Comparison of calcium oxalate crystals- raphides, and oxalic acid content in chamerion angustifolium (l.) Holub during vegetation

Abstract

INTRODUCTION Chamerion angustifolium (L.) Holub -willowherb is a flowering perennial plant of the Onagraceae family, which is very promising in medicine and pharmacy practice. Traditionally, willow herb is used in medicine to treat variety of ailments such as benign prostatic hyperplasia, mouth and skin ulcers, burns. One the most common diagnostic anatomical feature of the Chamerion angustifolium (L.) Holub herb is calcium oxalate crystals- raphides, that are formed from oxalic acid.AIM To compare calcium oxalate crystals- raphides, and oxalic acid content in Chamerion angustifolium (L.) Holub herb during different vegetation phases - intensive growing, bud, beginning of flowering, massive blooming, end of vegetation. METHODS Dried material was soften with the mixture of glycerol-water-ethanol 1:1:1. The cross cut of the stalks and petiole was made. The calculations of the raphides in nine units of 0,01mm2 in each section were made by using a microscope „OLYMPUS CX21“ connected to the digital camera“ BMS Microscope Camera 5.0 Mp“ and computer program. The calculation were repeated three times. For determing oxalic acid - 2,5g of grounded dried willow herb material (intensive growing, bud, beginning of flowering, massive blooming, end of vegetation) was transfered to 25mL volumetric flask and the ultrapure water was added to the scale. Oxalic acid was extracted with ultra-sonication at 50 °C for 30min. Then 8 mL extracts was centrifuged at 8000r/min for 15min. The supernatant soliution was filtered through 0,45 μm nylon membrane filter. The extracts were chromatographed on Waters e2695, column YMC- Pack ODS- A(5μm, C18, 250x4,6mm), pre - column YMC-Triart (5 μm ,C18,10x3mm) with 0,2% phosphoric acid as mobile phase, and flow rate was 0,5mL/min. The column temperature was 40 °C and chromatography was monitored by the UV/vis detector at 210nm. The results of the tests were statistically processed using Microsoft Excel spreadsheet software.[...].