ArticlePDF Available

Validated Spectrophotometric Method for the Estimation of Ketorolac Tromithamine in Bulk and Tablets Using Ninhydrin: A Modified Approach

Authors:

Abstract and Figures

A new, accurate, precise ecofriendly and economical spectrophotometric method has been developed and validated for the determination of Ketorolac Tromithamine in bulk drugs and tablet dosage forms. The proposed method is based on the reaction of ninhydrin with primary amino group present in Ketorolac Tromithamine in the presence of Ammonium Molybdate which forms Ruhemann’s purple colour by heating at 90 ± 1°C for 15 min and absorbs maximally at about 570 nm. Beer-Lambert’s law is obeyed in the concentration range of 50-250μg/ml and is described by the regression equation Y = 0.002x – 0.186 with a regression coefficient (r2) = 0.998 (n = 6). The effects of variables such as temperature, heating time, concentration of colour producing reagent and stability of colour were investigated to optimize the procedure. For the proposed method, the value of molar absorptivity and Sandell’s sensitivity are 1.03434 x 103 L/mol/cm and 0.363995μg/cm2, respectively. The LOD and LOQ are found to be 15.369 μg/ml and 46.57μg/ml, respectively. The statistically validated results indicate that the proposed method has good sensitivity, accuracy, precision and stability. Along with its cost effect, it is eco friendly and used for routine quality control of Ketorolac Tromethamine in bulk and tablet dosage forms.
Content may be subject to copyright.
Asian J. Research Chem. 7(1): January 2014
19
ISSN 0974-4169
www.ajrconline.org
RESEARCH ARTICLE
Validated Spectrophotometric Method for the Estimation of Ketorolac
Tromithamine in Bulk and Tablets Using Ninhydrin: A Modified Approach
Vandana P. Patil*, Subhash J. Devdhe, Sachidanand S. Angadi, Suwarna H. Kale ,
Shital D. Phalke, Santosh D. Shelke and Rushikesh H. Patil
Yash Institute of Pharmacy, Department of Pharmaceutical Analysis,Aurangabad-431134, India
*Corresponding Author E-mail: vandana2609@gmail.com
ABSTRACT:
A new, accurate, precise ecofriendly and economical spectrophotometric method has been developed and validated for
the determination of Ketorolac Tromithamine in bulk drugs and tablet dosage forms. The proposed method is based on
the reaction of ninhydrin with primary amino group present in Ketorolac Tromithamine in the presence of Ammonium
Molybdate which forms Ruhemann’s purple colour by heating at 90 ± 1°C for 15 min and absorbs maximally at about
570 nm. Beer-Lambert’s law is obeyed in the concentration range of 50-250µg/ml and is described by the regression
equation Y = 0.002x – 0.186 with a regression coefficient (r
2
) = 0.998 (n = 6). The effects of variables such as
temperature, heating time, concentration of colour producing reagent and stability of colour were investigated to
optimize the procedure. For the proposed method, the value of molar absorptivity and Sandell’s sensitivity are 1.03434
x 10
3
L/mol/cm and 0.363995µg/cm
2
, respectively. The LOD and LOQ are found to be 15.369 µg/ml and 46.57µg/ml,
respectively. The statistically validated results indicate that the proposed method has good sensitivity, accuracy,
precision and stability. Along with its cost effect, it is eco friendly and used for routine quality control of Ketorolac
Tromethamine in bulk and tablet dosage forms.
KEY WORDS:
Ketorolac Tromithamine; Ninhydrin; Ammonium Molybdate; UV-visible Spectrophotometry.
1.0 INTRODUCTION
Ketorolac Tromithamine (KT) is a NSAID in the family of
Heterocyclic Acetic Acid derivative. Chemically, it is 5-
Benzoyl-2,3-Dihydro-1H-Pyrolizine-1-Carboxylic Acid, 2
Amino-2-(hydroxymethyl)-1,3-Propaneddiol (Figure1)
1
. KT
acts by inhibiting the bodily synthesis of prostaglandins. It
is used in short term management of moderate to severe
pain
2
.
N
O
HO
NH2
OH
OH
OH
O
.
Figure 1: Structure of Ketorolac Tromithamine (KT).
European Pharmacopoeia describes assay of Ketorolac
Tromithamine by reversed phase high performance liquid
chromatography in bulk and pharmaceutical formulations
3
.
KT has been studied and determined by relatively other
methods such as Spectrophotometry
4-6
, HPLC
7-12
and
HPTLC
13-16
.
Received on 10.08.2013 Modified on 20.10.2013
Accepted on 06.11.2013 © AJRC All right reserved
Asian J. Research Chem 7(1): January 2014; Page 19-24
Many researchers have dealt with the development of
methods that quantify Ketorolac Trimithamine (KT) in pure
form and in tablets. Visible spectrophotometry is the
technique of choice even today because of its inherent
simplicity, sensitivity, selectivity, accuracy, precision and
cost-effectiveness. The scientific references found in the
CAS and SCI database, relating to green analytical
chemistry or environmentally-friendly analytical methods
have been growing significantly in recent years
17-18
. The
recent development of new analytical methods with good
characteristics such as selectivity and sensitivity are not
only sufficient but also modern analytical methods need to
be green. Hence, the purpose of the present study was to
improve these methods with respect to two experimental
objectives: (i) decrease the heating time, (ii) avoid the use
of organic solvent. Both factors are of great significance in
reducing time and cost of analysis. In the present
investigation, solution of distilled water has been employed
to solubilize a water soluble drug, KT forms a purple
colored product with ninhydrin in the presence of saturated
solution of Ammonium Molybdate
and further
spectrophotometric estimation was carried out maximum at
about 570 nm without employing any organic solvents.
These modifications resulted in increased linear range (50-
250µg/ml), molar absorptivity (ε = 1.034 x 10
3
L/mol/cm)
Asian J. Research Chem. 7(1): January 2014
20
and sensitivity (0.363901µg/cm
2
) compared too many
existing spectrophotometric methods.
2.0 EXPERIMENTAL
2.1 Apparatus
A Shimadzu 1800 double beam UV-VIS spectrophotometer
provided with 1 cm matched quartz cell was used for
absorbance measurements.
2.2 Materials and Reagents
Ketorolac Tromithamine was obtained as gift sample from
Wockhardt Pharmaceuticals Pvt. Ltd. (Aurangabad, India).
All other reagents used were of analytical grade.
2.3 Solubility Studies
Ketorolac Tromithamine is water soluble and the proposed
method was carried out in distilled water.
2.4 Preparation of Citrate buffer
10.5gm Citric acid was dissolved in 500ml of 1M NaOH.
2.5 Preparation of 0.2M Ninhydrin Solution
0.44gm mg of Ninhydrin was dissolved in 25 ml of Citrate
buffer.
2.6 Preparation of 0.2 M Ammonium Molybdate
Approximately 6.1gm was taken in a beaker containing 25
ml of citrate buffer and stirred with a magnetic stirrer for
twenty minutes. The solution was decanted and filtered
using quantitative filter paper.
2.7 Preparation of Standard solution of KT
100 mg pure Ketorolac Tromithamine was transferred to
1000 ml volumetric flask and diluted upto the mark with
distilled water to get a concentration of 100µg/ml solution.
2.8 Optimized Procedure
In a 10 ml volumetric flask, add 0.5 ml standard stock
solution (100µg/ml), 1.0 ml ninhydrin and 1.0 ml
ammonium molybdate solution and diluted upto the mark
with citrate buffer (10µg/ml). It was heated in boiling water
bath at 90 ± 1°C in 15 min and cooled to room temperature.
Scan the spectrum and absorbance was measured λmax at
570 nm versus the reagent blank (Figure 2).
Figure 2: Spectrum of Ketorolac Tromithamine (KT)
3.0 METHOD VALIDATION:
3.1 Stability
In order to demonstrate the stability of the Ninhydrin-
Ketorolac Tromithamine complex in presence of
Ammonium molybdate solution was analyzed over a period
of 12 hrs at room temperature. During analysis, the complex
was found to be stable over a period of 2 hr at room
temperature as shown in Figure 3.
Figure 3: Effect of time (min) on absorbance of KT- Ninhydrin
complex.
3.2. Linearity
From the Standard Stock solution (100µg/ml), different
aliquots (0.5, 1.0, 1.5, 2.0 and 2.5) were taken in a series of
10 ml volumetric flasks and 1.0 ml ninhydrin was added to it
followed by 1.0 ml Ammonium molybdate solution and
volume made up with Citrate buffer to get concentration 50-
250µg/ml. All flasks were heated for 15 min in boiling water
bath and cooled to room temperature and measure the
absorbance at 570 nm. Five replicates of analytes were
measured and record the absorbance. Plot a graph
concentration versus absorbance, a linear correlation was
found which obeys Beer Lambert’s Law in the concentration
range of 50-250 µg/ml (Figure 4). Regression analysis of
Beer’s law data using the method of least squares was made
to evaluate the slope (b), intercept (a) and the correlation
coefficient (r
2
).
Figure 4: Calibration curve of Ketorolac Tromithamine (KT).
Asian J. Research Chem. 7(1): January 2014
21
3.3 Limit of Detection (LOD) and Limit of
Quantification (LOQ)
The detection limit of an individual analytical procedure is
the lowest amount of analyte in a sample that can be
detected but not necessarily quantitated as an exact value.
The quantitation limit of an individual analytical procedure
is the lowest amount of analyte in a sample that can be
quantitatively determined with suitable precision and
accuracy. The quantitation limit is a parameter of
quantitative assays for low levels of compounds in sample
matrices and is used particularly for the determination of
impurities and/or degradation products. The valve of LOD
and LOQ are determined and given in Table 1.
Table 1: Optical characteristics data and validation parameters of
KT.
Parameter Analytical data
Linearity Range (µg/ml) 5-25
λ max (nm) 570
Molar extinction coefficient 1.034x10
3
Sandell’s sensitivity 0.363901
Slope 0.7 x 10
-
2
Intercept -205
Standard deviation about regression (Sy) ±0.241077
Standard deviation of Slope (Sb) ±0.001525
Standard deviation of intercept (Sa) ±0.252844
Correlation co-efficient (r
2
) 0.997
Limit of detection (LOD, µg/ml) 15.36995
Limit of quantification (LOQ, µg/ml) 46.5756
3.4. Precision
To determine precision, 7 days measurement (intra-days
and interday) were computed with relative standard
deviation (RSD %) for replicate samples (n = 5) using
concentration 50, 100and 150µg/ml both the intraday and
interday samples were calibrated with standard curve
concurrently prepared in the same day of analysis.
3.4.1. Intraday Precision
Intraday precision of test method is demonstrated by three
samples of the same batch (same concentration) at initial,
24 and 48 hrs (Table 2).
3.4.2. Interday Precision
Interday precision of test method is demonstrated by three
samples of the same batch (same concentration) on three
successive days (Table 2).
3.5. Accuracy
To determine the accuracy of the proposed method,
recovery study was carried out by adding different amount
(80%, 100%. 120%) of bulk sample of Ketorolac
Tromithamine within the linearity range and results
obtained are compiled in Table 3 and show good accuracy
for the method.
3.6 Assay
Assay of tablet dosage form was carried by same procedure
as mentioned in methodology to equivalent weight of
Ketorolac Tromithamine by proposed spectrophotometric
method. The percent purity was found out using regression
analysis (Table 4).
4.0 RESULTS AND DISCUSSION:
Preliminary studies were carried out to establish the
optimum conditions for assay of the Ketorolac
Tromithamine.
4.1 Effect of temperature
The effect of temperature on the complexation reaction at
80
0
, 90
0
and 97 ± 1°C were examined (Table 5). It was
observed that ninhydrin–AM complex in saturated
Ammonium Molybdate required 97 ± 1°C for obtaining
maximum and stable absorbance and remained constant for
about a further 2 hrs.
4.2 Effect of the reaction heating time
The effect of time on the complexation reactions at room
temperature was examined. It was observed that ninhydrin–
KT charge transfer complex in saturated sodium
bicarbonate required 15 min for obtaining maximum and
stable absorbance as shown in Figure 5.
Table 2: Evaluation of intra-day and inter-day accuracy and precision
KT taken
(µg/ml)
Intraday Accuracy and precision Interday Accuracy and precision
KT found(µg/ml) RE % RSD % KT found(µg/ml) RE % RSD %
100 89.29 89.29% 1.39 85.74 85.74 1.47
150 143.75 95.83% 0.87 139.62 93.08 0.89
200 191.24 95.60% 0.65 188.54 94.27 0.66
Table 3: Recovery Data
Level Amount of KT added (µg) Amount of KT found (µg) % Recovery % RSD
80 % 80 82.40 103 0.68
100 % 100 102.67 102.67 0.82
120 % 120 111.14 92.61 1.21
*An average value ± relative standard deviation of 5 observations.
Table 4: Assay Results of Tablet Dosage Form.
Formulation Actual amount of KT (µg) Amount found of KT(µg) % of KT Found
Tablet 10 10.13 101.3
Asian J. Research Chem. 7(1): January 2014
22
Table 5: Effect of temperature on absorbance.
Temperature (
0
C) Absorbance
80 ± 1 0.042
90 ± 1 0.904
97 ± 1 1.644
Figure 5: Effect of the reaction heating time on absorbance.
4.3 Effect of solvent
Distilled water was a solvent of choice for ninhydrin–KT
system because intense and stable colour was obtained only
after 10 min of reagent mixing.
4.4 Effect of reagents concentration
The optimum conditions for the method was established by
varying the concentration of reagent at a time and keeping
the fixed drug concentration and observing the effect
produced on the coloured species. To establish the optimum
experimental condition for ninhydrin–KT charge transfer
complex, the drug (100 µg /ml) was allowed to react with
varying volumes of 0.5%ninhydrin.The maximum
absorbance was obtained with 1.0 ml of the reagent (Figure
6); further addition caused no significant change in
absorbance. Therefore a volume of 1.0 ml was used as an
optimum value.
Figure 6: Effect of volume of 0.5% ninhydrin on the absorbance of
reaction product
O
O
O H
O H
2
N i n h y d r i n
O
O
O H
H O
H y d r i n d a n t in
O
O
R u h e m a n n ' s P u r p l e
A m m o n i u m M o l y b d a t e
+
+
1 5 M i n u t e s
9 0 ± 1 ° C
N
O
O
O
O
N
O
H O
N H
2
O H
O H
O H
O
.
N
O
H O
O H
O H
O H
O H
O
.
K e t o r o l a c T r o m i t h a m i n e
Scheme I: Formation of Ruhemann’s purple complex between KT and ninhydrin.
Asian J. Research Chem. 7(1): January 2014
23
Table 6: Comparison of Proposed and Published SpectrophotometricMethods
19-20
.
It was reported that in alkaline medium, ninhydrin is
reduced to an intermediate, hydrindantin and converted to
diketohydrindylidene diketohydrindamine which is
commonly referred as ‘Ruhemann’s purple’ The primary
amino group of Ketorolac Tromithamine reacted with
ninhydrin in presence of saturated Ammonium Molybdate
solution (alkaline medium) to give diketohydrindylidene
diketohydrindamine of Ketorolac Tromithamine reacts with
ninhydrin via oxidation deamination of the primary amino
group followed by the condensation of the reduced
ninhydrin to form the colored Ruhemann’s purple without
employing any organic solvent., which absorbs a maximum
at 570 nm as shown in Figure 2. The proposed reaction
between of Ketorolac Tromithamine and ninhydrin is
shown in the Scheme I.
To optimize the reaction conditions, different parameters
have been investigated such as temperature, heating time,
reagent concentration, and color stability. Reaction between
ninhydrin and of Ketorolac Tromithamine did not give any
colored product in the absence of Ammonium Molybdate,
not even after prolonged heating. It was observed that
complete colour development was attained at 90± 1°C. The
optimum reaction time was determined by heating the
reaction mixture on a water bath at 90± 1°C. It was noted
that complete colour development was attained in fifteen
minutes. The effect of ninhydrin concentration on the
colour development was investigated. 1 ml of 0.5%
ninhydrin reagent produced maximum colour intensity.
Interestingly, reaction was found to be specific in
Ammonium Molybdate
medium. The comparison between
the proposed and published method was shown in Table 6.
5.0 CONCLUSIONS:
By the comparison of proposed and published methods as
shown in Table 6, it can be concluded that
spectrophotometric method has found to be new for the
determination of Ketorolac tromethamine. In comparison
with the existing visible spectrophotometric methods for the
quantification of Ketorolac Tromethamine, the present
modified method can be considered green as it
demonstrates that visible spectrophotometry can be utilized
without the usage of organic solvent. Overall the proposed,
spectrophotometric method is new, accurate, precise,
economical and ecofriendly. Thus it can be suitable for
quality control of Ketorolac Tromethamine in bulk, fixed-
dose combination tablets.
6.0 ACKNOWLEDGEMENT:
The authors are gratefully acknowledging the receipt of
pure Ketorolac Tromethamine as gift sample from
Wockhardt Pharmaceuticals Pvt. Ltd.(Aurangabad, India).
Authors express their gratitude to Yash Institute of
Pharmacy, Aurangabad for providing the instrumental and
chemicals facility.
7.0. REFERENCES:
1. USP Vol. 1.United States Pharmacopoeial Convention;
2003:1049.
2. Norvasc, Online Drug Description and Clinical Pharmacology of
ketorolac, The Internet Drug Index, Rx List Inc., 2008.
Website:http://www.rxlist.com/cgi/generic/keto2.htm.
3. Tripathi K.D., “Essential of Medical Pharmacology”,5
th
Edn.,
Jaypee Brothers Medical Publisher; New Delhi;
4. KumaraSwamy G., Kumar JMR., Sheshagirirao J.V.L.N.;
Simultaneous Estimation of Febuxostat and Ketorolac in
Pharmaceutical Formulations by Spectroscopic Method;
International Journal of ChemTech; 2012; 4:847-850.
5. Fegade J.D., Mehta H.P., Choudhari R.Y., Patil V.R.;
Simultaneous Spectrophotometric Estimation of Ofloxacin and
Ketorolac Tromethamine in Opthalmic Dosage Form;
International Journal of ChemTech; 2009;1;189-194.
6. Dewani A.P., Bakal R.L., Dr.Shiradkar M.R., Dr.Chandewar
A.V.;Absorpion Ratio Method for the Estimation of
Moxifloxacin HCL and Ketorolac Tromethamine in their
Combined Dosage Form by UV-Visible Spectroscopy;
International Journal of Pharmaceutical Research and
Development; 2011; 3(7);21-26.
7. Sunil G., Jambulingam M., Thangadurai S. A., Kamalkannan D.,
Sundaraganapathy R., Jothimanivannam C., Development and
Validation of Ketorolac Tromithamine in Eye Drop Formulation
by RP-HPLC Method; Arabian Journal of Chemistry; 2013.
8. Irene Tsina, Yuen Ling Tam, Aeelin Boyd, Cynthia Rocha, Ian
Massey, Thomas Tarnowski; An Indirect and Direct HPLC
Method for the determination of the Enantiomers of Ketorolac in
Plasma; Journal of Pharmaceutical and Biomedical Analysis;
1996; 15(3); 403-417.
9. Yang Guo Ping, HE Bin,LIU Shi Kun,et al; Determination of
Content of Ketorolac Tromethamine in Injection by HPLC;
Chinese Journal of Hospital Pharmacy; 2012.
10. Tsvetkova Boyka G., Pencheva, Ivanka P., Peikov, Plamen T.;
HPLC Determination of Ketorolac Tromethamine in Tablet
Dosage Forms; Der Pharmacia Sinica; 2012; 3(4); 400.
Parameter Proposed Method Published Methods
I II III
Solvent Distilled Water Methanol Phosphate Buffer Methanol
Reagents
Citrate buffer pH
5.5
2,3-dichloro-5,6-dicyano-p-
benzoquinone (DDQ)
Methylene Blue 2,3-dichloro-5,6-dicyano-p-
benzoquinone (DDQ)
Ammonium
molybdate
2,4-dichloro-6-nitrophenol
(DCNP)
Saframine
Ninhydrin
Extraction No No Extracted with Chloroform Extraced with Methanol/
Acetone
Heating Time(Min.) 15 min No No No
Absorption maxima at
λmax
570 nm
392 nm with DDQ 640 nm with Methylene
Blue
460 nm with DDQ
425 nm with DCNP 515 nm with saframine
Linearity 50-250 µg/ml 10-80 µg/ml 5-50µg/ml 50-250µg/ml
Asian J. Research Chem. 7(1): January 2014
24
11. Kumaraswami Gandla, JMR Kumar, DVRN B Bikshapati, R
Spandana; A Validated RP-HPLC Method for the Simultaneous
Estimation of Febuxostat and Ketorolac Tromethamine in
Pharmaceutical Pharmulations; Journal of Drug delivery and
Therapeutics; 2013; 2(3); 173-176.
12. Syed Naeem Razzaq, Muhammad Ashfaq, Islam ullah Khan, and
Irfana Marium; Stability Indicating HPLC Method for
Simultaneous Determination of Ofloxacin and Ketorolac
Tromethamine in Pharmaceutical Formulations; Royal Society of
Chemistry; 2012; 4(7); 2121-2126.
13. Padma V. Devrajan, Subhash P. Gore, Sunil V. Chavan; HPTLC
Determination of Ketorolac Tromithamine; Journal of
Pharmaceutical and Biomedical Analysis; 2000; 22(4); 679-683.
14. Rao P.L.K.M., Venugopal V., Teja S.G., Radhika D.V.N.S.,
Kavita K., Lavanya S., Manasa T., Pavani A; Validation and
Analytical Evaluation of Ketorolac Tromethamine by HPTLC
Using Reflectance Scanning Densitometry; International Journal
of Research in Pharmacy and Chemistry; 2011; 1(2); 130-132.
15. Lopez B.E., Castaneda H.G., Gonzalez-De La Parra M., Namur
S.; Development and Validation of High Performance Thin Layer
Cromatographic Method with densitometry for Quantitative
Analysis of Ketorolac Tromethamine in Human Plasma; Journal
of AOAC; 2008; 91(5); 1191-1195.
16. Gandhi S.P., Devani M.G., Borole T.C., Damle M.C.;
Development and Validation of Stability Indicating HPTLC
Method for Determination of Ofloxacin and Ketorolac
Tromethamine in Combination; Journal of Advanced Scientific
Research; 2011; 2(3); 77-82.
17. Patil V.P., Gaikwad A.D., Kulkarni V.S., Kavade R.V., Kale
S.H.; Green Analytical Chemistry; An overview; Inventi Rapid;
Pharma Ana and Qua Assur; 2012; 2;294.
18. Armenta S., Garrigues S., and Della Guardia; Green Analytical
Chemistry; Tr.Ana.Chem.27:M.2008; 497-511.
19. Kamath B.V., Shivram K., Vangani S.; Spectrophotometric
Determination of Ketorolac Tromithamine by Charge Trancefer
and Ion-Pair Complexation; Analytical letters; 1994 ;27(1):103-
112.
20. Pratapreddy A.J. and Chakravarthy I.E., New Spectrophotometric
Determination of Ketorolac Tromithamine Bulk and
Pharmaceutical Dosage Form; International Journal of
Pharmaceutical Science and Research; 2012; 3(12); 4848-4850
... Ketorolac tromethamine works by blocking the production of prostaglandins, compounds that cause pain, fever and inflammation [3]. Ketorolac tromethamine (C 19 H 24 N 2 O 6 ) has a molecular weight 376.4 g/mol and it was estimated by various analytical methods such as spectrophotometry [4][5][6], HPTLC [7,8], voltammetry [9], fluorophotometry [10], HPLC [11][12][13] and in biological fluids such as human plasma [14], human serum [15], human eye samples [16], serum and synovial fluids [17] and post mortem blood samples [18] in the literature. In the present study the authors have proposed five UV spectrophotometric methods for the determination of Ketorolac tromethamine in pharmaceutical dosage forms and the methods were validated as per ICH guidelines [19]. ...
... Literature survey reveals that Ketorolac tromethamine was estimated by various analytical methods such as HPLC [4][5][6], HPTLC [7,8], fluorophotometry [9], voltammetry [10], spectrophotometry [11][12][13] and in biological fluids such as human serum [14], human plasma [15], human eye samples [16], post mortem blood samples ...
... Various spectrophotometric and chromatographic analytical methods are available for determination of both the drugs individual [6][7][8][9][10][11][12][13][14][15][16][17][18][19][20][21][22][23][24][25]. Prava, V. and Seru G. have also reported HPLC method for both the drugs simultaneously [26]. ...
Article
Full-text available
This study involve development of a simple, sensitive, precise, accurate and economic HPLC method and two UV methods for simultaneous estimation of phenylephrine HCl (Phe) and ketorolac tromethamine (Ket). The UV methods are based upon absorption correction and multicomponent analysis approach. Wavelength maxima (λ max) selected for Phe and Ket were 273 nm and 323 nm respectively. Selected linearity ranges for Phe and Ket were 12-42 ppm and 4-14 ppm respectively for UV methods. HPLC method was developed using Waters HILIC (Hydrophilic interaction liquid chromatography) column having dimension of 150 mm x 4.6 mm, 5 μm. Mobile phase used was acetate buffer (pH 3.2; 10mM): MeOH: ACN:: 32:23:45 with flow rate of 1mL/min at detection wavelength of 275nm. Retention time for Phe and Ket were 2.96 and 1.96 min. Linearity ranges for Phe and Ket chosen were 30-60 and 10-20 µg/mL respectively for HPLC method. Thus, selective and robust spectrophotometric and chromatographic methods were developed successfully and also validated for various parameters like accuracy, precision (intra-day and inter-day), linearity and range, limit of detection and limit of quantitation (LOD and LOQ) as per ICH guidelines. %Assay was also determined and found to be in accordance with the label claim. Both the developed methods are quite sensitive for simultaneous identification and analysis of Phe and Ket in bulk. Please cite this article in press as Prof. (Mrs.) Sadhana J. Rajput et al. Simultaneous Estimation of Phenylephrine Hydrochloride and Ketorolac Tromethamine Using UV Spectrophotometric and HPLC Methods.
Article
A cost-effective and simple spectrophotometric technique is established for the quantification of Ofloxacin and Ketorolac tromethamine in bulk drug and eye drops. In the present case the method developed based on simultaneous equation method. The two wavelengths were selected that is 296 nm and 320 nm. The equation was designed and the absorption coefficients for both of the drugs at both the wavelength was calculated. The developed method was applied to formulations too. Analysis of results were statistically validated and by resurgence studies. For routine determination of Ofloxacin and Ketorolac tromethamine, the method was found to be suitable in bulk drug and in formulations. Therefore, the current work aims in developing an easy method for spectrophotometric determination of Ofloxacin and Ketorolac tromethamine in eye drop dose form.
Article
Full-text available
Ketorolac Tromethamine (KT) is prepared for the first time by double emulsion procedure. The current research involves preparation and evaluation of microsponges for ocular applications. This work included preparation of sixteen formulas of KT-microsponges by double emulsion (w/o/w) method using poly lactic-co-glycolic acid (PLGA) as a polymer and poly vinyl alcohol (PVA) as a stabilizer, with different mixer types for different time and power. The prepared microsponges were characterized by Scanning Electron Microscopy (SEM) to investigate the morphology and particle size, the entrapment efficacy and percentage yield were calculated as well as in vitro drug release. Best formula (F14) of KT-microsponges had EE (74%), % yield (83%) with initial drug release (approximately 21% within the first fifteen days) which continued to reach (approximately 86% within 90 days) by using 30% of PLGA concentration with 0.05% of PVA and 200 ml of the external aqueous phase using a probe sonicator for 4 minutes at 200 Watt power. This formulation technique will be the interest of pharmaceutical company.
Article
Full-text available
A spectrophotometric method for the simultaneous estimation of ofloxacin and ketorolac tromethamine in their combined dosage form have been developed and validated for linearity, accuracy, precision, ruggedness and repeatability. The wavelength selected to develop the equation were 300.0 nm and 319.2 nm in acidic methanol. Both drugs shows linearity in the concentration range of 1-11 μg/mL for ofloxacin and 3-13 μg/mL for ketorolac tromethamine. The results have been statistically validated and were found to be simple, rapid, accurate, precise and reproducible.
Article
Full-text available
A simple, precise and accurate method was developed and validated for analysis of Ketorolac Tromethamine in eye drop formulation. An isocratic HPLC analysis was performed on Kromosil C18 column (150 cm × 4.6 mm × 5 μm). The compound was separated with the mixture of methanol and ammonium dihydrogen phosphate buffer in the ratio of 55:45 V/V, pH 3.0 was adjusted with O-phosphoric acid as the mobile phase at flow of 1.5 mL min−1. UV detection was performed at 314 nm using photo diode array detection. The retention time was found to be 6.01 min. The system suitability parameters such as theoretical plate count, tailing and percentage RSD between six standard injections were within the limit. The method was validated according to ICH guidelines. Calibrations were linear over the concentration range of 50–150 μg mL−1 as indicated by correlation coefficient (r) of 0.999. The robustness of the method was evaluated by deliberately altering the chromatographic conditions. The developed method can be applicable for routine quantitative analysis.
Article
Full-text available
A simple, Accurate and precise, selective spectroscopic method was developed and validated for the simultaneous determination of Febuxostat and Ketorolac from bulk and formulations. FBT and KTC were determined at 321nm and 215 nm respectively with the concentration ranges of 2–12mg/ml of FBT and 4-24mg/ml for KTC respectively. The correlation coefficient was found to be 0.9997 & 0.9993 for FBT & KTC drugs at selected wavelengths. The method was validated as per standard analytical procedures, and the results were found to be within limit. Percentage recoveries obtained for both the drugs were 99.65-100.16% and 98.78-99.90%, FBT and KTC respectively. The method was validated according to the ICH guidelines with respect to specificity, linearity, accuracy, precision and robustness. The method developed can be used for the routine analysis of Febuxostat and Ketorolac from their tablet dosage form.
Article
Simple and sensitive spectrophotometric methods are described for the determination of ketorolac tromethamine based on charge-transfer and ion-pair complexation reactions. In the first method the drug was reacted with 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ) or with 2,4-dichloro-6-nitrophenol (DCNP) in methanol. The colour formed due to the formation of charge transfer complex showed maximum absorbance at 392 nm and 425 nm respectively and linearity in the range 10-80 μg.ml. In the second method ketorolac tromethamine was treated with reagents, viz. methylene blue (MB) or saframine (SF), in the presence of phosphate buffer. The ion pair complexes were extracted into chloroform and the absorbance was measured at 640 nm and 515 nm respectively. Beer's law was obeyed in the range of 5-50 μg.ml with both the reagents.* Author for correspondence
Article
We discuss the origins and the fundamentals of Green Analytical Chemistry (GAC), based on the literature published about clean, environmentally-friendly or GAC methods. We pay special attention to the strategies and the tools available to make sample-pretreatment and analytical methods greener. We consider that the main principles are to replace toxic reagents, to miniaturize and to automate methods, making it possible to reduce dramatically the amounts of reagents consumed and wastes generated, so reducing or avoiding side effects of analytical methods. We also consider on-line decontamination or passivation of wastes to be of special interest in making analytical chemistry sustainable.
Article
A simple, specific and accurate stability indicating liquid chromatographic method is established for simultaneous determination of ofloxacin and ketorolac tromethamine in bulk drugs and pharmaceutical formulations. Optimum chromatographic separations among ofloxacin, ketorolac and stress-induced degradation products have been achieved within 15 min by using a BDS Hypersil C8 column (250 × 4.6 mm, 5 μm) as the stationary phase with methanol and phosphate buffer pH 3.0 (55:45 v/v) as the mobile phase at a flow rate of 0.8 mL min−1. Detection is performed at 270 nm using a diode array detector. The method is validated in accordance with ICH guidelines. The response is a linear function of concentration over the range of 12–84 μg mL−1 for ofloxacin and 20–140 μg mL−1 for ketorolac tromethamine. The method results in excellent separation of analytes and their stress induced degradation products with acceptable tailing and resolution. The peak purity index for both the analytes after all types of stress is >0.999 indicating complete separation of both analyte peaks from the stress induced degradation products. The method can therefore be regarded as stability-indicating. The developed method can be applied successfully for simultaneous determination of ofloxacin and ketorolac in pharmaceutical formulations and their stability studies.
Article
Ketorolac tromethamine is a potent nonsteroidal anti-inflammatory drug that is widely used in the treatment of moderate to severe pain. A new method was developed and validated for quantifying ketorolac (the free acid of the tromethamine salt) in human plasma by high-performance thin-layer chromatography. The stationary phase was silica gel 60, and the composition of the mobile phase was n-butanol-chloroform-acetic acid-ammonium hydroxide-water (9 + 3 + 5 + 1 + 2, v/v). The densitometric analysis of ketorolac was performed at 323 nm. The method was validated for precision (repeatability and reproducibility), accuracy, and sensitivity. Repeatability was 10.11% [coefficient of variation (CV)] and reproducibility was 12.18% (CV) as the maximum variation. Accuracy was determined at 3 different concentration levels, and results were within +/-15% of the predetermined range. Data were fitted by a linear mathematical function (linear regression). The calibration graph was linear in the range of 200-2000 ng/mL. Average recovery was 73.67%. The method proved to be accurate, precise, and sensitive for the ketorolac tromethamine quantification.