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New Development Method for Determination of Cefuroxime Axetil (CUA) and Cefprozil (CZ) In Pharmaceutical Drugs By RP-HPLC
M A Alfeen 1* and Y Yildiz2
1Department of Chemistry, Faculty of Second Science, Al-Baath University, Homs, Syria
2Science Department, Centenary University, Hackettstown New Jersey, USA
Submission: July 7, 2019; Published: July 30, 2019
*Corresponding author: M A Alfeen, Department of Chemistry, Faculty of Second Science, Al-Baath University, Homs, Syria
How to cite this article: M A Alfeen, Y Yildiz. New Development Method for Determination of Cefuroxime Axetil (CUA) and Cefprozil (CZ) In Pharmaceutical
Drugs By RP-HPLC. Organic & Medicinal Chem IJ. 2019; 8(4): 555744 DOI: 10.19080/OMCIJ.2019.08.555744
High performance liquid chromatography was one of the most important technologies used in drug control and pharmaceutical quality control. In this study, an analytical method was developed using chromatography method for determination of two Cephalosporin as like: Cefuroxime Axetil (CUA), and Cefprozil (CZ) in pharmaceutical Drug Formulations. Isocratic separation was performed on an Enable C18 column (125mm × 4.6mm i.d, 5.0μm, 10A˚) Using Triethylamine: Methanol: Acetonitrile: Ultra-Pure Water (0.1: 5: 25: 69.9 v/v/v/v %) as a mobile phase at flow rate of 1.5 mL\min. The PDA detection wavelength was set at 262nm. The linearity was observed over a concentration range of (0.01–50μg\mL) for RP-HPLC method (correlation coefficient=0.999). The developed method was validated according to ICH guidelines. The relative standard deviation values for the method precision studies were < 1%, and an accuracy was > 98%. The results were within the limits allowed by the US Pharmacopoeia and successfully for determination of Cefuroxime Axetil (CUA) and Cefprozil (CZ) in Tablets and dry syrup of the local pharmaceutical formulations.
Cefuroxime Axetil (CUA) is a second-generation of cephalosporin using as antibiotic for Drug Pharmaceutical: (6R, 7R)-3-(carbamoyloxymethyl)-7-[[(2Z)-2-(furan-2-yl)-2-methoxyim inoacetyl] amino]-8-oxo-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (Figure 1) . Cefprozil (CZ) is a second-generation cephalosporin using as antibiotic for Drug Pharmaceutical: (6R,7R)-7-[[(2R)-2-amino-2-(4-hydroxyphenyl) acetyl] amino]-8-oxo-3-[(E)-prop-1-enyl]-5-thia-1-azabicyclo [4.2.0] oct-2-ene-2-carboxylic acid (Figure 2) .
Most cephalosporins were used as broad-spectrum antibiotics, especially urinary and genital tract infections, and Gram-negative tests towards anaerobic or pathogenic bacteria were also important for the treatment of pharyngitis and MRI, since these two substances are of great local importance in Syria, as they were produced in multiple pharmaceutical forms. As a result of the great pressure on chemists to follow each production line on one, the conditions of method have been developed by researchers to save time and effort in quality control according to international and local standards. CUA and CZ have beenfound to reduce the corrosion of mild steel in hydrochloric acid
solution . According to literature surveys, there are different
analytical methods reported for the determination of CUA and
CZ. It includes UV-Vis spectroscopy [2-5], chemiluminescence
, HPLC [7-10]. Specificity and stability parameters for the
drug were assessed according to ICH .
Cefuroxime as Axetil (CUA) and Cefprozil (CZ) (purity of
all > 99.80%) was obtained as a gift sample from Parabolic
Pharmaceuticals Ltd., India. Analytical grade Methanol,
Acetonitrile, Triethylamine (Merck GmpH). The Ultra-Pure water
for HPLC was obtained by using TKA Water Purification System,
Germany. All Pharmaceutical Drug formulation was bought from
the local market (Table 1).
RP-HPLC System. An Enable C18 (125mm × 4.6mm, i.d
5μm, 10A˚) was used for separation. Triethylamine: Methanol:
Acetonitrile: Ultra-Pure Water (0.1: 5: 25: 69.9 v/v/v/v%) as a
mobile phase at flow rate of 1.5mL\min in isocratic mode. The
PDA detection was carried out at 262nm (Figure 3).
Standard stock solutions of CUA and CZ were prepared to
make 1000μg\mL solutions. Make Sample solutions as standard
by weigh of content Powder Tablets and Dry for oral suspension
equivalent to 100mg of CUA and CZ was accurately measured,
and transferred into two separate 100mL volumetric flasks,
containing 10mL of diluents mobile phase and ultrasonicated
for 15minutes; the volume was made up and mixed well.
Solutions were filtered by a 0.2μm filter to remove particulate
matter. Filtered solutions were properly diluted for analysis as
already described. The drug present in the sample solutions was
calculated by using the calibration curves. All the solutions were
stored at (2-8) ºC for the future use.
A ten-point (0.01, 0.02, 0.08, 1.0, 2.0, 8.0, 10, 20, 40, and
50μg\mL) calibration curves were prepared. The peak area for
HPLC method was obtained by injecting 20μl into the column.
Calibration curves were plotted by taking the peak area curve on
the y-axis and the concentration μg\mL on the x-axis.
The intraday and interlay precision study was carried out to
check the reproducibility of results. A concentration of 0.04, 0.4,
4.0 μg\mL and 12μg\mL of CUA and CZ (n=6) were analyzed to
find out relative standard deviation RSD% for RP-HPLC method.
To check the accuracy of the proposed method, recovery
studies were carried out at (1, 80, 100, and 120%) of the test
concentration. The recovery study was performed three times at
each level. The amount of CUA and CZ present in the sample was
calculated using calibration curves.
The robustness of RP-HPLC method was studied by
deliberately changing method parameters like flow rate of mobile
phase, detection wavelength, and organic phase compositions.
A series of system suitability parameters like retention time,
theoretical plates, and tailing factor were determined for each
changed condition according to ICH .
The LOD and LOQ were determined separately according to
the ICH guidelines. For HPLC method, concentrations providing
a signal-to-noise ratio 3:1 and 10:1 were considered as the LOD
and LOQ, respectively.
Optimization of mobile phase was carried out based on
tailing factor and theoretical plates obtained for CUA and CZ.
During the trial runs, the drug was tested with different mobile
phase compositions like (Triethylamine: Methanol: Acetonitrile:
Ultra-Pure Water v/v/v/v%) at various compositions (10:10:
20: 60 v/v/v/v%), (5: 15: 15: 65 v/v/v/v%), and (1: 5: 20: 74v/v/v/v%) and flow rates (1.0,1.5, and 2.0 mL\min). The mobile
phase consisting of Triethylamine: Methanol: Acetonitrile: Ultra-
Pure Water (0.1: 5: 25: 69.9 v\v\v\v%) at a flow rate of 1.5 mL\
min was selected which gave a sharp, symmetric peak for CUA,
CZ. The retention time for CUA, CZ was found to be (2.86, 4.18
min) respectively. The run time was 5min. The tailing factor
for CUA, CZ was found to be (1.12, 1.17). PDA detection was
carried out at 260 nm. The separation was carried out at room
temperature (Figure 4) respectively.
To evaluate the specificity, PDA detector was applied to find
out the peak purity of the chromatographic peaks obtained for the
stress-treated drug solution. Peak purity results are indicative for
finding out the peak homogeneity. The specificity of the RP-HPLC
method was determined by checking the interference of any of
the possible degradation products and Absorbances produced
during study of CUA, CZ. The study of the drug was carried outwith Methanol, Triethylamine, Ultra-Pure Water for discovering
the stability nature of the drug. The degraded samples were
prepared by taking suitable aliquots of the drug solution, and
then undertaking the respective stress testing procedures for
each solution. The chromatograms (Figure 5) obtained for the
blank and placebo show no interference due to solvent used
and presence of the commonly used excipients suggesting the
specificity of two methods.
The calibration curves were found to be linear over a
concentration range of (0.01–50 μg\mL) for method (correlation
coefficient 0.999 for all method). The method parameters and
regression data are shown in (Table1).
The HPLC method was found to be robust under deliberate
changes in the mobile phase flow rate (±0.1 mL\min), detection
wavelength (±5nm), and organic phase composition (±2%). For
the UV spectroscopic methods, changing the slit width shows no
significant effect on absorbance, indicating the robustness of the
developed methods. No significant changes were obtained in the
content of CUA, CZ during the solution stability studies by the
developed methods. The recoveries for the solution stability by
Method was found to be 99.89%, and 100.21%, respectively.
The proposed chromatographic method in this study was
characterized by high accuracy and excellent repetition through
the very low standard deviation values, good health through
the good return values compatible with the constitutional field,
in addition to the ease and speed of quality control without
obstruction of additives to the pharmaceutical, Using them
in the analysis of pharmaceuticals, and their application in
pharmaceutical control laboratories.
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