Validated high-performance thin-layer chromatography method for steviol glycosides in Stevia rebaudiana.
A high-performance thin-layer chromatographic (HPTLC) method was developed and validated as per ICH (International Conferences on Harmonization) guidelines for simultaneous quantification of three steviol glycosides, i.e. steviolbioside, stevioside and rebaudioside-A in Stevia rebaudiana leaves. For achieving good separation, mobile phase of ethyl acetate-ethanol-water (80:20:12, v/v/v) on pre-coated silica gel 60 F254 HPTLC plates were used. The densitometric quantification of steviol glycosides was carried out at lambda=510 nm in reflection-absorption mode after spraying with acetic anhydride:sulphuric acid:ethanol reagent. The calibration curves were linear in the range of 160-960 ng/spot for steviolbioside, 1-6 microg/spot for stevioside and 0.5-3 microg/spot for rebaudioside-A with good correlation coefficients (0.998-0.999). The method was found to be reproducible for quantitative analysis of steviol glycosides in S. rebaudiana leaves collected from ten different locations and will serve as a quality control indicator to monitor the commercial production of stevioside and its allied molecules during different stages of its processing.
Jaitak
Vikas
V
Natural Plant Products Division, Institute of Himalayan Bioresource Technology (CSIR), Palampur 176061, H.P., India.
Gupta
A P
AP
Kaul
V K
VK
Ahuja
P S
PS
D002138:Calibration; D002855:Chromatography, Thin Layer / Q000379:methods*; D045786:Diterpenes, Kaurane / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D005960:Glucosides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D006027:Glycosides / Q000032:analysis* / Q000737:chemistry / Q000302:isolation & purification; D015394:Molecular Structure; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D036625:Stevia / Q000737:chemistry*
D045786:Diterpenes, Kaurane; D005960:Glucosides; D006027:Glycosides; C074471:steviolbioside; C012043:stevioside; C012042:steviol