Malaysian Journal of Analytical Sciences Vol 22 No 2 (2018): 219 - 226

DOI: 10.17576/mjas-2018-2202-06

 

 

 

VALIDATED RP-HPLC METHOD FOR QUANTIFICATION OF GASTRODIN IN ETHANOLIC EXTRACT FROM THE PSEUDOBULBS OF Grammatophyllum speciosum Blume

 

(Validasi Kaedah FT-KCPT bagi Kuantifikasi Gastrodin di dalam Ekstrak Etanolik dari Pseudobulbs Grammatophyllum speciosum Blume)

 

Verisa Chowjarean1, 2, Apirada Sucontphunt2, Saowapak Vchirawongkwin3, Tossaton Charoonratana4,

Thanapat Songsak4, Saraporn Harikarnpakdee2, 5, Parkpoom Tengamnuay6*

 

1Department of Pharmaceutical Technology Faculty of Pharmacy

2Cosmeceutical Research, Development and Testing Center, Faculty of Pharmacy

3Department of Pharmaceutical Chemistry, Faculty of Pharmacy

 4Department of Pharmacognosy, Faculty of Pharmacy

5Department of Industrial Pharmacy, Faculty of Pharmacy

Rangsit University, Pathumthani 12000, Thailand

6Department of Pharmaceutics and Industrial Pharmacy, Faculty of Pharmaceutical Sciences,

Chulalongkorn University, Bangkok 10330, Thailand

 

*Corresponding author:  parkpoom.t@chula.ac.th

 

 

Received: 7 September 2017; Accepted: 7 February 2018

 

 

Abstract

A reversed phase-high performance liquid chromatography-diode array detector (RP-HPLC-DAD) method was developed to determine the amount of gastrodin, the major active component of Grammatophyllum speciosum pseudobulbs.  A previous study suggested that the ethanolic extract of G. speciosum had a potential to encourage stemness of keratinocytes.  Therefore, in order to determine the quality of G. speciosum ethanolic extract, the quantitative analysis of gastrodin should be validated.  The optimized RP-HPLC condition was achieved within 18 min, using a column, Inertsil ODS-3 (4.6 x 150 mm, 5 µm).  The mobile phase consisted of a mixture of water: acetonitrile with a gradient from 1:99 to 100:0 in 14 minutes and keep constant at 1:99 for 4 minutes.  The flow rate was 1.7 mL/min with the monitored UV wavelength of 220 nm.  Gastrodin was eluted at the retention time of 6.5 minutes.  The calibration curve of gastrodin at the concentration of 10 - 100 µg/mL showed good linearity (r2 = 0.999).  In addition, the intraday and interday precisions of gastrodin were 1.05 and 0.38%, respectively.  The percentage recovery was within 99.3 - 101.5%.  Average gastrodin contents in 3 samples were 55.52 - 58.12 mg/g.  The results indicate that this method can be utilized to control the quality of G. speciosum extract.

 

Keywords:  gastrodin, Grammatophyllum speciosum, reversed-phase high performance liquid chromatography, quantification

 

Abstrak

Kaedah fasa terbalik-kromatografi cecair berprestasi tinggi-pengesan susunan diod (FT-KPCT-PSD) telah dibangunkan untuk menentukan jumlah kandungan gastrodin, komponen aktif utama bagi pseudobulbs Grammatophyllum speciosum. Kajian sebelumnya mencadangkan ekstrak etanolik terhadap G. speciosum mempunyai potensi menguatkan stem keratinocytes. Oleh demikian, bagi menentukan kualiti ekstrak etanolik G. speciosum, analisis kuantitatif terhadap gastrodin perlu ditentusahkan. Keadaan optimum FT-KCPT telah dicapai pada minit ke 18, mengunakan turus Inertsil ODS-3 (4.6 x 150 mm, 5 µm). Fasa bergerak mengandungi campuran air:asetonitril dengan cerunan dari 1:99 hingga 100:0 bagi tempoh 14 minit dan kekal malar pada 1:99 selama 4 minit. Kadar aliran ialah 1.7 mL/min dengan pemerhatian panjang gelombang UV pada 220 nm. Gastrodin terelusi pada masa tahanan ialah minit ke 6.5. Lengkung kalibrasi gastrodin pada aras kepekatan 10 - 100 µg/mL telah menunjukkan kelinearan yang baik (r2 = 0.999). Kejituan intra-hari dan inter-hari telah dicapai masing-masing pada 1.05 dan 0.38%. Peratusan perolehan semula berada di dalam julat 99.3 - 101.5%. Purata kandungan gastrodin  di dalam 3 sampel adalah 55.52 - 58.12 mg/g. Keputusan ini menunjukkan kaedah boleh digunapakai untuk mengawal kualiti G. speciosum yang diekstrak.

 

Kata kunci:      gastrodin, Grammatophyllum speciosum, fasa terbalik kromatografi cecair berprestasi tinggi, kuantifikasi

 

References

1.       Nontachaiyapoom, S., Sasirat, S. and Manoch, L. (2011). Symbiotic seed germination of Grammatophyllum speciosum Blume and Dendrobium draconis Rchb. f., native orchids of Thailand. Scienta Horticultuae, 130(1): 303 - 308.

2.       Sahakitpichan, P., Mahidol, C., Disadee, W., Chimnoi, N., Ruchirawat, S. and Kanchanapoom, T. (2013). Glucopyranosyloxybenzyl derivatives of (R)-2-benzylmalic acid and (R)-eucomic acid, and an aromatic glucoside from the pseudobulbs of Grammatophyllum speciosum. Tetrahedron, 69(3): 1031 - 1037.

3.       Uawonggul, N., Chaveerach, A., Thammasirirak, S., Arkaravichien, T., Chuachan, C. and Daduang, S. (2006). Screening of plants acting against Heterometrus laoticus scorpion venom activity on fibroblast cell lysis. Journal of Ethnopharmacology, 103 (2): 201 - 207.

4.       Kumar, H., Kim, I. S., More, S. V., Kim, B. W., Bahk, Y. Y. and Choi, D. K. (2013). Gastrodin protects apoptotic dopaminergic neurons in a toxin-induced Parkinson’s disease model. Journal of Evidence-Based Complementary and Alternative Medicine, 2013: 1 - 14.

5.       Hsieh, M. T., Wu, C. R. and Chen, C. F. (1997). Gastrodin and p-hydroxybenzyl alcohol facilitate memory consolidation and retrieval, but not acquisition, on the passive avoidance task in rats. Journal of Ethnopharmacology, 56(1): 45 - 54.

6.       Wu, C. R., Hsieh, M. T., Huang, S. C., Peng, W. H., Chang, Y. S. and Chen, C. F. (1996). Effects of Gastrodia elata and its active constituents on scopolamine-induced amnesia in rats. Planta Medica, 62 (4): 317 - 321.

7.       An, S. J., Park, S. K., Hwang, I. K., Choi, S. Y., Kim, S. K., Kwon, O. S., Jung, S. J., Baek, N. I., Lee, H. Y. and Won, M. H. (2003). Gastrodin decreases immunoreactivities of γ-aminobutyric acid shunt enzymes in the hippocampus of seizure-sensitive gerbils. Journal of Neuroscience Research, 71(4): 534 - 543.

8.       Kim, H. J., Moon, K. D., Lee, D. S. and Lee, S. H. (2003). Ethyl ether fraction of Gastrodia elata Blume protects amyloid β peptide-induced cell death. Journal of Ethnopharmacology, 84(1): 95 - 98.

9.       Hu, Y., Li, C. and Shen, W. (2014). Gastrodin alleviates memory deficits and reduces neuropathology in a mouse model of Alzheimer's disease. Neuropathology, 34(4): 370 - 377.

10.    Wang, X. L., Xing, G. H., Hong, B., Li, X. M., Zou, Y., Zhang, X. J. and Dong, M. X. (2014). Gastrodin prevents motor deficits and oxidative stress in the MPTP mouse model of Parkinson's disease: Involvement of ERK1/2–Nrf2 signaling pathway. Life Sciences, 114(2): 77 - 85.

11.    Huang, Q., Shi, J., Gao, B., Zhang, H. Y., Fan, J., Li, X. J., Fan, J. Z., Han, Y. H., Zhang, J. K. and Yang, L. (2015). Gastrodin: an ancient Chinese herbal medicine as a source for anti-osteoporosis agents via reducing reactive oxygen species. Bone, 73: 132 - 144.

12.    Chowjarean, V., Nimmannit, U., Chaotham, C., Eaknai, W., Sucontphunt, A., Plaimee, P. P., Tengamnuay, P. and Chanvorachote, P. (2017). Grammatophyllum speciosum extract potentiates stemness in keratinocyte cells. Chiang Mai Journal of Science, 45 (1): 237 - 248.

13.    Chen, W. C., Lai, Y. S., Lu, K. H., Lin, S. H., Liao, L. Y., Ho, C. T. and Sheen, L. Y. (2015). Method development and validation for the high-performance liquid chromatography assay of gastrodin in water extracts from different sources of Gastrodia elata Blume. Journal of Food and Drug Analysis, 23 (4): 803 - 810.

14.    Jia, Y., L,i X., Xie, H., Shen, J., Luo, J., Wang, J., Wang, K. D., Liu, Q. and Kong, L. (2014). Analysis and pharmacokinetics studies of gastrodin and p-hydroxybenzyl alcohol in dogs using ultra fast liquid chromatography–tandem mass spectrometry method. Journal of Pharmaceutical and Biomedical Analysis, 99: 83 - 88.

15.    Lee, J-G., Moon, S-O., Kim, S-Y., Yang, E-J., Min, J-S., An, J-H., Choi, E-A., Liu, K-H., Park, E. J. and Lee, H-D. (2015). Rapid HPLC determination of gastrodin in Gastrodiae Rhizoma. Applied Biological Chemistry, 58 (3): 409 - 413.

16.    AOAC Official Methods of Analysis (2013). Appendix K: Guideline for dietary supplements and botanicals.   http://www.eoma.aoac.org/app_k.pdf. [Accessed online 29 June 2017].

17.    Shrivastava, A. and Gupta, V. B. (2011). Methods for the determination of limit of detection and limit of quantitation of the analytical methods. Chronicles of Young Scientists, 2(1): 21 - 21.

 




Previous                    Content                    Next