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
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