ABSTRACT
Objective::To establish an LC-MS/MS method for the determination of alcohol-soluble components and water-soluble components in Arnebiae Radix(L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid), for the purpose of quality control of the herb. Method::Chromatographic separation was carried out at 35 ℃ on Agilent ZORBAX SB C18 (4.6 mm×50 mm, 1.8 μm), with 0.1% formic acid acetonitrile (A)-0.1% formic acid solution (B) as the mobile phase for gradient elution (0-3 min, 5%-95%A; 3-7 min, 95%A; 7-7.01 min, 95%-5%A; 7.01-8.5 min, 5%A). The flow rate was 0.2 mL·min-1, and the injection volume was 5 μL. In a negative ionization mode, MRM scanning mode was adopted for quantification. Result::The calibration curves of L-shikonin, β, β′-dimethylacrylshikonin, β-acetoxy-isovalerylshikonin, lithospermic acid, caffeic acid, rosmarinic acid showed a good linearity, and the linear ranges of the above six compounds were 10.12-1 012 μg·L-1(r=0.998 1), 10.88-1 088 μg·L-1(r=0.991 2), 10.08-806.4 μg·L-1(r=0.997 6), 20.32-1 016 μg·L-1(r=0.996 6), 10.37-1 037 μg·L-1(r=0.999 6), 10.26-1 026 μg·L-1(r=0.997 8), respectively. The average recoveries of the analysts were 95.8%(RSD 3.2%), 103.5%(RSD 2.3%), 105.3%(RSD 2.1%), 96.1%(RSD 3.3%), 98.9%(RSD 2.7%), 100.8%(RSD 3.4%). The contents of six components in 13 batches of samples showed significant differences. Conclusion::The established method is feasible and simple, and provides a basis for comprehensive quality evaluation of Arnebiae Radix.
ABSTRACT
This present study is to develop an HPLC method for simultaneous determination of eight hydroxyl naphthoquinones, shikonin, β-hydroxy-isovalerylshikonin, acetylshikonin, β-acetoxy-isovalerylshikonin, deoxyshikonin, isobutyrylshikonin, β,β'-dimethylacrylshikonin and isovalerylshikonin. The eight constituents were measured on a Waters Xbridge C18 column (4.6 mm×250 mm，5 μm) with isocratic elution of acetonitrile-0.05% formic acid solution (70∶30) at a flow rate of 1.0 mL•min⁻¹. The detection wavelength was 275 nm and the column temperature was 30 ℃. The results of content determination indicated that significant differences of the eight compounds exist in every part of Arnebia euchroma，in which the highest part was the root bark, followed with the root, then the stem residues. The content of the xylem of root and aerial part was lower than the above parts. The results provided scientific basis for the medicinal parts of A. euchroma.
ABSTRACT
To develop a determination method forβ,β'-dimethylacrylalkannin and salvianolic acid B in Shangyangyu ointment by solid phase extraction coupled with HPLC. Methods:Solid phase extraction coupled with HPLC was used with a C18 (250 mm × 4. 6 mm, 5 μm) column, the mobile phase was acetonitrile-water-methanol-formic acid (700∶300∶30∶15) with the flow rate of 1. 0 ml·min-1. For β, β'-dimethylacrylalkannin, the detection wavelength was 275nm, and for salvianolic acid B, the detection wavelength was 286nm. Under the above conditions, the contents of main componentsβ,β'-dimethylacrylalkannin and salvianolic acid B in Shangyangyu ointment were determined. Results:The linear relationship was promising when the concentration ofβ,β'-dimethyl-acrylshikonin was within the range of 23. 780-118. 900μg·ml-1(r=0. 999 5) with the recovery of 99. 5% (RSD=1. 07%) and that of salvianolic acid B was within the range of 19.840-99.200 μg·ml-1(r=0.999 3) with the recovery of 98.2% (RSD=2.1%). Conclusion:The method is simple, accurate and fast with good separation, which can be used in the quality control of Shangyangyu ointment.
ABSTRACT
Objective To investigate the variation of the preparation time of Yuhong Ointment under conditions of accelerated test and long-term test;To provide the necessary data for the production and new medicine application, and establish the period of validity. Methods Referring to the current guiding principles of TCM stability test, the conditions of affecting factors in trial test and long-term test were decided:β,β'-dimethylacrylshikonin content was set as quantitative indicators, combined with the key items of stability test to evaluate the stability;its validity predictive value was deduced by using statistical methods. Results In the conditions of high temperature (30 ± 2 ℃, RH 45% ± 5%), high humidity (25 ± 2 ℃, RH 75% ± 5%), and hard light (4500 ± 500 Lx, 25 ± 2 ℃, RH 45%± 5%) for 10 days, the traits, appearance and content were in line with requirements. The validity of Yuhong Ointment under 25 ℃ conditions was 41.216 months. Conclusion Under current production conditions, the stability of Yuhong Ointment is good.
ABSTRACT
OBJECTIVE: To establish a new chromatography technique to, separate and purify naphthaquinone components from rhizoma of Arnebia euchroma(Royle). METHODS: Medium and low pressure preparative chromatography and Flash column were used to separate and purify the constituents, and their structures were elucidated by physicochemical properties, IR, MS, 1H-NMR and C-NMR. RESULTS: Six monocases were obtained, which were deoxyshikonin, shikonin, isobutylshikonin, acetylshikonin, β-ace-toxyisovalerylshikonin, and β, β-dimethylacrylshikonin. The purities of the 6 compounds analyzed by HPLC were 99.62%, 99.02%, 99.5%, 99.31%, 99.3% and 99.2%, respectively. CONCLUSION: A medium and low pressure preparative chromatography method has been established for the first time which can accurately and rapidly separate monocases from herbs.