Malaysian Journal of Analytical Sciences Vol 23 No 4 (2019): 715 – 724

DOI: 10.17576/mjas-2019-2304-18

 

 

 

QUANTITATIVE ANALYSIS OF PHENOLICS CONTENT IN TWO ROSELLE VARIETIES (Hibiscus sabdariffa) BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY

 

(Analisis Kuantitatif bagi Sebatian Fenolik dalam Dua Varieti Roselle (Hibiscus sabdariffa) menggunakan Kromatografi Cecair Berprestasi Tinggi)

 

Siti Aishah Mohd Ali^1,2,  Satirah Zainalabidin³,  Jalifah Latip¹*

 

¹Chemistry Programme, Faculty of Science and Technology,

Universiti Kebangsaan Malaysia, 43600 UKM Bangi, Selangor, Malaysia

²Faculty of Science and Natural Resources,

Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia

³School of Applied Health, Faculty of Health Sciences,

Universiti Kebangsaan Malaysia, 50300 Kuala Lumpur, Malaysia

 

*Corresponding author:  jalifah@ukm.edu.my

 

Received: 31 March 2018; Accepted: 17 April 2019

 

 

Abstract

Phenolics content of two roselle varieties, H. sabdariffa var. UKMR-2 and H. sabdariffa var. UMKL-1 was determined based on high performance liquid chromatography method using a photodiode array detector (HPLC-PDA) with gradient elution. Roselle calyces were extracted with water via sonication (50 ^°C, 30 minutes). The chromatographic separation was carried out on Purospher STAR RP-18e LichroCART column (250 mm × 4.6 mm × 5 µm) and detection was carried out at three different wavelengths (265, 320 and 520 nm). The developed HPLC method provided high percentage of recovery (95 - 102%), good linearity at R² > 0.99, low limit of detection (LOD) and low limit of quantitation (LOQ). Two predominant anthocyanins were detected by HPLC-PDA in both H. sabdariffa varieties and were identified as delphinidin-3-O-sambubioside and cyanidin-3-O-sambubioside. In addition, chlorogenic acid, caffeic acid and ascorbic acid were also detected. High content of phenolics and ascorbic acid were detected in UKMR-2 as compared to UMKL-1, suggested this variety have high potential for commercial cultivation.

 

Keywords:  anthocyanins, Hibiscus sabdariffa, HPLC-PDA

 

Abstrak

Kandungan fenolik bagi dua varieti rosel, H. sabdariffa var. UKMR-2 dan H. sabdariffa var. UMKL-1 ditentukan berdasarkan kaedah kromatografi cecair berprestasi tinggi menggunakan pengesan foto-diod (KCPT-PFD) dengan elusi kecerunan kekutuban. Kaliks rosel diekstrak dengan air secara sonikasi (50 °C, 30 minit). Pemisahan kromatografi dilakukan menggunakan turus Purospher STAR RP-18e LichroCART (250 mm × 4.6 mm × 5 µm) dan pengesanan dilakukan pada tiga panjang gelombang yang berbeza (265, 320 dan 520 nm). Kaedah KCPT yang dibangunkan mempunyai peratusan dapatan semula yang tinggi (95 - 102%), lineariti yang baik pada R² > 0.99 serta had pengesanan (LOD) dan kuantifikasi (LOQ) yang rendah. Dua sebatian antosianin dikesan oleh KCPT-PFD dalam kedua-dua varieti H. sabdariffa dan dikenal pasti sebagai delfinidin-3-O-sambubiosida dan sianidin-3-O-sambubiosida. Selain itu, asid klorogenik, asid kafeik dan asid askorbik juga berjaya dikesan. UKMR-2 dikesan mengandungi sebatian fenolik dan asid askorbik yang tinggi berbanding UMKL-1, mencadangkan varieti ini mempunyai potensi yang tinggi sebagai tanaman komersil.

 

Kata kunci:  antosianin, Hibiscus sabdariffa, KCPT-PFD

 

References

1.       Padmaja, H., Sruthi, S. and Vangalaapati, M. (2014). Review on Hibiscus sabdariffa – A valuable herb. International Journal of Pharmacy & Life Sciences, 5(8): 3747-3752.

2.       Fasoyiro, S.B., Babalola, S.O. and Owosibo T. (2005). Chemical composition and sensory quality of fruit-flavoured roselle (Hibiscus sabdariffa) drinks. World Journal of Agricultural Sciences, 1(2): 161-164.

3.       Puro, K., Sunjukta, R., Samir, S., Ghatak, S., Shakuntala, I. and Sen, A. (2014). Medicinal uses of roselle plant (Hibiscus sabdariffa L.): A mini review. Indian Journal of Hill Farming, 27(1): 81-90.

4.       Azza, A., Ferial, M. and Esmat, A. (2011). Physicochemical properties of natural pigments (anthocyanin) extracted from Roselle calyces (Hibiscus sabdariffa). Journal of American Science, 7(7): 445-456.

5.       Obouayeba, A. P., Djyh, N. B., Diabate, S., Djaman, A. J., N’guessan, J. D., Kone, M. and Kouakou, T. H. (2014). Phytochemical and antioxidant activity of roselle (Hibiscus sabdariffa L.) petal extracts. Research Journal of Pharmaceutical, Biological and Chemical Sciences, 5(2): 1453-1465.

6.       Osman, M., Golam, F., Saberi, S., Majid, N. A., Nagoor, N. H. and Zulqarnain. M. (2011). Morpho-agronomic analysis of three roselle (Hibiscus sabdariffa L.) mutants in tropical Malaysia. Australian Journal of Crop Science, 5(10):1150-1156.

7.       Sukwattanasinit, T., Burana-osot, J. and Sotanaphun, U. (2007). Spectrophotometric method for quantitative determination of total anthocyanins and quality characteristics of roselle (Hibiscus sabdariffa). Planta Medica, 73: 1517-1522.

8.       Kouakou, T. H., Konkon, N. G., Ayolie, K., Obouayeba, A. P., Abeda, Z. H. and Kone, M. (2015). Anthocyanin production in calyx and callus of roselle (Hibiscus sabdariffa L.) and its impact on antioxidant activity. Journal of Pharmacognosy and Phytochemistry, 4(3): 9-15.

9.       Bernal, F. A., Orduz-Diaz, L. L. and Coy-Barrera, E. (2016). Application of FARAFAC and OPLS-DA analysis on HPLC fingerprints for the characterization of Hibiscus sabdariffa calyces. Quimica Nova, 39(2): 160-166.

10.    Rodreguez-Medina, I.C., Debon, R.B., Molina, V.M., Villaverde, C.A., Joven, J., Menendez, J.A., Carretero, A.S. and Gutierrez, A.F. (2009). Direct characterization of aqueous extract of Hibiscus sabdariffa using HPLC with diode array detection coupled to ESI and ion trap MS. Journal of Separation Science, 32: 3441-3448.

11.    Diniz, A. C. B., Astarita, L.V. and Santaré, E. R. (2007). Alterac¸ ão dos metabólitos secundáriosem plantas de Hypericum perforatum L. (Hypericaceae) submetidas à secagem eao congelamento. Acta Botanica Brasilica, 21, 443–450.

12.    Gobbo-Neto, G. L. and Lopes, N. P. (2007). Plantas medicinais: fatores de influência noconteúdo de metabólitos secundários. Quimica Nova, 30, 374-381.

13.    Naczk, M. and Shahidi, F. (2004). Extraction and analysis of phenolics in food. Journal of Chromatography, 1054: 95-111.

14.    Chumsri, P., Airichote, A. and Itharat, A. (2008). Studies on the optimum conditions for the extraction and concentration of roselle (Hibiscus sabdariffa Linn.) extract. Songklanakarin Journal of Science and Technology, 30(1): 133-139.

15.    Validation of analytical procedures: Text and methodology Q2 (R1). (2005). ICH Harmonised Tripartite Guideline, International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use.  Access from http://www.ich.org/fileadmin/Public_Web_Site/ICH_Products/Guidelines/Quality/Q2_R1/Step4/Q2_R1_Guideline.pdf. [Access online 10 October 2017].

16.    Anvisa (2003). Regulation-RE no. 899: Guide for validation of analytical and bioanalytical methods. National Health Surveillance Agency, Brazil. http://www.in.gov.br/imprensa/visualiza/index. [Access online 10 October 2017].

17.    Chin, K. L., Zhen, J., Qi, Y., Chin, S. L., Breithaupt, M., Wu, Q. L., Simon, J., Henson, J. and Ferchaud, V. (2016). A comparative evaluation: Phytochemical composition and antioxidant capacity of three roselle (Hibiscus sabdariffa L.) accessions. Acta Horticulturae, 1125: 99-108.

18.    Ifie, I., Ifie, B. E., Ibitoyed, D. O., Marshalla, L. J. and Williamson, G. (2018). Seasonal variation in Hibiscus sabdariffa (roselle) calyx phytochemical profile, soluble solids and α-glucosidase inhibition. Food Chemistry, 261: 164-168.

19.    Amor, B. B. and Allaf, K. (2009). Impact of texturing using instant pressure drop treatment prior to solvent extraction of anthocyanins from Malaysian roselle (Hibiscus sabdariffa). Food Chemistry 115(3): 820-825.

20.    Ichiyanagi, T., Shida, Y., Rahman, M. M., Hatano, Y. and Konishi, T. (2006). Bioavailability and tissue distribution of anthocyanins in bilberry (Vaccinium myrtillus L.) extract in rats. Journal of Agriculture Food Chemistry, 54: 6578–6587.

21.    Deshmukh, S. R., Wadegaonkar, V. P., Bhagat, R. P. and Wadegaonkar, P. A. (2011). Tissue specific expression of anthraquinones, flavonoids and phenolics in leaf, fruit and root suspension cultures of Indian mulberry (Morinda citrifola L.). Plant Omics Journal, 4: 6-13.

22.    Biswas, N., Balac, P., Narlakanti, S. K., Enamul Haque, M. D. and Mehedi Hassan, M. D. (2013). Identification of phenolic compounds in processed cranberries by HPLC method. Journal of Nutrition and Food Science, 3: 181.

23.    Alarcon-Aguilar, F. J., Zamilpa, A., Perez-Garcia, M. D., Almanza-Perez, J. C., Romero-Nunez, E. and Campos-Sepulveda, E. A. (2007). Effect of Hibiscus sabdariffa on obesity in MSG mice. Journal of Ethnopharmacology, 114(1): 66-71.

24.    Bamishaiye, E. I., Olayemi, F. F. and Bamishaiye, O. M. (2011). Effects of boiling time on mineral and vitamin C content of three varieties of Hibiscus sabdriffa drink in Nigeria. World Journal of Agricultural Sciences, 7(1): 62-67.

25.    Prabhakar, B. and Pandita, N. (2010). Quantitative HPLC analysis of ascorbic acid and gallic acid in Phyllanthus emblica. Journal of Analytical and Bioanalytical Techniques, 1: 111.

26.    Assis, S. A., Oliveira, O. M. M. F. and Lima, D. C. (2001). Activity of pectin methylesterase, pectin content and vitamin C in acerola fruit at various stages of fruit development. Food Chemistry, 74: 133-136.

27.    Lee, J. K. and Kader, A. A. (2000). Pre-harvest and post-haverst factor influencing vitamin C content of horticultural crops. Post-harvest Biology and Technology, 20: 207-220.

28.    Bouayed, J., Rammal, H., Dicko, A., Younos, C. and Soulimani, R. (2007). Chlorogenic acid, a polyphenol from Prunus domestica (Mirabelle), with coupled anxiolytic and antioxidant effects. Journal of the Neurological Sciences, 262: 77-84.

29.    Lan, W. U. (2007). Effect of chlorogenic acid on antioxidant activity of Flos lonicerae extracts. Journal of Zhejiang University Science, 8(9): 673-679.

30.    Seal, T. (2016). Quantitative HPLC analysis of phenolic acids, flavonoids and ascorbic acid in four different solvent extracts of two wild edible leaves, Sonchus arvensis and Oenanthe linearis of North-Eastern region in India. Journal of Applied Pharmaceutical Science, 6(2): 157-166.

31.    Juliani, H. R., Welch, C. R., Wu, Q., Diouf, B., Malainy, D. and Simon, J. E. (2009). Chemistry and quality of Hibiscus (Hibiscus sabdariffa) for developing the natural-product industry in Senegal. Journal of Food Science, 74: S113.

32.    Khafaga, E. R. and Koch, H. (1980). Stage of maturity and quality of roselle (Hibiscus sabdariffa L. var. sabdariffa) - Organic acids. Angewandte Botanik, 54: 287-293.

33.    Hassanein, R. A., Khattab, H. K. I., EL-Bassiouny, H. M. S. and Sadak, M. S. (2005). Increasing the active constituents of sepals of roselle (Hibiscus sabdariffa L.) plant by applying gibberellic acid and benzyladenine. Journal of Applied Sciences Research, 1(2): 137-146.

34.    Macheix, J. J., Fleuriet, A. and Billot, J. (1990). Fruit Phenolic. Boca Raton, CRC.

35.    Buta, J. G. and Spaulding, D.W. (1997). Endogenous levels of phenolics in tomato fruit during growth and maturation. Journal of Plant Growth Regulation, 16: 43-46.

36.    Raffo, A., Cherubino, L., Vincenzo, F., Ambrozino, P., Salucci, M., Gennaro, L., Bugianesi, R., Giuffrida, F. and Quaglia, G. (2002). Nutritional value of cherry tomatoes (Lycopersicon esculentum cv. Naomi F1) harvested at different ripening stages. Journal of Agricultural and Food Chemistry, 50: 6550-6556.