ABSTRACT
Objective To establish the HPLC fingerprint and simultaneous method for the determination of 11 saponins in steamed Panax notoginseng from different origins. Methods Agilent Zorbax SB-C18 (250 mm × 4.6 mm, 5 μm) column was adopted, the mobile phase consisted of acetonitrile-water with gradient elution at the flow rate of 1.0 mL/min (elution program: 0-30 min, 20% acetonitrile; 30-60 min, 20%-45% acetonitrile; 60-88 min, 45%-75% acetonitrile; 88-98 min, 75% acetonitrile; 98-100 min, 75%-20% acetonitrile), and the detection wavelength was 203 nm. The establishment of steamed P. notoginseng HPLC fingerprint, and determination method of notoginsenoside R1, ginsenoside Rg1, Re, Rb1, 20S-Rh1, 20R-Rh1, Rd, Rk3, Rh4, 20S-Rg3, and 20R-Rg3 index components were studied methodologically. The content of 11 saponins in 10 batches was determined. Results The HPLC fingerprint was establish, and thirty common peaks were selected as characteristic peaks of steamed P. notoginseng. The similarities of different samples from 10 areas were 0.941, 0.938, 0.945, 0.951, 0.913, 0.909, 0.920, 0.928, 0.917, and 0.919. In quantitative analysis, eleven saponins were separated well and the average content was 5.274, 20.515, 2.838, 23.651, 3.476, 1.407, 5.239, 1.784, 1.580,0.904, and 0.294 mg/g, respectively. Additionally, all calibration curves showed good linear regression relationship, with correlation indexes of 0.999 7, 0.999 5, 0.999 5, 0.999 7, 0.999 7, 0.999 6, 0.999 7, 0.999 6, 0.999 7, 0.999 7, and 0.999 6; The average recoveries were 101.23%, 98.52%, 97.67%, 99.62%, 98.17%, 98.92%, 99.44%, 99.14%, 100.25%, 98.23%, and 96.89%, with RSDs of 1.35%, 1.58%, 2.44%, 1.05%,1.48%, 1.56%, 0.85%, 2.34%, 2.85%, 1.25%, and 1.08%. Conclusion This method is sensitive, accurate and reproducible. It can be used to provide a reference for the standard and evaluation of quality of steamed P. notoginseng.
ABSTRACT
OBJECTIVE: To establish an HPLC method for determining four kinds of rare saponins, ie, 20(S)-Rg3, 20(R)-Rg3, Rk1, and Rg5 in black ginseng. METHODS: COSMOSIL C18-PAQ (4.6 mm×250 mm, 5 μm) column was used and temperature was maintained at 30℃. Gradient elution was conducted using mobile phase consisting of acetonitrile and water at a flow rate of 1.0 mL·min-1. The detection wavelength was set at 203 nm. The injected sample volume was 10 μL. RESULTS: Good resolution was achieved for the four rare saponins in the ranges of 0.051-0.256 (r=0.999 9), 0.009-0.280(r=0.999 7), 0.051-0.303(r=0.999 9) and 0.093-0.279 mg·mL-1(r=0.999 9) for 20(S)-Rg3, 20(R)-Rg3, Rk1, and Rg5, respectively. The corresponding average recovery rates were 103.9%, 99.2%, 97.0%, and 100.6%, and the standard deviations were 0.71%, 0.73%, 1.97%, and 0.57%, respectively. CONCLUSION: The method is accurate, simple, reliable and reproducible for the determination of saponins in black ginseng. The determination result can be used as a reference for the rational medication, quality control, and further study of black ginseng.