samples were withdrawn from each rabbit over 24 h periods. From the plasma concentration-time data of each individual, the pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were calculated.Results: A peak area was obtained for cilnidipine and nebivolol at 3.943 and 4.719 min retention time respectively. Linearity was established at a concentration range of 0.20-20 μg/ml (r2=0.999, n=8) for cilnidipine and 0.02-2 μg/ml (r2=0.999, n=8) for nebivolol. The lower limit of quantitation (LLOQ) was identifiable and reproducible at 0.2μg/ml for cilnidipine and 0.02 μg/ml for nebivolol. The coefficients of variation (%cv) of the intra-day and inter-day precision of cilnidipine at 600, 1000 and 1600ng/ml levels were found to be 6.90%, 6.19%, 5.22%; and 7.74%, 6.54%, 5.77%, respectively, which are lower than the accepted criteria limits (15-20 %). The mean recovery (%) cilnidipine at 600, 1000, and 1600ng/ml was found to be 101.03%, 99.27% and 104.87%, and for nebivolol 60, 100, and 160 ng/ml was found to be 106.13%, 107.03% and 98.06% respectively. Stability at different conditions and in autosampler was also established. The mean pharmacokinetic parameters; Cmax, Tmax, AUC0-t and AUC0-∞ were 6 ng/ml, 2 hr, 96.76 mg. hr/ml, 63.45 mg. hr/ml for cilnidipine and 5.8ng/ml, 2hr, 74.78 mg. hr/ml, 100.25 mg. hr/ml for nebivolol respectively.Conclusion: The present analytical method was found to be specific, sensitive, accurate and precise for quantification of cilnidipine and nebivolol in human plasma. It can be successively applied for pharmacokinetics, bioavailability and bioequivalence studies.

Validated RP-HPLC Method for Simultaneous Determination of Ribavirin, Sofosbuvir and Daclatasvir in Human Plasma: A Treatment Protocol Administered to HCV Patients in Egypt

2019 ◽  
Vol 57 (7) ◽  
pp. 636-643 ◽  
Author(s):  
Aya A Youssef ◽  
N Magdy ◽  
Lobna A Hussein ◽  
A M El-Kosasy
Keyword(s):  
Human Plasma ◽  
Method Validation ◽  
Treatment Protocol ◽  
Internal Standard ◽  
Gradient Elution ◽  
Hplc Method ◽  
Array Detector ◽  
Therapeutic Drug ◽  
Bioanalytical Method Validation ◽  
Rp Hplc

Abstract Egypt has the highest prevalence of hepatitis C virus (HCV) in the world thus it launched a national program for eliminating HCV aiming to treat 300,000 HCV patients per year. Three anti-HCV co-administered drugs; ribavirin (RBV), sofosbuvir (SF) daclatasvir (DAC) were simultaneously determined in human plasma by a validated, simple and sensitive RP-HPLC method using propyl paraben as an internal standard. Liquid–liquid extraction using ethyl acetate was used for samples extraction. Chromatographic separation was achieved on Scharlau® C18 column (250 × 4.6 mm2, 5 μm). Gradient elution was employed with a mobile phase mixture of water and acetonitrile at a flow rate 1 mL/min. UV detection using photodiode array detector was carried out at 207, 260 and 312 nm for RBV, SF and DAC, respectively. Method validation was performed according to the FDA guidelines for bioanalytical method validation. The calibration curves were linear over the ranges (0.5–80, 0.1–40 and 0.5–80 μg/mL) with average recoveries (100.64–108.28%, 98.48–105.91% and 97.68–101.38%) for RBV, SF and DAC, respectively. The intra-day and inter-day precision and accuracy results were within the acceptable limits. Stability assays revealed that the three studied analytes were stable during sample storage, preparation and injection. The method can be successfully applied in routine analysis of plasma of HCV patients treated with this combination therapy which aids in therapeutic drug monitoring and patients’ follow-up especially in Egypt and other developing countries fighting HCV.

scholarly journals DEVELOPMENT AND VALIDATION OF A SIMPLE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY–UV METHOD FOR THE DETERMINATION OF VINORELBINE - A CHEMOTHERAPEUTIC DRUG IN SPIKED HUMAN PLASMA

2019 ◽  
Vol 12 (1) ◽  
pp. 369
Author(s):  
Useni Reddy Mallu ◽  
Venkateswara Rao Anna ◽  
Bikshal Babu Kasimala
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Hplc Method ◽  
Pharmacokinetic Studies ◽  
Chemotherapeutic Drug ◽  
Spiked Human Plasma ◽  
Rp Hplc

Objective: Vinorelbine (VNRB) is a chemotherapeutic drug used to treat non-small cell lung cancer and breast cancer. Literature survey reveals that there are no reverse-phase high-performance liquid chromatography (RP-HPLC) methods reported for the estimation of VNRB in spiked human plasma. Hence, the present work aimed to develop a simple and efficient RP-HPLC method for the estimation of VNRB in human plasma.Methods: Specimen preparation for the measurement of VNRB was performed through liquid-liquid extraction using methanol as extracting solvent and reconstructed with mobile phase. Paclitaxel (PCTX) was used as internal standard. HPLC method was optimized and validated as per the US FDA bioanalytical guidelines. VNRB and internal standard were separated on Kromasil® C18 (250×4.6 mm; id 5 μ) using acetate buffer (pH=5.9) and methanol in the ratio of 85:15 (v/v) at 1 ml/min flow rate. Eluted compounds were recorded using UV detector at 235 nm.Results: The retention time of PCTX and internal standard was found to be 4.3 and 9.0 min, respectively. The analytical measuring ranges were found to be 5–750 ng/ml (r2>0.9998). The method was found to be simple, accurate, precise, and stable and there is no interference of plasma matric components.Conclusion: The described HPLC method allows for the measurement of total and free PCTX in both plasma and cord blood and can utilize for the estimation of drug in pharmacokinetic studies.

DEVELOPMENT OF LIQUID-LIQUID EXTRACTION PROCEDURE AND ITS APPLICATION FOR QUANTIFICATION OF ERLOTINIB IN SPIKED HUMAN PLASMA USING RP-HPLC WITH UV DETECTION

INDIAN DRUGS ◽  
2016 ◽  
Vol 53 (06) ◽  
pp. 46-50
Author(s):  
A Faizee ◽  
◽  
S. S Sonawane ◽  
A. S. Patil ◽  
S. J Kshirsagar ◽  
...  
Keyword(s):  
Human Plasma ◽  
Ammonium Acetate ◽  
Extraction Procedure ◽  
Internal Standard ◽  
Hplc Method ◽  
Liquid Extraction ◽  
Calibration Model ◽  
Spiked Human Plasma ◽  
Liquid Liquid Extraction ◽  
Rp Hplc

A simple, rapid and accurate RP-HPLC method was developed and validated for the quantification of Erlotinib in spiked human plasma using liquid-liquid extraction. Sufficient recovery was obtained when drug and internal standard (Nabumetone) were extracted using ethyl acetate and 1N NaOH. Chromatographic separation was performed on C18 Phenomenex Hyperclone column (250 × 4.6 mm, 5 μm) using mobile phase acetonitrile: 20 mM ammonium acetate buffer pH 4.6 (60:40%,V/V). Flow rate was kept constant at 1 mL/min and detection was carried out at 331 nm. Calibration curve was found to be linear in the range of 100-3200 ng/mL. During the calibration experiments, it was found that heteroscedasticity can be minimized using weighted regression calibration model with weighing factor of 1/x2.

scholarly journals DEVELOPMENT AND VALIDATION OF A SIMPLE HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY–UV METHOD FOR THE DETERMINATION OF VINORELBINE - A CHEMOTHERAPEUTIC DRUG IN SPIKED HUMAN PLASMA

2019 ◽  
Vol 12 (1) ◽  
pp. 369 ◽  
Author(s):  
Useni Reddy Mallu ◽  
Venkateswara Rao Anna ◽  
Bikshal Babu Kasimala
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Hplc Method ◽  
Pharmacokinetic Studies ◽  
Chemotherapeutic Drug ◽  
Spiked Human Plasma ◽  
Rp Hplc

Objective: Vinorelbine (VNRB) is a chemotherapeutic drug used to treat non-small cell lung cancer and breast cancer. Literature survey reveals that there are no reverse-phase high-performance liquid chromatography (RP-HPLC) methods reported for the estimation of VNRB in spiked human plasma. Hence, the present work aimed to develop a simple and efficient RP-HPLC method for the estimation of VNRB in human plasma.Methods: Specimen preparation for the measurement of VNRB was performed through liquid-liquid extraction using methanol as extracting solvent and reconstructed with mobile phase. Paclitaxel (PCTX) was used as internal standard. HPLC method was optimized and validated as per the US FDA bioanalytical guidelines. VNRB and internal standard were separated on Kromasil® C18 (250×4.6 mm; id 5 μ) using acetate buffer (pH=5.9) and methanol in the ratio of 85:15 (v/v) at 1 ml/min flow rate. Eluted compounds were recorded using UV detector at 235 nm.Results: The retention time of PCTX and internal standard was found to be 4.3 and 9.0 min, respectively. The analytical measuring ranges were found to be 5–750 ng/ml (r2>0.9998). The method was found to be simple, accurate, precise, and stable and there is no interference of plasma matric components.Conclusion: The described HPLC method allows for the measurement of total and free PCTX in both plasma and cord blood and can utilize for the estimation of drug in pharmacokinetic studies.

scholarly journals Method Development and Validation for Determination of Febuxostat from Spiked Human Plasma Using RP-HPLC with UV Detection

2014 ◽  
Vol 2014 ◽  
pp. 1-5 ◽  
Author(s):  
Monita Gide ◽  
Pankaj Sharma ◽  
Ravindra Saudagar ◽  
Birendra Shrivastava
Keyword(s):  
Human Plasma ◽  
High Performance ◽  
Method Development ◽  
Internal Standard ◽  
Hplc Method ◽  
Uv Detection ◽  
Least Square ◽  
Spiked Human Plasma ◽  
Rp Hplc ◽  
Development And Validation

A rapid, simple, selective, and specific reverse phase high performance liquid chromatography (RP-HPLC) method with UV detection (315 nm) was developed and validated for estimation of febuxostat from spiked human plasma. The analyte and internal standard (diclofenac) were extracted using LLE with diethyl ether. The chromatographic separation was performed on Shodex C-18-4E (5 μm; 250×4.6 mm) with a mobile phase comprised of methanol : acetate buffer pH 4, 20 mM (90 : 10 v/v), at a flow rate of 1 mL/min. Febuxostat was well resolved from plasma constituents and internal standard. The calibration curve was linear in the range of 250–8000 ng/mL. The heteroscedasticity was minimized by using weighted least square regression with weighing factor of 1/x. The intraday and interday %RSD was less than 15. Results of recovery studies prove the extraction efficiency. Stability data indicated that febuxostat was stable in plasma after three freeze thaw cycles and upon storage at −20°C for 30 days.

An HPLC method for simultaneous quantification of sunitinib and its active metabolite, SU12662, using hydrophilic interaction chromatography principle

Bioanalysis ◽  
2020 ◽  
Vol 12 (2) ◽  
pp. 75-85 ◽  
Author(s):  
Murari Gurjar ◽  
Parsshava Mehta ◽  
Jyoti Sharma ◽  
Sneha Patil ◽  
Preeti Kulkarni ◽  
...  
Keyword(s):  
Human Plasma ◽  
Active Metabolite ◽  
Internal Standard ◽  
Hplc Method ◽  
Therapeutic Drug ◽  
Hydrophilic Interaction ◽  
Reliable Determination ◽  
The Us ◽  
Us Fda

Aim: To develop a sensitive HPLC method for the quantitation of sunitinib (SU) and its active metabolite N-desethyl-sunitinib (SU12662) in human plasma. Materials & methods: The analytes were extracted from 500 μl of plasma using liquid–liquid extraction followed by protein precipitation. Chromatographic separation of two analytes and internal standard, vandetenib, was achieved on a hydrophilic interaction liquid chromatography analytical column using a gradient program. Calibration curves were linear over the range of 10–250 ng/ml for both SU and SU12662. The method was validated according to the US FDA guidelines for bioanalytical methods. Accuracy of the method at 10 ng/ml for SU and SU12662 was 8.7 and 6.7%, respectively, and precision was 10.18% and 17.3%, respectively. Conclusion: This method allows a specific, sensitive and reliable determination of SU and SU12662 in human plasma in a single analytical run which makes it useful for therapeutic drug monitoring.

scholarly journals High Performance Liquid Chromatoqraphic Method for the Simultaneous Determination of Labetalol and Hydrochlorothiazide in Tablets and Spiked Human Plasma

2004 ◽  
Vol 72 (2) ◽  
pp. 143-155 ◽  
Author(s):  
M. Sultan ◽  
H. Abdine ◽  
N. Zoman ◽  
F. Belal
Keyword(s):  
Simultaneous Determination ◽  
Human Plasma ◽  
High Performance ◽  
Internal Standard ◽  
Reversed Phase ◽  
Hplc Method ◽  
Spiked Human Plasma ◽  
Quantitation Limit

A reversed-phase HPLC method with spectrophotometric detection was developed for the simultaneous determination of labetalol (LBT) and hydrochloro-thiazide (HCD). The chromatographic separation was performed using a Microbondapak C18 column (4.6 i.d. x 250 nm) and paracetamol as internal standard. A mobile phase consisting of 0.05 M phosphate buffer/acetonitrile of pH 4 (7:3) at a flow rate of 0.7 ml/min was used. The detection was affected spectrophotornetrically at 302 nm. The working concentration range was 0.3–10 µg/ml with detection limits of 0.05 µg/ml for both drugs. The lower quantitation limit was 0.25 µg/ml in the two cases. The method was successfully applied to tablets, the % recoveries were 99.45 ± 0.68 for LBT and 99.79 ± 0.75 for HCD. The method was extended to the in-vitro determination in spiked human plasma. The % recoveries were 91.12 ± 0.33 for LBT and 91.37 ± 0.40 for HCD. The interday and intraday precision and accuracy were evaluated in plasma by calculating the % RSD (n=5) and the % error and were found to be in the ranges of 1.18–4.1% and 0.38–0.36% for both drugs, respectively.

Validation of a Reversed‐Phase HPLC Method for the Analysis of Sildenafil Citrate in Pharmaceutical Preparations and in Spiked Human Plasma

2005 ◽  
Vol 28 (10) ◽  
pp. 1539-1548 ◽  
Author(s):  
Bülent Ergun ◽  
Ahmet Saracoglu ◽  
Sinem Ilgin ◽  
Zeki Atkosar ◽  
Kevser Kircali ◽  
...  
Keyword(s):  
Human Plasma ◽  
Pharmaceutical Preparations ◽  
Reversed Phase ◽  
Hplc Method ◽  
Sildenafil Citrate ◽  
Reversed Phase Hplc ◽  
Spiked Human Plasma
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