Rapid quantification of constituents in St. John's wort extracts by NIR spectroscopy.
A quantitative near-infrared reflectance spectroscopy (NIRS) method was established for the determination of two major constituents (hyperforin and I3,II8-biapigenin) in St. John's wort extracts. Hyperforin was chosen due to the fact that it is found in a concentration range from 1 to 5%, a common one for NIRS determinations. I3,II8-Biapigenin on the other hand was selected as a constituent with very low concentrations (0.1-0.7%) but an extensive chromophore that allows very precise measurements in the ultraviolet (UV) and thus exact reference values that are vital for proper NIRS calibrations. Reference measurements were performed by reversed-phase high performance liquid chromatography (HPLC), determining the constituents' content in 35 pharmaceutical dry extracts of different origins. The reference method was validated according to the ICH guideline Q2B. Using partial-least squares (PLS) regression a multivariate calibration was done for the two ingredients each (PLS1). Satisfactory calibration statistics were obtained for hyperforin with a root mean square error of calibration (RMSEC) of 0.17 and a root mean square error of prediction (RMSEP) of 0.22 at a concentration range from 1 to 6% in the dry extracts. Due to the very low concentrations of I3,II8-biapigenin the accuracy of prediction is somewhat lower. However, it is possible to obtain very good results and reliable prediction by dividing the concentration range at 0.35%. The study emphasizes the potential of NIRS as a rapid and highly effective alternative method to conventional quantitative analysis of plant extracts.
Rager
I
I
Pharmaceutical Institute, Eberhard-Karls-Universitat Tuebingen, Auf der Morgenstelle 8, D-72076 Tuebingen, Germany.
Roos
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G
Schmidt
P C
PC
Kovar
K-A
KA
D047310:Apigenin; D044946:Biflavonoids; D001643:Bridged Bicyclo Compounds; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D005419:Flavonoids / Q000032:analysis; D020902:Hypericum / Q000737:chemistry*; D015999:Multivariate Analysis; D010696:Phloroglucinol / Q000031:analogs & derivatives; D010936:Plant Extracts / Q000032:analysis; D012015:Reference Standards; D013056:Spectrophotometry, Ultraviolet; D019265:Spectroscopy, Near-Infrared; D013729:Terpenes / Q000032:analysis
D044946:Biflavonoids; D001643:Bridged Bicyclo Compounds; D005419:Flavonoids; D010936:Plant Extracts; D013729:Terpenes; C446502:biapigenin; D047310:Apigenin; D010696:Phloroglucinol; C001654:hyperforin