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Asian Journal of Biomaterial Research 2017; 3(4):14-18 14
Research Article
UV Spectrophotometric Method Development and Validation for Quantitative Estimation of
Curcumin
Sagar Kishor Savale*
*Department of Pharmaceutics, R. C. Patel Institute of Pharmaceutical Education and Research,
Shirpur 425-405, MS, India
Received: 02 August 2017 Revised: 6 August 2017 Accepted: 10 August 2017
Abstract
Aim: Development and Validation of a method for the UV determination of Curcumin.
Objective: U.V Spectrophotometric method have been widely employed in determination of
Curcumin in a mixture or fixed dose combination. For the ternary mixture containing Curcumin, no
spectrophotometric method for evaluation has been reported so far. Thus our aim is to develop
method for Curcumin estimation in ternary mixture using U.V spectrophotometry.
Method: The method was validated as per ICH guidelines. The recovery studies confirmed the
accuracy and precision of the method.
Conclusion: It was successfully applied for the analysis of the drug in bulk as well as biological
samples and could be effectively used for the routine analysis.
Key words: Curcumin, UV spectra, method development, Validation, ICH.
Introduction CRM is a bright yellow-orange powder
Curcumin ((1E, 6E)-1, 7-Bis (4-hydroxy-3- material and CRM having maximum solubility
methoxyphenyl)-1, 6 heptadiene-3, 5-dione), a in methanol and acetonitrile. Melting point of
polyphenol known as diferuloylmethane. CRM was 185ºc and λmax of CRM is 423 nm
Molecular formula of curcumin (CRM) C21 respectively.
H20 O6 and molecular weight of CRM is CRM was identified by Melting point, infrared
368.39 g/mol. CRM is highly lipophilic drug vibrational spectrophotometry (IR),
having a log P value is 1.82 and dissociation Differential scanning Calorimetry
constant of CRM was 8.3±0.04 respectively (DSC).CRM are effective chemotherapeutic
(Han et al., 2003). agents against a wide variety of cancer types.
CRM enhances cytotoxicity via
*
Corresponding author, downregulation of nuclear factor (NF)-ĸB and
Mr. Sagar Kishor Savale, the Akt pathways, thereby reversing MDR.
Department of Pharmaceutics, This study was aimed at developing a simple,
R. C. Patel Institute of Pharmaceutical rapid and sensitive method for estimation of
Education & Research, Shirpur, 425405, analyte (CRM) in bulk samples by using UV
dist. Dhule, Maharashtra, India. spectrophotometric method (Andriamanana et
Mobile No: +91 9960885333, al., 2013; Couchman et al., 2012).
Email ID: avengersagar16@[Link]
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Asian Journal of Biomaterial Research 2017; 3(4):14-18 15
Material and Method (2-12 µg/mL) and examined between 800-200
Material nm. The maximum absorbance was
Curcumin supplied as a gift sample by determined using UV-Vis Specrophotometer
Sunpure Extracts Pvt. Ltd (Delhi, India) used (UV-1700, Shimadzu, Japan) to confirm the
as working standard. λmax of the drugs.
Instrumentation Validation of analytical method
A double beam UV-VIS spectrophotometer The analytical performance characteristics
(UV-1700, Shimadzu, Japan) connected to a which may be tested during methods
computer loaded with spectra manager validation: % Recovery, Precision,
software UV Probe was used. The spectra Ruggedness and sensitivity (Dangre et al.,
were obtained with the instrumental 2015; Savale et al., 2017).
parameters as follows: Wavelength range: Results and Discussion
800-200 nm. All weights were taken on an Method Development
electronic balance (Model Shimadzu AUX The solution of CRM in methanol was found
120). to exhibit maximum absorption at 423 nm
Preparation of standard stock solution after scanning on the UV-Vis
According to European pharmacopoeia, 10 mg spectrophotometer which was reported as
of CRM was dissolve in 100 ml of methanol λmax in the literature and the procured drug
(100 µg/mL). Out of this stock 0.2-1.2 ml was sample of CRM complies with the reference
pipetted and diluted up to 10 ml by methanol spectra (Figure 1).
Figure 1. UV spectra of Curcumin (CRM)
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Asian Journal of Biomaterial Research 2017; 3(4):14-18 16
Linearity study made. Absorbance of the above solution were
Accurately weighted CRM (10 mg) was taken at 423 nm by using UV-Vis
dissolved in 100 ml of methanol to obtain spectrophotometer (UV-1700, Shimadzu,
working standard of 100 μg/ml. Aliquots were Japan) against the blank solution prepared in
pipetted from the stock solution of drug and the same manner without adding the drug. A
were transferred to 10 ml volumetric flask, the graph of absorbance vs concentration was
final volume was adjusted with methanol so plotted (Figure 2) and R2 was found to be
that concentration of 2-12 μg/ml could be 0.9999.
Figure 2. Calibration curve of CRM
Validation of analytical method
Recovery concentration levels i.e 80%, 100% of assay
Recovery study is performed by standard concentration and % recovery for all these
addition method by adding the known amount drug were calculated. Result was reported in
of CRM (Working standard) at two different Table 1.
Table 1. Recovery study
Drug Initial amount Added Amount % % RSD
(µg/ml) (µg/ml) Recovery (n = 3)
CRM 2 2 101.05 0.04
2 2 105.94 0.09
Precision day variability was assessed using above
Intra-day precision was determined by mentioned two concentrations analysed on
analysing, the two different concentrations 2 three different days, over a period of one week
mg/ml, 3 mg/ml containing CRM, for three (n = 3) Table 2.
times in the same day (n = 3) Table 2. Inter-
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Asian Journal of Biomaterial Research 2017; 3(4):14-18 17
Table 2. Presion study
Intra - Day Inter - Day
Drug Con. (µg/ml) Mean ± SD % RSD Mean ± SD % RSD
CRM 2 2.0 ± 0.0015 0.05 2.0 ± 0.0009 0.07
3 3.0 ± 0.0010 0.03 3.0 ± 0.0033 0.09
Ruggedness similar operational and environmental
From stock solution, sample solution conditions (Table 3) (n = 3).
containing CRM (2 µg/ml) was prepared and
analyzed by two different analysts using
Table 3. Ruggedness study
% Amount Found % RSD
Drug Analyst I Analyst II Analyst I Analyst II
CRM 100.00 100.06 0.01 0.03
Sensitivity (LOD) and Limit of Quantitation (LOQ)
Sensitivity of the proposed method were (Table 4).
estimated in terms of Limit of Detection
Table 4. Sensitivity study
Drug LOD LOQ
CRM 0.38 ± 0.003 0.99 ± 0.016
Conclusion for the measurement of a range of
The proposed UV spectrophotometric method tyrosine kinase inhibitors in human
was found very simple, rapid and economical. plasma or serum using turbulent flow
The method is validated in compliance with liquid chromatography–tandem mass
ICH guidelines is suitable for estimation of spectrometry. Anal Bioanal Chem., 403:
CRM with excellent recovery, precision and 1685-1695.
linearity. Dangre P, Sawale V, Meshram S, Gunde M.
2015. Development and validation of RP-
Reference HPLC method for the Simultaneous
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LC/MS/MS assay for curcumin and Spectrophotometric Method Development
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