Application of Response Surface Methodology for Optimisation of Simultaneous UHPLC-PDA Determination of Oleanolic and Ursolic Acids and Standardisation of Ericaceae Medicinal Plants

Abstract

Abstract:A fast and sensitive ultra-high performance liquid chromatography-photodiode array (UHPLC-PDA) method for simultaneous quantification of oleanolic acid (OA) and ursolic acid (UA) in plant materials was developed. A central composite design combined with a response surface methodology was utilized to establish optimal separation conditions. The final separation was accomplished on a Zorbax Eclipse XDB-C18 column (1.8 µm, 100 mm × 3 mm I.D., Agilent, Santa Clara, CA, USA) using a mixture 90:10 (v/v) of methanol and 1% (w/v) aqueous orthophosporic acid as a mobile phase at a flow rate of 0.44 mL/min and temperature of 18 • C. The analysis was completed in 6.2 min with satisfactory resolution of 1.5 between the target analytes. The developed method proved to be precise (relative standard deviations below 3.2%), accurate (recoveries in the range of 95.27%-98.60%), and sensitive (limits of detection (LODs) in the range of 0.047-0.051 mg/mL). The method was then successfully applied to evaluate OA and UA content in real samples of selected Ericaceae plant materials (leaves of Arctostaphylos uva ursi, Vaccinium myrtillus, Vaccinium vitis idaea, Gaultheria procumbens). The content of OA and UA in investigated samples varied in the range of 0.74-4.47 mg/g dry weight (dw) and 1.30-18.61 mg/g dw, respectively.

Metabolite Profiling of Eastern Teaberry (Gaultheria procumbens L.) Lipophilic Leaf Extracts with Hyaluronidase and Lipoxygenase Inhibitory Activity

“…The triterpene acids UA and OA , recommended as analytical markers of anti-inflammatory active wintergreen lipophilic extracts, were simultaneously determined by UHPLC-PDA in the leaf samples of G. procumbens harvested monthly across the full growing season. The validated assay protocol, optimized to assure the best recovery of both analytes from plant materials, was previously developed in our laboratory [ 45 ] for standardization of Ericaceae herbal medicines.…”

“…The triterpene acids content in other Ericaceae plants is generally lower. For instance, the leaves of lingonberry ( Vaccinium vitis-idaea ) contained 2.50–6.76 mg/g DW of UA and 0.74–2.15 mg/g DW of OA [ 45 , 47 , 48 ], while the content of both acids in the bilberry leaves ( Vaccinium myrtillus ) varied in the range of 0.75–1.30 mg/g DW for UA and 0.28–1.78 mg/g DW for OA [ 45 , 49 , 50 ]. Similarly, the sum of UA and OA in the leaves of Rhododendron adamsii reached 3.1 mg/g DW [ 51 ].…”

“…UHPLC-PDA analyses were carried out on an Agilent Technologies 1290 Infinity chromatograph (Agilent Technologies) according to the method of Owczarek et al [ 45 ]. The system was equipped with a photodiode array detector (PDA), thermostat, and autosampler.…”

Optimization, Validation and Application of HPLC-PDA Methods for Quantification of Triterpenoids in Vaccinium vitis-idaea L.

“…High waste in terms of reagents together with the multi-step prolonged analysis may be a limiting factor for routine analysis. Other studies have only focused on main triterpenoid acids [ 33 , 34 ]. As far as we know, currently, no validated, rapid, and easy-to-perform quantification method is available for the routine control of different groups of triterpenoids in lingonberries.…”

Development and Validation of a Fast and Sensitive Uhplc-Pda Method for the Quantification of Ursolic Acid in Poly(l-Lactic Acid) Nanocapsules

“…In this scenario, some analytical methods were previously described for the UA quantification, mainly in botanicals [18,19] and biological samples as plasma [20,21]. Concerning to the methods developed for drug delivery systems, these strategies were typically based on highperformance liquid chromatography [22][23][24].…”