Improvement method of fast and isocratic RP-HPLC analysis of major diterpene glycoside from Stevia rebaudiana leaves

Abstract

Hemoglobin level after ingestion of repeated dose of iron shortly after and 2 hours after meal for 12 weeks in second & third trimester pregnant women with iron deficiency anemia AIP Conference Proceedings 1755, 160004 (2016) Abstract. Stevioside and rebaudioside A are major diterpene glycoside from Stevia rebaudiana. These compounds have similar structure and make both compounds have close polarity. The aim of this study was to improve isocratic RP-HPLC analysis to achieve a complete separation of stevioside and rebaudioside A in baseline peak. The method improved was conducted on Eurosphere C-18 (250 × 4.6 mm, 5 µm) as the stationary phase. Mobile phase was made by a mixture of water-methanol (90: 10, v / v, pH = 3.0); acetonitrile, and trifluoroacetic acid (TFA) in the ratio 65: 35: 0.01 (v/v/v). Flow rate of mobile phase was 0.6 mL/min. The detection was performed in UV-210 nm. The improved method has complete separation by R > 2.00 in baseline peak. The linearity of the standard curve was obtained with a correlation coefficient of 0.9998 for rebaudioside A and 0.9994 for stevioside. The percent recovery was determined by standard addition method with a mean recovery of 100.82% for rebaudioside A and 101.04% for stevioside. The relative standard deviation (RSD) of intraday precision varies from 0.34 -1.79% for rebaudioside A and 0.31 -1.81% for stevioside, and interday precision showed RSD 1.47% for rebaudioside A and 0.37% for stevioside. While the sensitivity reported in the limit of detection (LOD) value of method improvement was 1.00 µg/mL for both rebaudioside A and stevioside. The limit of quantitation (LOQ) of the method was 3.00 µg/mL for both respected compounds. The small changes of the mobile phase composition, the mobile phase is reused after 1 week, flow rate, and column temperature did not alter the method performance significantly. The reliable and robust of the developed and validated method was applied to both simple and complex matrices of samples including S. rebaudiana leaves for major diterpene glycoside analysis.

Pendidikan Keluarga: Perang Orang Tua terhadap Pembentukan Karakter Remaja Di Gereja Kristen Jawa Cilacap Utara

“…Perubahan sosial memberikan dampak terjadinya pergeseran nilainilai dalam masyarakat. Perubahan sosial menuntut manusia untuk berubah baik secara perlahan maupun cepat untuk mengikuti perkembangan era (Martono, 2016). Peranan era digitalisasi, baik dalam sosial media dan ekonomi, menempatkan perubahan sosial untuk mempengaruhi pertumbuhan moral anak remaja.…”

Daminozide enhances the vigor and steviol glycoside yield of stevia (Stevia rebaudiana Bert.) propagated in temporary immersion bioreactors

“…Accumulation of stevioside and rebaudioside A was analyzed using high-performance liquid chromatography (HPLC) based on method reported by Martono et al (2016). Sample for analysis was prepared by drying the leaves of stevia in the oven at 105° C for ve hours.…”

Determination of Stevioside and Rebaudioside A in <i>Stevia rebaudiana</i> Bertoni Leaves Using near Infrared Spectroscopy and Multivariate Data Analysis

“…Sample extraction conducted was referred to Martono et al [8] method. A 0.5 g of each dried powder S. rebaudiana leaves were weighed and extracted three times with 25 mL ethanol 60% (v/v) in every extraction using sonication extraction at 40 °C for 15 min every extraction cycle.…”

“…Method for HPLC analysis was explained in previous publications by Martono et al [8]. HPLC analysis was conducted on Eurosphere C-18 (250 × 4.6 mm i.d, 5μm) with guard column.…”

“…The polarity of Stevioside A is slightly higher than that of stevioside. Most of the methods reported based on HPLC method [1][2][6][7][8][9][10][11]. However, HPLC method is still costly, need a long time to analyze, high consume on solvents and unpracticed in sample handling.…”