Full factorial experimental design for development and validation of a RP-HPLC method for estimation of letrozole in nanoformulations

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Abstract

Background: Letrozole (LTZ) is a potent aromatase inhibitor which blocks estrogen synthesis in post-menopausal women. Suitable analytical methods are required to quantify letrozole in bulk and its nanoformulations. Hence a sensitive and robust reverse-phase high performance liquid chromatographic method (RP-HPLC) was developed for the estimation of LTZ. Methods: The method was developed using Kinetex C18 column (250 mm×4.6 mm; 5μ) and mobile phase comprising acetonitrile (ACN): acetate buffer (pH 4.5) mixture (50:50% v/v) was used to affect the chromatographic separation at 0.8 mL/min flow rate. The responses were measured at 240 nm. Force degradation studies were done by exposing LTZ to acid- and alkali-induced hydrolysis, oxidation (H2 O2), thermal and photolysis. Two-level factorial design was used to validate the method as per ICH Q2 (R1) using Design-Expert® software. The effects of independent variables on flow rate, pH, acetonitrile content and column temperature were recorded as responses. Results: Method was linear over the concentration range of 5-2500 ng/mL with a correlation coefficient (R2) of 1.000. Force degradation studies revealed that LTZ was stable to all stress agents except the basic conditions. The inter-day precision results were reproducible with relative standard deviation (% RSD) of 0.078. The peak area RSD and mean recovery of LTZ was found to be <1.0% and 99-102% in drug solution and <1.0% and 98-106% in nanoformulations respectively. Deliberate changes in independent chromatographic parameters analyzed using analysis of variance (ANOVA) indicated that the model was significant (p<0.0001). Conclusion: The developed analytical method was successfully utilized to quantify LTZ in bulk and its nanoformulations.

Original languageEnglish
Pages (from-to)320-330
Number of pages11
JournalCurrent Pharmaceutical Analysis
Volume14
Issue number3
DOIs
Publication statusPublished - 01-05-2018

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letrozole
Design of experiments
Research Design
Liquids
Flow rate
Software Design
Degradation
Aromatase Inhibitors
Photolysis
Alkalies
Analysis of variance (ANOVA)
Hydrolysis
Analysis of Variance

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Biochemistry
  • Molecular Medicine
  • Pharmaceutical Science

Cite this

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title = "Full factorial experimental design for development and validation of a RP-HPLC method for estimation of letrozole in nanoformulations",
abstract = "Background: Letrozole (LTZ) is a potent aromatase inhibitor which blocks estrogen synthesis in post-menopausal women. Suitable analytical methods are required to quantify letrozole in bulk and its nanoformulations. Hence a sensitive and robust reverse-phase high performance liquid chromatographic method (RP-HPLC) was developed for the estimation of LTZ. Methods: The method was developed using Kinetex C18 column (250 mm×4.6 mm; 5μ) and mobile phase comprising acetonitrile (ACN): acetate buffer (pH 4.5) mixture (50:50{\%} v/v) was used to affect the chromatographic separation at 0.8 mL/min flow rate. The responses were measured at 240 nm. Force degradation studies were done by exposing LTZ to acid- and alkali-induced hydrolysis, oxidation (H2 O2), thermal and photolysis. Two-level factorial design was used to validate the method as per ICH Q2 (R1) using Design-Expert{\circledR} software. The effects of independent variables on flow rate, pH, acetonitrile content and column temperature were recorded as responses. Results: Method was linear over the concentration range of 5-2500 ng/mL with a correlation coefficient (R2) of 1.000. Force degradation studies revealed that LTZ was stable to all stress agents except the basic conditions. The inter-day precision results were reproducible with relative standard deviation ({\%} RSD) of 0.078. The peak area RSD and mean recovery of LTZ was found to be <1.0{\%} and 99-102{\%} in drug solution and <1.0{\%} and 98-106{\%} in nanoformulations respectively. Deliberate changes in independent chromatographic parameters analyzed using analysis of variance (ANOVA) indicated that the model was significant (p<0.0001). Conclusion: The developed analytical method was successfully utilized to quantify LTZ in bulk and its nanoformulations.",
author = "Hegde, {Aswathi R.} and Managuli, {Renuka S.} and Anup Naha and Koteshwara, {Kunnatur B.} and Reddy, {Meka Sreenivasa} and Srinivas Mutalik",
year = "2018",
month = "5",
day = "1",
doi = "10.2174/1573412913666171006152604",
language = "English",
volume = "14",
pages = "320--330",
journal = "Current Pharmaceutical Analysis",
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T1 - Full factorial experimental design for development and validation of a RP-HPLC method for estimation of letrozole in nanoformulations

AU - Hegde, Aswathi R.

AU - Managuli, Renuka S.

AU - Naha, Anup

AU - Koteshwara, Kunnatur B.

AU - Reddy, Meka Sreenivasa

AU - Mutalik, Srinivas

PY - 2018/5/1

Y1 - 2018/5/1

N2 - Background: Letrozole (LTZ) is a potent aromatase inhibitor which blocks estrogen synthesis in post-menopausal women. Suitable analytical methods are required to quantify letrozole in bulk and its nanoformulations. Hence a sensitive and robust reverse-phase high performance liquid chromatographic method (RP-HPLC) was developed for the estimation of LTZ. Methods: The method was developed using Kinetex C18 column (250 mm×4.6 mm; 5μ) and mobile phase comprising acetonitrile (ACN): acetate buffer (pH 4.5) mixture (50:50% v/v) was used to affect the chromatographic separation at 0.8 mL/min flow rate. The responses were measured at 240 nm. Force degradation studies were done by exposing LTZ to acid- and alkali-induced hydrolysis, oxidation (H2 O2), thermal and photolysis. Two-level factorial design was used to validate the method as per ICH Q2 (R1) using Design-Expert® software. The effects of independent variables on flow rate, pH, acetonitrile content and column temperature were recorded as responses. Results: Method was linear over the concentration range of 5-2500 ng/mL with a correlation coefficient (R2) of 1.000. Force degradation studies revealed that LTZ was stable to all stress agents except the basic conditions. The inter-day precision results were reproducible with relative standard deviation (% RSD) of 0.078. The peak area RSD and mean recovery of LTZ was found to be <1.0% and 99-102% in drug solution and <1.0% and 98-106% in nanoformulations respectively. Deliberate changes in independent chromatographic parameters analyzed using analysis of variance (ANOVA) indicated that the model was significant (p<0.0001). Conclusion: The developed analytical method was successfully utilized to quantify LTZ in bulk and its nanoformulations.

AB - Background: Letrozole (LTZ) is a potent aromatase inhibitor which blocks estrogen synthesis in post-menopausal women. Suitable analytical methods are required to quantify letrozole in bulk and its nanoformulations. Hence a sensitive and robust reverse-phase high performance liquid chromatographic method (RP-HPLC) was developed for the estimation of LTZ. Methods: The method was developed using Kinetex C18 column (250 mm×4.6 mm; 5μ) and mobile phase comprising acetonitrile (ACN): acetate buffer (pH 4.5) mixture (50:50% v/v) was used to affect the chromatographic separation at 0.8 mL/min flow rate. The responses were measured at 240 nm. Force degradation studies were done by exposing LTZ to acid- and alkali-induced hydrolysis, oxidation (H2 O2), thermal and photolysis. Two-level factorial design was used to validate the method as per ICH Q2 (R1) using Design-Expert® software. The effects of independent variables on flow rate, pH, acetonitrile content and column temperature were recorded as responses. Results: Method was linear over the concentration range of 5-2500 ng/mL with a correlation coefficient (R2) of 1.000. Force degradation studies revealed that LTZ was stable to all stress agents except the basic conditions. The inter-day precision results were reproducible with relative standard deviation (% RSD) of 0.078. The peak area RSD and mean recovery of LTZ was found to be <1.0% and 99-102% in drug solution and <1.0% and 98-106% in nanoformulations respectively. Deliberate changes in independent chromatographic parameters analyzed using analysis of variance (ANOVA) indicated that the model was significant (p<0.0001). Conclusion: The developed analytical method was successfully utilized to quantify LTZ in bulk and its nanoformulations.

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