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Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics



Abdul Rohman, Sudjadi , Devi , Dwiky Ramadhani and Ardi Nugroho
 
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ABSTRACT

FTIR spectroscopy in combination with multivariate calibration of Partial Least Square (PLS) has been developed for quantification of curcumin in the ethanolic extracts of Curcuma longa Linn and Curcuma xanthorriza Roxb. The optimization was done by selecting the best wavenumbers regions capable of providing the high coefficient of determination (R2) and low values of Root Mean Square Error of Calibration (RMSEC). Finally, wavenumbers region of 2000-950 cm–1 was selected for prediction of curcumin in the extracts. The correlation between actual values of curcumin determined by HPLC and FTIR predicted values using FTIR spectroscopy combined with PLS in ethanolic extract of C. longa and C. xanthorriza at 2000-950 cm–1 revealed R2 values of 0.96 and 0.99, respectively. The RMSEC values obtained are 0.299 and 0.089 for C. longa and C. xanthorriza, respectively. The high value of R2 and low value of RMSEC indicated the high accuracy and precision of FTIR spectroscopy for quantification of curcumin in the extracts. These results indicated that FTIR spectroscopy combined with PLS is an alternative technique for determination of curcumin in Curcuma species. The developed method (FTIR spectroscopy) is rapid, no sample preparation and not involving excessive solvents and reagents.

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Abdul Rohman, Sudjadi , Devi , Dwiky Ramadhani and Ardi Nugroho, 2015. Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics. Research Journal of Medicinal Plants, 9: 179-186.

DOI: 10.3923/rjmp.2015.179.186

URL: https://scialert.net/abstract/?doi=rjmp.2015.179.186
 
Received: June 02, 2015; Accepted: July 13, 2015; Published: July 31, 2015



INTRODUCTION

The popularity of herbal medicine has risen, not only in Indonesia having the second largest biodiversity after Brazil but also in the worldwide (Zhang et al., 2012). Curcuma longa known as turmeric is a perennial native plant, where its rhizome is used as a spice, a pigment dye of textiles and in traditional medicine (Jain et al., 2007; Rohman, 2012a). Curcuma longa is one of the plants from Zingiberaceae family and widely cultivated in the regions of tropical and subtropical, especially in India, South East Asia and China. India is the main country exporting the turmeric and its production is approximately 80%. Today, the species cultivation has also widely distributed to some African countries (Parthasarathy et al., 2008). While, Curcuma xanthorriza, known as temu lawak in Indonesian community, is an important and potential medicinal plant belonging to the family Zingiberaceae and shares the same genus as Curcuma longa Linn (Ab Halim et al., 2012).

The main compounds contributing to the activities of C. longa Linn and C. xanthorriza Roxb. are curcuminoids, including curcumin, demethoxycurcumin and bis-demethoxycurcumin (Fig. 1). Among these, curcumin is present as major component in C. longa and C. xanthorriza.

Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics
Fig. 1(a-c): Chemical structure of, (a) Curcumin, (b) Demethoxycurcumin and (c) Bis-demethoxycurcumin

There are several reports regarding the pharmacological activities of curcumin, such as its anti-inflammatory, antimicrobial, antioxidant, antiparasitic, antimutagenic and anticancer properties (Kalpravidh et al., 2010; Skrzypczak-Jankun et al., 2000; Singh et al., 2002; Aggarwal et al., 2003). It is also efficient in the treatment of liver diseases and dermatological disorders (Semwal et al., 1997).

Some analytical methods, including High Performance Liquid Chromatography (HPLC) and its coupling to mass spectrometry (LC-MS) (Bos et al., 2007; Jiang et al., 2006), Thin Layer Chromatography (TLC) (Phattanawasin et al., 2009; Paramasivam et al., 2009) and capillary electrophoresis (Lechtenberg et al., 2004) have been used to analyze the curcuminoids in various turmeric samples. However, such chromatographic methods are time-consuming, require experienced personnel to perform the analysis and are destructive. Therefore, some nondestructive and reliable techniques based on vibrational spectroscopy have been developed for quantitative analysis such as near infrared spectroscopy and UV spectrophotometer (Tanaka et al., 2008; Pothitirat and Gritsanapan, 2006).

Fourier Transform Infrared (FTIR) spectroscopy, especially in combination with chemometrics technique, has been widely used in analysis of herbal medicine (Rohman et al., 2014). The method allows fast, easein sample preparation and non destructive technique. In addition, FTIR spectroscopy can be exploited for determination of components on interesting herbal medicine simultaneously. Using literature review, there is no report regarding the use of FTIR spectroscopy for quantification of curcumin in C. longa and C. xanthorriza, therefore, this present study aimed to evaluate the application of FTIR spectroscopy in quantitative analysis of curcumin in ethanolic extracts of C. longa and C. xanthorriza.

MATERIALS AND METHODS

The standard of curcumin was obtained from synthesis and kindly given by Prof. Dr. Sudibyo Martono from Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Gadjah Mada University, Indonesia. The curcuminoids, i.e., the mixture of curcumin, demethoxycurcumin and bis-demethoxycurcumin were obtained from Sigma (Alldrich, St. Louis, USA) with the purity of >99%. Acetonitrile and methanol used were of HPLC grade. KBr used for FTIR spectroscopy sampling preparation was purchased from Sigma (Alldrich, St. Louis, USA).

Plant materials and extraction: The rhizomes of C. longa and C. xanthorriza were obtained from several traditional markets in central java and Daerah Istimewa Yogyakarta, Indonesia in September-October 2013. All rhizome samples were cleaned, cut and air-dried and finally powdered. The powder of C. longa and C. xanthorriza was subjected to extraction using maceration technique using ethanol as solvent to obtain ethanolic extract of C. longa and C. xanthorriza, respectively. These extracts were used for FTIR spectroscopic measurement.

FTIR spectral analysis: A 50 mg ethanolic extract of C. longa and C. xanthorriza was added with 950 mg KBr IR grade and grinded in mortar until homogen. The mixture was placed on Horizontal Attenuated Total Reflectance (HATR) accessory at controlled ambient temperature (20°C). The FTIR spectra of all samples were scanned using a FTIR spectrophotometer ABB MB3000 (Clairet Scientific, Northampton, UK), equipped with deuterated triglycine sulphate (DTGS) detector and beam splitter of germanium. Using Horizon MB FTIR software version 3.0.13.1 (ABB, Canada) included in the instrument, FTIR spectra were scanned in the mid infrared region of 4000-650 cm–1 with resolution of 4 cm–1 and number of scanning of 32. The samples were placed in good contact with HATR accessory. All spectra were rationed against a background of air spectrum. After every scan, a new reference air background spectrum was taken. These spectra were recorded as absorbance values at each data point in triplicate.

Quantitation of curcuminoids by High Performance Liquid Chromatography (HPLC): The contents of curcumin in ethanolic extracts of C. longa and C. xanthorriza determined by HPLC were used as actual value or reference value for those obtained with FTIR spectroscopy in combination with multivariate calibration of Partial Least Square (PLS). The extract was dissolved in mobile phase and an aliquot of 2.0 mL is taken, filtered through an HPLC filter and placed in an autosampler vial of 2 mL. The HPLC analysis was performed using Waters HPLC system (Waters Corp., USA), consisting pump (Waters), ACQUITY solvent manager, waters alliance column heater, Vial Amber Glass 12×32 mm 2 mL with Cap and PTFE/silicone Septum, waters 2767 sample manager and operating software of Empower Basic 2 (Waters, USA). Separation of curcuminoids is performed using C18 WatersXterra MS C18 (5 μm; 4, 6×250 mm). The mobile phase used consisted of aquabidestilata and acetonitrile (65:35 v/v) containing 1% acetic acid. The analyte detection was done using UV-vis detector set at λ 425 nm. The injection volume was 20 μL and running time was 40 min.

Statistical analysis: The multivariate calibration of PLS for the correlation between actual value of curcumin as determined with HPLC and FTIR predicted value was performed using Horizon software included in FTIR spectrophotometer. The leave-one-out cross-validation procedure was used to verify the calibration model. The values of Root Mean Square Error of Calibration (RMSEC) and coefficient of determination (R2) were used as the validity criteria for calibration model. The predictive ability of PLS calibration model was further used to calculate the validation or prediction samples.

RESULTS AND DISCUSSION

Quantification of curcumin by HPLC: Curcumin in ethanolic extracts of C. longa and C. xanthorriza was initially determined using high performance liquid chromatography with UV-vis detector. Figure 2 exhibited the example of HPLC chromatogram during separation of curcumin from other two curcuminoids using condition as above, either in curcuminoids obtained from Mercck or in ethanolic extract of C. xanthorriza. The retention times are 28.191, 31.757 and 35.688 min for bisdes methoxycurcumin, desmethoxycurcumin and curcumin, respectively. Curcuma xanthorriza contained curcumin and bidesmethoxycurcumin, as represented by peaks having retention times close to curcumin and bisdesmethoxycurcumin.

Table 1 compiled the levels of curcumin in some ethanolic extract of C. longa. While, the level of curcumin in ethanolic extract of C. xanthorriza was compiled in Table 2. The levels of curcumin in the evaluated samples are diverse due to the different region, age of plants, etc. Comparing Table 2 and 3, it is known that curcumin concentration in C. longa is higher than that in C. xanthorriza, as reported by several investigators (Lechtenberg et al., 2004; Bos et al., 2007; Li et al., 2011).

Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics
Fig. 2(a-b):
The HPLC chromatogram of curcuminoids from (a) E. Merck and (b) Ethanolic extract of Curcuma xanthorriza. The retention times for bis desmethoxycurcumin, desmethoxycurcumin and curcumin are 28.191, 31.757 and 35.688 min, respectively.

Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics
Fig. 3:The FTIR spectra of ethanolic extract of Curcuma longa L. scanned at mid infrared region (4000-6500 cm–1). X-axis: Wavenumbers and Y-axis: Response (Absorbance)

Table 1:Levels of curcumin in ethanolic extract of Curcuma longa in some regions
Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics

Table 2:Levels of curcumin in ethanolic extract of Curcuma xanthorriza in some regions
Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics

The level of curcumin in ethanolic extract of C. longa varied from 4.78±2.25-7.36±0.09% wt/wt, while ethanolic extract of C. xanthorriza has curcumin level of 1.66±0.01-2.97±0.05% wt/wt. These values were further used as actual values during correlation with values of curcumin levels obtained using FTIR spectroscopy and multivariate calibration of PLS.

Analysis of curcumin using FTIR spectroscopy and multivariate calibration: Figure 3 revealed FTIR spectra of ethanolic extract of C. longa (turmeric) from different regions. Each peak and shoulders come from the absorption of functional groups in C. longa extract. Investigation of FTIR spectra of all ethanolic extract of C. longa clearly shows the similar peaks due to the similar chemical components contained. However, using detail scrutiny, there is a bit difference in terms of peak intensity (absorbance) caused by different concentrations of components present in ethanolic extract of C. longa.

Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics
Fig. 4:
Correlation between actual values of curcumin in ethanolic extract of Curcuma longa determined by HPLC method (x-axis) and predicted values using FTIR spectroscopy combined with PLS (y-axis) at wavenumbers of 2000-950 cm–1

Table 3:Functional groups responsible for infrared absorption of ethanolic extract of Curcuma longa
Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics

The main components of ethanolic extract of C. longa is curcuminoids which refer to phenolic compounds responsible for yellow color in turmeric. As consequence, there is no surprising if peaks coming from functional groups of curcuminoids dominate FTIR spectra of ethanolic extract of turmeric. The functional groups responsible for infrared absorption of ethanolic extract of C. longa was compiled in Table 3.

The first step for analysis of curcumin in C. longa and C. xanthorriza using FTIR spectroscopy combined with multivariate calibration of Partial Least Square (PLS) is the selection of wavenumbers region. The use of FTIR spectra at selected regions can increase the accuracy of analytical results (Vazquez et al., 2000). The selection of wave numbers region is based on its capability to provide the high coefficient of determination (R2) and low values of errors, either in calibration model known as Root Mean Square Error of Calibration (RMSEC) or in prediction model called with Root Mean Square Error of Prediction (RMSEP) (Rohman, 2012b). After optimization step, finally, wave numbers region of 2000-950 cm–1 was selected for prediction of curcumin.

Figure 4 and 5 reveled the correlation between actual values of curcumin determined by HPLC and FTIR predicted values using FTIR spectroscopy combined with PLS at wavenumbers of 2000-950 cm–1 in ethanolic extract of C. longa and C. xanthorriza. The R2 values obtained are 0.96 and 0.99 in C. longa and C. xanthorriza, respectively. The RMSEC values obtained are 0.299 and 0.089. The high value of R2 and low value of RMSEC indicated the high accuracy and precision of analytical method.

Image for - Analysis of Curcumin in Curcuma longa and Curcuma xanthorriza Using FTIR Spectroscopy and Chemometrics
Fig. 5:
Correlation between actual values of curcumin in ethanolic extract of C. xanthorriza determined by HPLC method (x-axis) and predicted values using FTIR spectroscopy combined with PLS (y-axis) at wavenumbers of 2000-950 cm–1

CONCLUSION

These results indicated that FTIR spectroscopy combined with PLS is an alternative technique for determination of curcumin in Curcuma species.

ACKNOWLEDGMENT

We acknowledge to Directorate of higher Education, The Ministry of Research, Technology and Higher Education for financial supporting during this research via Comprehensive Research Grant awarded to Dr. Abdul Rohman.

REFERENCES

1:  Ab Halim, M.R., M.S.M.Z. Tan, S. Ismail and R. Mahmud, 2012. Standardization and phytochemical studies of Curcuma xanthorrhiza Roxb. Int. J. Pharm. Pharmaceut. Sci., 4: 606-610.
Direct Link  |  

2:  Aggarwal, B.B., A. Kumar and A.C. Bharti, 2003. Anticancer potential of curcumin: Preclinical and clinical studies. Anticancer Res., 23: 363-398.
PubMed  |  Direct Link  |  

3:  Bos, R., T. Windono, H.J. Woerdenbag, Y.L. Boersma, A. Koulman and O. Kayser, 2007. HPLC-photodiode array detection analysis of curcuminoids in Curcuma species indigenous to Indonesia. Phytochem. Anal., 18: 118-122.
CrossRef  |  Direct Link  |  

4:  Jain, S., S. Shrivastava, S. Nayak and S. Sumbhate, 2007. Recent trends in Curcuma longa Linn. Pharmacogn. Rev., 1: 119-128.
Direct Link  |  

5:  Jiang, H., B.N. Timmermann and D.R. Gang, 2006. Use of liquid chromatography-electrospray ionization tandem mass spectrometry to identify diarylheptanoids in turmeric (Curcuma longa L.) rhizome. J. Chromatogr. A, 1111: 21-31.
CrossRef  |  Direct Link  |  

6:  Kalpravidh, R.W., N. Siritanaratkul, P. Insain, R. Charoensakdi and N. Panichkul et al., 2010. Improvement in oxidative stress and antioxidant parameters in β-thalassemia/Hb E patients treated with curcuminoids. Clin. Biochem., 43: 424-429.
CrossRef  |  Direct Link  |  

7:  Li, R., C. Xiang, M. Ye, H.F. Li, X. Zhang and D.A. Guo, 2011. Qualitative and quantitative analysis of curcuminoids in herbal medicines derived from Curcuma species. Food Chem., 126: 1890-1895.
CrossRef  |  Direct Link  |  

8:  Lechtenberg, M., B. Quandt and A. Nahrstedt, 2004. Quantitative determination of curcuminoids in Curcuma rhizomes and rapid differentiation of Curcuma domestica Val. and Curcuma xanthorrhiza Roxb. by capillary electrophoresis. Phytochem. Anal., 15: 152-158.
CrossRef  |  Direct Link  |  

9:  Paramasivam, M., R. Poi, H. Banerjee and A. Bandyopadhyay, 2009. High-performance thin layer chromatographic method for quantitative determination of curcuminoids in Curcuma longa germplasm. Food Chem., 113: 640-644.
CrossRef  |  Direct Link  |  

10:  Phattanawasin, P., U. Sotanaphun and L. Sriphong, 2009. Validated TLC-image analysis method for simultaneous quantification of curcuminoids in Curcuma longa. Chromatographia, 69: 397-400.
CrossRef  |  Direct Link  |  

11:  Parthasarathy, V.A., B. Chempakam and T.J. Zachariah, 2008. Chemistry of Spices. CABI Press, Kingslynn, UK., ISBN-13: 978-1845934057, pp: 97-98

12:  Pothitirat, W. and W. Gritsanapan, 2006. Variation of bioactive components in Curcuma longa in Thailand. Curr. Sci., 91: 1397-1400.
Direct Link  |  

13:  Rohman, A., 2012. Analysis of curcuminoids in food and pharmaceutical products. Int. Food Res. J., 19: 59-66.
Direct Link  |  

14:  Rohman, A., 2012. Application of Fourier transform infrared spectroscopy for quality control of pharmaceutical products: A review. Indonesian J. Pharm., 23: 1-8.
Direct Link  |  

15:  Rohman, A., A. Nugroho, E. Lukitaningsih and Sudjadi, 2014. Application of vibrational spectroscopy in combination with chemometrics techniques for authentication of herbal medicine. Applied Spectrosc. Rev., 49: 603-613.
CrossRef  |  Direct Link  |  

16:  Semwal, A.D., G.K. Sharma and S.S. Arya, 1997. Antioxygenic activity of turmeric (Curcuma longa) in sunflower oil and ghee. J. Food Sci. Technol., 34: 67-69.
Direct Link  |  

17:  Singh, R., R. Chandra, M. Bose and P.M. Luthra, 2002. Antibacterial activity of Curcuma longa rhizome extract on pathogenic bacteria. Curr. Sci., 83: 737-740.
Direct Link  |  

18:  Skrzypczak-Jankun, E., N.P. McCabe, S.H. Selman and J. Jankun, 2000. Curcumin inhibits lipoxygenase by binding to its central cavity: Theoretical and X-ray evidence. Int. J. Mol. Med., 6: 521-527.
CrossRef  |  PubMed  |  Direct Link  |  

19:  Tanaka, K., Y. Kuba, T. Sasaki, F. Hiwatashi and K. Komatsu, 2008. Quantitation of curcuminoids in curcuma rhizome by near-infrared spectroscopic analysis. J. Agric. Food Chem., 56: 8787-8792.
CrossRef  |  Direct Link  |  

20:  Vazquez, P.P., M.M. Galera, A.G. Frenich and J.L.M. Vidal, 2000. Comparison of calibration methods with and without feature selection for the analysis of HPLC data. Anal. Sci., 16: 49-55.
CrossRef  |  Direct Link  |  

21:  Zhang, J., B. Wider, H. Shang, X. Li and E. Ernst, 2012. Quality of herbal medicines: Challenges and solutions. Complement. Ther. Med., 20: 100-106.
CrossRef  |  Direct Link  |  

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