Original Article
Comparison of UV spectrophotometry and high
performance liquid chromatography methods
for the determination of repaglinide in tablets
Abstract
Ǽȱ–Ž‘˜œȱ Ž›ŽȱŽŸŽ•˜™Žȱ˜›ȱŽŽ›–’—Š’˜—ȱ˜ȱ
ȱŠ“žœŽȱ˜ȱřǯśȱ ’‘ȱ˜›‘˜™‘˜œ™‘˜›’ŒȱŠŒ’ǼȱŠȱŠȱ̘ ȱ›ŠŽȱ˜ȱŗǯŖȱ–•Ȧ–’—ǯȱ
Š›–˜—’£Š’˜—ȱ ǻ
Ǽȱ
Seema M. Dhole,
ȱ–Ž‘˜œǰȱ›Žœ™ŽŒ’ŸŽ•¢ǯȱ›ŽŒ’œ’˜—ȱǻƖǯǯȱǀȱŗǯśŖǼȱŠ—ȱ–ŽŠ—ȱ›ŽŒ˜ŸŽ›’Žœȱ Ž›Žȱ
Pramod B. Khedekar1,
Nikhil D. Amnerkar
ȱ–Ž‘˜ȱ ‘’Œ‘ȱœ‘˜ œȱŠŒŒž›ŠŒ¢ȱ˜ȱ‘Žȱ–Ž‘˜œǯȱConclusion: The
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Access this article online
Website: www.phmethods.org
DOI: 10.4103/2229-4708.103875
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Pharmaceutical Methods | July-December 2012 | Vol 3 | Issue 2
Background: Repaglinide is a miglitinide class of antidiabetic drug used for the
treatment of type 2 diabetes mellitus. A fast and reliable method for the determination
of repaglinide was highly desirable to support formulation screening and quality
control. Objective: UV spectrophotometric and reversed-phase high performance
•’šž’ȱŒ‘›˜–Š˜›Š™‘¢ȱǻȬ
repaglinide in the tablet dosage form. Materials and Methods: The UV spectrum
recorded between 200 400 nm using methanol as solvent and the wavelength 241 nm
was selected for the determination of repaglinide. RP-HPLC analysis was carried out
žœ’—ȱ’•Ž—ȱȬŗŞȱǻŘǼȱŒ˜•ž–—ȱŠ—ȱ–˜‹’•Žȱ™‘ŠœŽȱŒ˜–™˜œŽȱ˜ȱ–Ž‘Š—˜•ȱŠ—ȱ ŠŽ›ȱ
ǻŞŖDZŘŖȱŸȦŸǰȱ™
Š›Š–ŽŽ›œȱœžŒ‘ȱŠœȱ•’—ŽŠ›’¢ǰȱ™›ŽŒ’œ’˜—ǰȱŠŒŒž›ŠŒ¢ǰȱ›ŽŒ˜ŸŽ›¢ǰȱœ™ŽŒ’ęŒ’¢ȱŠ—ȱ›žŽ—Žœœȱ
Š›Žȱ œž’Žȱ Šœȱ ›Ž™˜›Žȱ ’—ȱ ‘Žȱ —Ž›—Š’˜—Š•ȱ ˜—Ž›Ž—ŒŽȱ ˜—ȱ
guidelines. Results:ȱ‘ŽȱŽŸŽ•˜™Žȱ–Ž‘˜œȱ’••žœ›ŠŽȱŽ¡ŒŽ••Ž—ȱ•’—ŽŠ›’¢ȱǻ›2 ǁȱŖǯşşşǼȱ
’—ȱ‘ŽȱŒ˜—ŒŽ—›Š’˜—ȱ›Š—Žȱ˜ȱśȬřŖȱΐȦ–•ȱŠ—ȱśȬśŖȱΐȦ–•ȱ˜›ȱȱœ™ŽŒ›˜™‘˜˜–Ž›’Œȱ
Š—ȱ
˜ž—ȱ’—ȱ‘Žȱ›Š—Žȱ˜ȱşşǯŜřȬŗŖŖǯŚśƖȱ˜›ȱȱœ™ŽŒ›˜™‘˜˜–Ž›’Œȱ–Ž‘˜ȱŠ—ȱşşǯŝŗȬ
ŗŖŖǯŘśƖȱ˜›ȱ
developed methods were found to be reliable, simple, fast, accurate and successfully
used for the quality control of repaglinide as a bulk drug and in pharmaceutical
formulations.
Key words: Method validation, quantitative analysis, repaglinide, RP-HPLC
INTRODUCTION
Type 2 diabetes is a long-term metabolic disorder wherein the body becomes
›Žœ’œŠ—ȱ ˜ȱ ‘Žȱ ŽěŽŒœȱ ˜ȱ ’—œž•’—ǰȱ Šȱ ‘˜›–˜—Žȱ ‘Šȱ ›Žž•ŠŽœȱ œžŠ›ȱ Š‹œ˜›™’˜—ǯȱ
›ŽŠ–Ž—ȱ˜ȱ¢™ŽȬŘȱ’Š‹ŽŽœȱǻ—˜—’—œž•’—ȬŽ™Ž—Ž—Ǽȱ’œȱ—˜ ȱ™˜œœ’‹•Žȱ ’‘ȱ˜›Š••¢ȱ
Š–’—’œŽ›Žȱ ‘¢™˜•¢ŒŽ–’Œȱ ŠŽ—œȱ ‘Šȱ ‘Ž•™ȱ ˜ȱ ›ŽžŒŽȱ ‹•˜˜ȱ œžŠ›ȱ •ŽŸŽ•œǯȹ[1]
Ž™Š•’—’Žȱ ’œȱ Šȱ ŒŠ›‹˜–˜¡¢•–Ž‘¢•ȱ ‹Ž—£˜’Œȱ ŠŒ’ȱ Ž›’ŸŠ’ŸŽǰȱ Š•œ˜ȱ ”—˜ —ȱ Šœȱ
ŘȬŽ‘˜¡¢ȬŚȬǽŘȬǽǽřȬ–Ž‘¢•ȬŗȬǽŘȬ•Ȭ™’™Ž›’’—¢•Ǽ™‘Ž—¢•Ǿ‹ž¢•ǾŠ–’—˜ǾȬŘȬ˜¡¢Ž‘¢•Ǿǯȱ
The chemical structure of repaglinide is shown in Figure 1. Repaglinide is a
meglitinide antidiabetic drug used for the treatment of type-2 diabetes mellitus
and it lowers blood glucose by stimulating the release of insulin from the pancreas.
It achieves this by closing ATP-dependent potassium channels in the membrane
˜ȱ‘ŽȱΆȬŒŽ••œǯǽŘǰřǾ
ŽŒ‘—˜•˜¢ȱ Š—ȱ œŒ’Ž—’ęŒȱ ™›˜›Žœœȱ ‘Šœȱ •Žȱ ˜ȱ ‘Žȱ ŽŸŽ•˜™–Ž—ȱ ˜ȱ —ž–Ž›˜žœȱ
œ¢—‘Ž’Œȱ›žœǯȱ‘Žȱ’—Œ›ŽŠœŽȱž’•’£Š’˜—ȱ˜ȱ‘ŽœŽȱŠ—’’Š‹Ž’Œȱ›žœȱŽ–Š—œȱ‘Žȱ
development of new and alternative methods to successfully determine these
›žœȱ’—ȱ›Š ȱ–ŠŽ›’Š•ǰȱ™‘Š›–ŠŒŽž’ŒŠ•ȱ˜œŠŽȱ˜›–ȱŠ—ȱ’—ȱ‘Žȱ‹’˜•˜’ŒŠ•ȱ̞’œǯ
Extensive literature survey reveals that previous studies have reported the
determination of repaglinide employing UV and visible spectrophotomeric,[4-6]
68
 Dhole, et al.: Spectrophotometric and HPLC method for repaglinide
ȱŠ“žœŽȱ˜ȱřǯśȱ ’‘ȱ˜›‘˜™‘˜œ™‘˜›’Œȱ
Figure 1: Repaglinide
spectrofluorimetric methods, [7] HPLC, [8,9]ȱ Ȧ
MS focusing mainly in its quantitation in plasma.[10]
These reported methods were mainly focused on the
Š—Š•¢œ’œȱ˜ȱ‘Žȱ›žȱ‹¢ȱŒ˜•˜›’–Ž›¢ȱŠ—ȱžœŽœȱ˜ȱ‹žěŽ›ȱ
in mobile phase for HPLC method. The purpose of
this study was to develop simple, fast, economical and
validated analytical methods to quantify repaglinide
in tablets using HPLC and UV spectrophotometry.
Validation of the developed methods was done as
™Ž›ȱ —Ž›—Š’˜—Š•ȱ ˜—Ž›Ž—ŒŽȱ ˜—ȱ
Š›–˜—’£Š’˜—ȱ concentrations to obtain the calibration graph.
ǻ
Ǽȱ ž’Ž•’—Žœǯ[11] The results obtained by these
methods were statistically compared using analysis of
variance. In addition the reliability and feasibility of
these methods were evaluated, focusing on rountine
quality control analysis.
MATERIALS AND METHODS
Materials
Repaglinide, reference standard was obtained as
Šȱ Ž—Ž›˜žœȱ ’Ğȱ œŠ–™•Žȱ ›˜–ȱ ȱ Š‹ǯȱ Ÿǯȱ ǯǰȱ
Mumbai, India. Eurepa tablets labelled to contain
›Ž™Š•’—’Žȱ Řȱ –ǰȱ –Š—žŠŒž›Žȱ ‹¢ȱ Ȧȱ ˜››Ž—ȱ
‘Š›–ŠŒŽž’ŒŠ•ȱ ǯǰȱ Š’ȱ ǻ
ǯǯǼǰȱ —’Šǰȱ  Ž›Žȱ for UV spectrophotometric and with mobile phase for
purchased from local market. All the chemicals used
were of AR and HPLC grade, obtained from E. Merck,
India.
Instrumentation and optimization conditions
UV spectrophotometric analyses were carried
˜žȱ ˜—ȱ Šȱ ‘’–Š£žȱ ŗŝŖŖȱ ˜ž‹•Žȱ ‹ŽŠ–ȱ Ȭ’œȱ
œ™ŽŒ›˜™‘˜˜–ŽŽ›ǰȱ  ’‘ȱ ŗǯŖȬŒ–ȱ šžŠ›£ȱ ŒŽ••œǯȱ ‘Žȱ
wavelength of 241 nm was selected for the quantitation
of repaglinide and the measurements were obtained
against methanol as a blank.
The HPLC analysis were carried out on Agilent
1120 Compact LC system composed of binary
Pharmaceutical Methods | July-December 2012 | Vol 3 | Issue 2
69
™ž–™ǰȱ –Š—žŠ•ȱ ’—“ŽŒ˜›ǰȱ ȱ ŽŽŒ˜›ȱ Š—ȱ £Œ‘›˜–Žȱ
•’Ž˜–™ŠŒȱœ˜Ğ Š›Žǯȱ‘ŽȱŒ˜•ž–—ȱžœŽȱ Šœȱ’•Ž—ȱ
ȬŗŞȱǻŘśŖȱ––ȱƼȱŚǯŜȱ––ȱ’ǯǯǰȱśȱΐ–ȱ™Š›’Œ•Žȱœ’£ŽǼȱŠ—ȱ
the mobile phase consisted of methanol and water
ǻŞŖDZŘŖȱŸȦŸǰȱ™
ŠŒ’Ǽȱ Šȱ Šȱ •˜ ȱ ›ŠŽȱ ˜ȱ ŗǯŖȱ –•Ȧ–’—ǯȱ ŽŽŒ’˜—ȱ  Šœȱ
performed on at 241 nm.
Preparation of standard solution
‘ŽȱœŠ—Š›ȱœ˜Œ”ȱœ˜•ž’˜—ȱ˜ȱ›Ž™Š•’—’ŽȱŗŖŖŖȱΐȦȹ–•ȱ
was prepared in methanol. For spectrophotometric
method, further dilutions of aliquots of standard stock
solution were carried out with methanol to reach the
Œ˜—ŒŽ—›Š’˜—ȱ›Š—ŽȱśȬřŖȱΐȦ–•ȱ˜›ȱ›Ž™Š•’—’Žǯȱ‘Žȱ
absorbance of series of solutions was measured at 241
nm and found to be proportional to the corresponding
concentrations of repaglinide.
For HPLC method, the standard solutions were
prepared by dilution of aliquots of the standard stock
solution with mobile phase to reach the linearity range
˜ȱśȬśŖȱΐȦ–•ȱ˜ȱ›Ž™Š•’—’Žǯȱ Ž—¢ȱ–’Œ›˜•’›Žȱ˜ȱ‘Žȱ
each standard solution was injected to HPLC system.
‘Žȱ™ŽŠ”ȱŠ›ŽŠœȱ Ž›Žȱ™•˜ĴŽȱŠŠ’—œȱ‘ŽȱŒ˜››Žœ™˜—’—ȱ
Preparation of sample solution
To determine the content of repaglinide in conventional
Š‹•Žœȱǻ•Š‹Ž•ȱŒ•Š’–DZȱŘȱ–ȱ›Ž™Š•’—’Žȱ™Ž›ȱŠ‹•ŽǼǰ 20
tablets were weighed, their mean weight determined
Š—ȱ ꗎ•¢ȱ ™˜ Ž›Žǯȱ ȱ ™˜›’˜—ȱ ˜ȱ Š‹•Žȱ ™˜ Ž›ȱ
equivalent to 10 mg of repaglinide was accurately
 Ž’‘ŽȱŠ—ȱ’œœ˜•ŸŽȱ’—ȱřŖȱ–•ȱ–Ž‘Š—˜•ȱ’—ȱŗŖŖȱ–•ȱ
volumetric flask. The contents of the flask were
sonicated for 15 min to dissolve repaglinide, volume
was made up to the mark with same diluent and the
›Žœž•’—ȱ–’¡ž›Žȱ ŠœȱꕝŽ›Žǯȱ—ȱŠ•’šž˜ȱ™˜›’˜—ȱ˜ȱ
˜‹Š’—Žȱꕝ›ŠŽȱ Šœȱ’•žŽȱ˜ȱŗŖȱ–•ȱ ’‘ȱ–Ž‘Š—˜•ȱȱ
Œ‘›˜–Š˜›Š™‘’ŒȱŠ—Š•¢œ’œȱ˜ȱŽȱꗊ•ȱŒ˜—ŒŽ—›Š’˜—ȱ
within the linearity range.
Method validation
 ‘ Ž ȱ ˜ ™  ’ – ’ £ Ž  ȱ œ ™ Ž Œ  › ˜ ™ ‘ ˜  ˜ – Ž  › ’ Œ ȱ Š — ȱ
chromatographic methods were completely validated
according to the procedure described in ICH guidelines
ŘȱǻŗǼȱ˜›ȱŸŠ•’Š’˜—ȱ˜ȱŠ—Š•¢’ŒŠ•ȱ–Ž‘˜œǯ
Linearity
’—ŽŠ›’¢ȱ  Šœȱ œž’Žȱ ‹¢ȱ Š—Š•¢£’—ȱ œ’¡ȱ œŠ—Š›ȱ
œ˜•ž’˜—œȱ ǻnȱ ƽȱ řǼȱ Œ˜ŸŽ›’—ȱ ‘Žȱ ›Š—Žȱ ˜ȱ śȬřŖȱ ΐȦ–•ȱ
Š—ȱ śȬśŖȱ ΐȦ–•ȱ ˜›ȱ ȱ œ™ŽŒ›˜™‘˜˜–Ž›’Œȱ Š—ȱ
HPLC, respectively. Standard solutions containing
 Dhole, et al.: Spectrophotometric and HPLC method for repaglinide

ŗŖŖȱΐȱȦ–•ȱ˜ȱ›Ž™Š•’—’Žȱ’—ȱœ˜•ŸŽ—ȱ Ž›Žȱ™›Ž™Š›Žȱ to evaluate the presence of possible interfering bands

in triplicate. Aliquots of these solutions were diluted at 241 nm.
˜ȱ œ’¡ȱ ’쎛Ž—ȱ Œ˜—ŒŽ—›Š’˜—œǰȱ Œ˜››Žœ™˜—’—ȱ ˜ȱ
˜ȱ śȬřŖȱ ΐȦ–•ȱ Š—ȱ śȬśŖȱ ΐȦ–•ȱ ˜ȱ ›Ž™Š•’—’Žȱ ˜›ȱ Ruggedness
UV spectrophotometric and HPLC, respectively. Ruggedness of the proposed method was determined
Calibration curves with concentration verses by analysis of sample solution prepared by proposed
Š‹œ˜›‹Š—ŒŽȱ ˜›ȱ ™ŽŠ”ȱ  Šœȱ ™•˜ĴŽȱ ˜›ȱ ŽŠŒ‘ȱ –Ž‘˜ȱ –Ž‘˜œȱ‹Ž ŽŽ—ȱ’쎛Ž—ȱ’–Žȱ’—Ž›ŸŠ•œǰȱŠ¢œȱŠ—ȱ
and the obtained data were subjected to regression Š—Š•¢œœǯȱ‘ŽȱƖȱǯǯǯȱ ŠœȱŽŽ›–’—Žǯ
analysis using the least squares method.
RESULTS AND DISCUSSION
Precision
Ž™ŽŠŠ‹’•’¢ȱ  Šœȱ ˜‹Š’—Žȱ ‹¢ȱ Š—Š•¢£’—ȱ œŠ–™•Žȱ
Method development and optimization
œ˜•ž’˜—ȱœ’¡ȱ’–ŽœǰȱŠȱŗŖŖƖȱ˜ȱŽœȱŒ˜—ŒŽ—›Š’˜—ȱ ’‘’—ȱ
Repaglinide was completely soluble in methanol
the same day using both methods. Similary, the intra
and hence methanol was selected as the solvent for
Š—ȱ’—Ž›ȱŠ¢ȱ™›ŽŒ’œ’˜—ȱ ŠœȱŽŸŠ•žŠŽȱ‹¢ȱŠ—Š•¢£’—ȱ
repaglinide to obtained UV spectrum in the range
Š‹•ŽȱœŠ–™•Žȱ˜—ȱ‘ŽȱœŠ–ŽȱŠ¢ȱŠ—ȱ˜—ȱ’쎛Ž—ȱŠ¢œȱ
˜ȱŘŖŖȬŚŖŖȱ—–ȱǽ’ž›ŽȱŘǾǯȱЎ›ȱ‘ŽȱŽŸŠ•žŠ’˜—ȱ˜ȱ‘Žȱ
Šȱ ’쎛Ž—ȱ ’–Žȱ ’—Ž›ŸŠ•ǰȱ ›Žœ™ŽŒ’ŸŽ•¢ǯȱ Ž™Š•’—’Žȱ
spectrum, the wavelength of 241 nm was selected for
Œ˜—Ž—œȱŠ—ȱ‘Žȱ›Ž•Š’ŸŽȱœŠ—Š›ȱŽŸ’Š’˜—ȱǻǯǯǯǼȱ
measurement, due to the adequate molar absorptivity
value were calculated.
of repaglinide in this region.
Accuracy
‘Žȱ Œ‘›˜–Š˜›Š™‘’Œȱ –Ž‘˜ȱ  Šœȱ ˜™’–’£Žȱ ‹¢ȱ
To check the accuracy of the developed methods
Œ‘Š—’—ȱ‘ŽȱŒ˜–™˜œ’’˜—ȱ˜ȱ–˜‹’•Žȱ™‘ŠœŽǰȱ̘ ȱ›ŠŽȱ
and to study interference of formulation additives,
and column. Finally development was carried out
analytical recovery experiments was carried out by
using C18 column and a mobile phase composed of
the standard addition method. Repaglinide reference
–Ž‘Š—˜•ȱŠ—ȱ ŠŽ›ȱǻŞŖDZŘŖȱŸȦŸǰȱ™
ȱŠ“žœŽȱ˜ȱřǯśȱ ’‘ȱ
standard solution was added to tablet samples at three
˜›‘˜™‘˜œ™‘˜›’ŒȱŠŒ’ǼȱŠȱŠȱ̘ ȱ›ŠŽȱ˜ȱŗǯŖȱ–•Ȧȹ–’—ǯȱ
’쎛Ž—ȱŒ˜—ŒŽ—›Š’˜—œȱ•ŽŸŽ•ǯȱȱŽŠŒ‘ȱ•ŽŸŽ•ǰȱœŠ–™•Žœȱ
The eluent was monitor at 241 nm. An adequate peak
were prepared in triplicate and the mean percentage
œ¢––Ž›¢ȱ ǻŠ’•’—ȱ ŠŒ˜›DZȱ ŗǯŘŘǼȱ Š—ȱ œ‘˜›ȱ ›ž—ȱ ’–Žȱ
›ŽŒ˜ŸŽ›¢ȱŠ—ȱǯǯǯȱŸŠ•žŽȱ Ž›ŽȱŽŽ›–’—Žȱ˜›ȱ‹˜‘ȱ
was achieved as demonstrated in the chromatogram
methods.
ǽ’ž›Žȱ řǾǯȱ ‘Žȱ œ¢œŽ–ȱ œž’Š‹’•’¢ȱ ™Š›Š–ŽŽ›œȱ Š›Žȱ
shown in Table 1.
Detection and quantitation limits
Series of diluted standard solutions were prepared
Linearity
Š—ȱŠ—Š•¢£Žȱ‹¢ȱ‹˜‘ȱ–Ž‘˜œǯȱ‘Žȱ•’–’ȱ˜ȱŽŽŒ’˜—ȱ
A linear relationship was found between the
ǻǼȱŠ—ȱ•’–’ȱ˜ȱšžŠ—’Š˜—ȱǻǼȱ Ž›ŽȱœŽ™Š›ŠŽ•¢ȱ
concentration and the response of both UV and HPLC
determined based on standard deviation of the
method. The regression analysis data are presented
y-intercept and the slope of the calibration curve by
’—ȱ Š‹•Žȱ Řǯȱ ‘Žȱ ›Ž›Žœœ’˜—ȱ Œ˜ŽĜŒ’Ž—œȱ ǻ›2Ǽȱ ˜‹Š’—Žȱ
žœ’—ȱ‘ŽȱŽšžŠ’˜—œȱǻŗǼȱŠ—ȱǻŘǼǰȱ›Žœ™ŽŒ’ŸŽ•¢ǯ

33δ ȱ ǻŗǼ
LOD =
S

33 δ
LOD = ȱ ǻŘǼ
S

‘Ž›Žǰȱ ΈDZȱ œŠ—Š›ȱ ˜ȱ ¢Ȭ’—Ž›ŒŽ™ȱ Š—ȱ DZȱ œ•˜™Žȱ ˜ȱ

calibration curve.

5peciſcity
A sample solution of tablet was prepared in the
test concentration range and injected into the
chromatograph, to evaluate possible interfering peaks.
For spectrophotometric analysis the UV spectrum of
this solution was recorded in the range of 200-400 nm Figure 2: UV spectra of repaglinide in methanol

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 Dhole, et al.: Spectrophotometric and HPLC method for repaglinide
 Šœȱ ‘’‘Ž›ȱ ‘Š—ȱ Ŗǯşşşȱ  ‘’Œ‘ȱ ŠĴŽœȱ ‘Žȱ •’—ŽŠ›’¢ȱ ˜ȱ
the methods.
Precision
The precision data obtained for the evaluated
methods are demonstrated in Table 2. Both methods
™›ŽœŽ—ŽȱǯǯǯȱŸŠ•žŽœȱ•˜ Ž›ȱ‘Š—ȱŘǯŖƖȱŠœœž›’—ȱŠȱ
good precision but HPLC method highly precise as
compared to UV method.
Accuracy
Accuracy was investigated by means of recovery
studies using the developed methods. Both
spectrophotometric and chromatographic methods
Ž¡‘’‹’Žȱ –ŽŠ—ȱ ›ŽŒ˜ŸŽ›’Žœȱ ǻn ȱ ƽȱ şǼȱ Œ•˜œŽȱ ˜ȱ ŗŖŖƖȱ
ǽŠ‹•ŽœȱřȱŠ—ȱŚǾǰȱŽ–˜—œ›Š’—ȱŠ—ȱŠŽšžŠŽȱŠŒŒž›ŠŒ¢ǯ
Ruggedness
‘ŽȱƖȱǯǯǯȱŸŠ•žŽœȱ›Ž™˜›Žȱ Ž›Žȱ˜ž—ȱ˜ȱ‹Žȱ•Žœœȱ
‘Š—ȱŘƖȱœ‘˜ Žȱ›žŽ—Žœœȱ˜ȱ‘Žȱspectrophotometric
and HPLC methods. The results of ruggedness were
™›ŽœŽ—Žȱ’—ȱŠ‹•Žȱřǯ
5peciſcity
The chromatogram obtained with the tablet sample
solution with excipients shows no interfering peaks in
Analysis of tablets
The quantitative results using UV and HPLC methods
Š›Žȱœ‘˜ —ȱ’—ȱŠ‹•Žȱśǯȱ‘Ž›Žȱ Šœȱ—˜ȱœŠ’œ’ŒŠ••¢ȱœ’—’ęŒŠ—ȱ
difference between the mean values, although UV
–Ž‘˜ȱœ‘˜ Žȱœ•’‘•¢ȱ‘’‘Ž›ȱƖȱǯǯǯȱŸŠ•žŽȱŒ˜–™Š›Žȱ
with HPLC method. Therefore, both analytical methods,
were found to be accurate, precise and could be used for
routine quality control analysis of repaglinide.
CONCLUSIONS
The advantage of UV method over HPLC method
is that the proposed UV method does not require
the elaborate treatment and procedures usually
associated with chromatographic method. It is
less time consuming and economical. A statistical
comparison of the quantitative determination of
repaglinide shows that HPLC method as more
accurate and precise than UV method. The results
indicate HPLC and UV spectrotometry methods are
adequate methods to quantify repaglinide in pure
form and its dosage form. There was no interference
Table 1: Result from system suitability study
Parameters* Repaglinide
the retention time of repaglinide. For the UV method, 5HWHQWLRQWLPHPLQ 
no interfering absorption band was found at 241 nm, 7DLOLQJIDFWRU 1.22
7KHRUHWLFDOSODWHV1 
in the spectrum of the tablet sample solution.
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Š—ȱ řǯŚŞȱ ΐȦ–ǰȱ Ŗǯŝřȱ ΐȦ–ȱ Š—ȱ ŘǯŘŗȱ ΐȦ–ȱ ˜›ȱ
spectrophotometric and HPLC methods, respectively. Table 2: Validation parameters of evaluated
methods
Parameters UV HPLC
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Figure 3: Chromatogram of standard solution of repaglinide 6SHFL¿FLW\ 6SHFL¿F 6SHFL¿F

Table 3: Recovery study results of repaglinide for UV method

Level of % recovery Amount spiked (μg/ml) Amount recovered (μg/ml) % Mean recovery % R.S.D.* (n = 3)
    
100 20  “ 
120 24 24.12 “ 
ȗǤǤγ‡Žƒ–‹˜‡•–ƒ†ƒ”††‡˜‹ƒ–‹‘

Pharmaceutical Methods | July-December 2012 | Vol 3 | Issue 2

71
 Dhole, et al.: Spectrophotometric and HPLC method for repaglinide

Table 4: Recovery study results of repaglinide for HPLC method

Level of % recovery Amount spiked (μg/ml) Amount recovered (μg/ml) %Mean recovery % R.S.D.* (n = 3)
    
100 20   
120 24   
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’—‹ž›‘DZȱ‘ž›Œ‘’••ȱ’Ÿ’—œ˜—ŽDzȱŘŖŖřǯȱ™ǯȱřŗŖǯ
Table 5: Statistical comparison of results 2. Budavari S, editor. The Merck Index, an encyclopedia of chemicals,
obtained by the proposed methods for the ›žœȱŠ—ȱ‹’˜•˜’ŒŠ•œǯȱŗřthȱŽǯȱ‘’Ž‘˜žœŽȱŠ’˜—ǰȱDZȱŽ›Œ”ȱŠ—ȱ˜ȱ
analysis of repaglinide in tablets —ŒDzȱŘŖŖŗǯȱ™ǯȱŝşŖǯ
Drug Label claim Parameter UV HPLC řǯȱ Ž¢—˜•œȱ ǯȱ Š›’—Š•Žǰȱ ‘Žȱ Œ˜–™•ŽŽȱ ›žȱ ›ŽŽ›Ž—ŒŽǯȱ řřrd ed.
mg/tablet (n = 6) method ˜—˜—DZȱ‘Š›–ŠŒŽž’ŒŠ•ȱ›ŽœœDzȱŘŖŖŘǯȱ™ǯȱřřŚǯ
5HSDJOLQLGH 2.0 0HDQ   4. Goyal A, Singhvi I. Visible spectrophotometric methods for estimation
of repaglinide in tablet formulation. —’Š—ȱȱ‘Š›–ȱŒ’ȱŘŖŖŜDzśDZŝşȬŞŖǯ
$PRXQWIRXQGPJ  
5. Sharma S, Sharma MC. Simultaneous determination of repaglinide
6'   in pharmaceutical dosage form using indigo carmine. Amer Eurasian
56'   ȱŒ’ȱŽœȱŘŖŗŗDzŜDZŚŝȬśŗǯ
ȗǤǤǣ–ƒ†ƒ”†‡˜‹ƒ–‹‘ǢǤǤǤǣ‡Žƒ–‹˜‡–ƒ†ƒ”†‡˜‹ƒ–‹‘ Ŝǯȱ ŠŽ•ȱ ǰȱ ž‘Š’Šȱ ǰȱ ŠŽ•ȱ 
ǯȱ ’–ž•Š—Ž˜žœȱ œ™ŽŒ›˜™‘˜˜–Ž›’Œȱ
estimation of metformin and repaglinide in a synthetic mixture.
—’Š—ȱȱ‘Š›–ȱŒ’ȱŘŖŖŝDzŜşDZŞŚŚȬśŘǯ
by excipients in the tablets and the mobile phase is ŝǯȱ Šžœ‘Š•ȱǰȱŠ’—ȱǰȱ’ Š›¢ȱ ǯȱŽŸŽ•˜™–Ž—ȱ˜ȱœ™ŽŒ›˜Ěž˜›’–Ž›’Œȱ
easy to prepare. Since these methods are simple, and HPLC methods for in vitroȱ Š—Š•¢œ’œȱ ˜ȱ ›Ž™Š•’—’Žǯȱ —’Š—ȱ ȱ
œ™ŽŒ’ęŒǰȱ ›Š™’ǰȱ ™›ŽŒ’œŽȱ Š—ȱ ŠŒŒž›ŠŽǰȱ ‘Ž¢ȱ –Š¢ȱ ‹Žȱ ‘Š›–ȱŒ’ȱŘŖŗŖDzŝŘDZŘŚŖȬŘǯ
8. Š—‘’–Š‘’ȱ ǰȱ Ž—žȱ  ǯȱ ŽŽ›–’—Š’˜—ȱ ˜ȱ ›Ž™Š•’—’Žȱ ’—ȱ
successfully and conveniently adopted for routine
pharmaceutical formulation by HPLC with UV detection. Anal Sci
quality control analysis of repaglinide in bulk and ŘŖŖřDzŗşDZŗŜŝśȬŞŘǯ
pharmaceutical dosage form. şǯȱ ›Š–ŽŽ•ŠȱǰȱŠ•Šȱǰȱ›Œ‘Š—Šȱǰȱ’ŸŠȱǰȱ›ž—ŠȱǯȱŽŽ›–’—Š’˜—ȱ
˜ȱ ›Ž™Š•’—’Žȱ ’—ȱ ™‘Š›–ŠŒŽž’ŒŠ•ȱ ˜›–ž•Š’˜—ȱ ‹¢ȱ
ǯȱ  Applied
Sci Res ŘŖŖşDzśDZŗśŖŖȬŚǯ
ACKNOWLEDGMENTS ŗŖǯȱ ž£’•Š Š’ȱ ǰȱ Š‘Š‹ȱ ǰȱ –›Š—ȱ ǰȱ œ–Š’•ȱ ǰȱ Š—Šȱ 
ǯȱ Ž‘˜ȱ
development and validation of repaglinide in human plasma by

ȱ Š—ȱ ’œȱ Š™™•’ŒŠ’˜—ȱ ’—ȱ ™‘Š›–ŠŒ˜”’—Ž’Œȱ œž’Žœǯȱ ȱ ‘Š›–ȱ
Authors are thankful to the Manager, USV Lab. Pvt. Ltd.,
’˜–Žȱ—Š•ȱŘŖŖŝDzŚřDZŗŞřŗȬśǯ
ž–‹Š’ǰȱ —’Šȱ ˜›ȱ ™›˜Ÿ’’—ȱ ‘Žȱ ’Ğȱ œŠ–™•Žœȱ Š—ȱ Š•œ˜ȱ 11. 
ȱ ž’Ž•’—Žœǰȱ Řǰȱ Š•’Š’˜—ȱ ˜ȱ —Š•¢’ŒŠ•ȱ ›˜ŒŽž›ŽœDZȱ
‘Š—”ž•ȱ ˜ȱ ›ǯȱ ǯȱ ǯȱ ‘žœŠ›’ǰȱ ›’—Œ’™Š•ǰȱ ‘Š›Šȱ Š Š›ȱ Ž‘˜˜•˜¢ǯȱ—Ž›—Š’˜—Š•ȱ˜—Ž›Ž—ŒŽȱ˜—ȱ
Š›–˜—’£Š’˜—ǯȱ Ž—ŽŸŠDZȱ
College of Pharmacy, Nagpur for providing experimental  ’£Ž›•Š—DzȱŗşşŜǯ
facilities for this work.
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