subject: Aconitum Alkaloids In The Comparative Study Of Extraction - Aconitum - Pharmaceutical Industries [print this page] Pharmacognosy in the DDA and processed products, alkaloids is efficacy component is also toxic components [1], and stability is not good, is easy to degrade [2], so alkaloid extraction method is in its composition analysis important. Extraction of the alkaloids present there are many [3 ~ 5], however, various methods diester alkaloids Comparison of extraction efficiency there is no report. Chinese medicine used in the clinic DDA Traditional Chinese medicine The processed products as aconite, monkshood commercially available processed products of slices under different processing methods into salt Aconite, Monkshood black and white film attached, light aconite, monkshood cooked attached sheets and guns, etc. [6], but a different dish approach will definitely lead to different alkaloid content, the use should be different. This article compares the seven kinds of extraction methods on Aconite in three kinds of double-ester alkaloids extraction efficiency, and determination of aconitine in crude drugs and their processed products alkaloids to accurate determination of aconite alkaloids, To ensure the clinical application of these Chinese medicines more safe and reasonable to provide a reliable analysis.
1.2 reagent aconitine aconitine and hypaconitine reference standard was purchased from Pharmaceutical and Biological Products. Health monkshood, Aconitum purchased from Sichuan Jiangyou Health Kusnezoffii, black attached piece, attached to pieces of white, salt, aconite, monkshood light purchased from Jilin Province, Changchun City, Jilin deep pharmacies, by Changchun Medicine University Professor Wang Shumin identification are authentic. Chromatography of pure methanol and water as deionized water, ether, dichloromethane and other reagents were of analytical grade.
2 Methods and Results 2.1 Methodology test 2.1.1 Chromatographic conditions AgilentextendedC18 column (150mm 4.6mm, 5 m); mobile phase was methanol: 0.1% triethylamine (65:35); flow rate of 0.6ml? Min-1; detection wavelength 230nm; The column temperature was 30 ; sample through 0.45 m filter membrane, the filtrate sample 10 l.
2.1.2 standard curve precision weighed in aconitine aconitine aconitine and the second reference standard amount placed in flask, adding methylene chloride solution and set the volume, was the concentration of 1.08mg? Ml- 1 mesaconitine, 1.12mg? ml-1 of aconitine and 1.22mg? ml-1 of the sub-standard stock solution of aconitine. Control solution for each sample were 2,4,6,8,10 l, high performance liquid chromatography. In a concentration ( g? Ml-1) on the peak area was calculated regression curve. Be in the standard curve equation aconitine Y = 20672.0X-58377.5, r = 0.9995; aconitine standard curve equation Y = 18114.2X-28544.2, r = 0.9999; hypaconitine standard curve equation Y = 186632X -13386.7, r = 0.9993. Description aconitine in 5.6 ~ 112 g? Ml-1, in aconitine in 5.4 ~ 108 g? Ml-1, hypaconitine in the 6.1 ~ 122 g? Ml-1 concentration showed good linearity within.
2.1.3 repeatability of the same powder by Optimum Health Aconitum extraction method is to extract six copies of the solution, HPLC determination, record aconitine aconitine, hypaconitine peak area. Aconitine aconitine and times in Aconitine RSD values were 4.32%, 3.57% and 4.84%. Description method had good reproducibility.
2.1.4 recovery experiments of known precision weighed powder content of 18 students Aconitum, each 0.2g. 6 copies of the first one was added 0.05mg of aconitine, the first two 0.4mg were added 6 copies of aconitine, the first three 6 were hypaconitine were added to 0.2mg, with the calculation method is the content of aconitine The average recovery was 91.2%, RSD 2.84%; the average recovery of aconitine 90.4%, RSD = 3.12%; hypaconitine average recovery was 92.8%, RSD value of 3.10. Show that the recovery was good.
2.1.5 stability test sample solution the same health Aconitum powder stored in -80 refrigerator, the refrigerator an hour after the first 4,8,12,18,24 determination, calculation of concentration. 24h in aconitine aconitine and the RSD value of Aconitine were 5.21%, 4.35% and 6.24%. Show that the extract at -80 refrigerator stable within 24h.
2.2 Optimization of Extraction
2.2.1 Aqueous chloroform extraction method called in with a plug of Health Aconitum 2.0g conical flask, add 30ml distilled water, shaking after 6h, transferred to 50ml centrifuge tubes, 3000r? Min-1 centrifuge 20min, take supernatant, precipitate washed two times the combined supernatant was adjusted with 10% ammonia pH 10 were added 15,5,5 ml chloroform extract 3 times, combined chloroform layer, evaporation to dryness. Plus methanol solution, set the volume to 25ml.
