Identification of β‐sitosterol, stigmasterol and ergosterin in A. roxburghii using supercritical fluid extraction followed by liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry

Abstract

beta-Sitosterol and stigmasterol are the most common phytosterols in traditional Chinese medicine. They have been proved to have many important bioactivities. To the best of our knowledge, this is the first report that beta-sitosterol, stigmasterol and ergosterol coexisting in A. roxburghii herbs can be simultaneously extracted by a supercritical fluid extraction (SFE) procedure; then a simple high-performance liquid chromatography/atmospheric pressure chemical ionization ion trap mass spectrometry (HPLC/APCI/MS) method was developed for simultaneous identification and determination of these three compounds. The ion trap MS/MS detector was equipped with an atmospheric pressure chemical ionization source operating in the positive ion mode, APCI(+). The linear responses were obtained in the concentration range of 0.50-150 microg/mL (r = 0.9999) for ergosterol, 5-400 microg/mL (r = 0.9999) for stigmasterol, and 10-2000 microg/mL (r = 0.9998) for beta-sitosterol. An orthogonal L(9) (3(3)) test design was employed for optimization of the SFE process. Under the optimized conditions, i.e. pressure of 25 mPa, temperature of 45 degrees C and ethanol as modifier, the concentrations of sterols in the extract were found to be 2.89% (g/g) for beta-sitosterol, 3.56% (g/g) for stigmasterol and 2.96% (g/g) for ergosterin. The SFE method was also compared with a previously developed Soxhlet extraction. The SFE method produced higher yields of sterols than that of the Soxhlet extraction. The proposed method has been successfully used for identification and quantitation of beta-sitosterol, stigmasterol and ergosterin in a real A. roxburghii sample.

Identification and quantitation of auxins in plants by liquid chromatography/electrospray ionization ion trap mass spectrometry

“…The LC/ITMS system consists of an Esquire 3000 ion trap LC‐MS n system (Bruker Daltonik GmbH, Germany) and an 1100 Series LC system (Agilent, USA), which includes a quaternary pump, a diode‐array detector and a ChemStation system 31. Chromatographic separation was performed with an Agilent Eclipse XDB‐C18 column (5 µm, 4.6 × 150 mm) maintained at 25°C.…”

N‐alkylpyridinium quaternization for assisting electrospray ionization of sterols in oil by quadrupole‐time of flight mass spectrometry

“…[17][18][19][20][21][22][23] Zoosterols including cholesterol, dehydrocholesterol and dihydrocholesterol are usually minor components of vegetable oil, thus zoosterols in the oil show that the cooking oil could be illegally recycled from restaurants and other animal origins. The GC/MS combined with silylation [24,25] and atmospheric-pressure chemical ionization-MS [26][27][28][29][30][31] have been developed for the analysis of sterols in edible oil. However, these methods need tedious sample purification steps such as thin-layer chromatography, solid phase extraction and supercritical fluid extraction to remove other components and impurities due to the complexity of oil.…”

Simultaneous determination of nucleosides, myriocin, and carbohydrates in Cordyceps by HPLC coupled with diode array detection and evaporative light scattering detection

“…Several methods, including HPLC-DAD (diode array detector) [4,5], CE [6], CEC [7], LC-MS [8], ultra performance liquid chromatography [9], have been used for nucleosides analysis in Cordyceps. Sterols, especially ergosterol, have also been determined using HPLC-DAD [5], LC-MS [10] and GC-MS [11]. However, a few reports [12][13][14] focus on the carbohydrates such as mannitol, a characteristic component in Cordyceps, which have diuretic, decrease neutrophil apoptosis and antioxidant activities [15][16][17].…”