Quantitative study of ten main components in Shuangshenlong Capsule by HPLC coupled with wavelength switching and gradient elution method / 中草药

ABSTRACT

Objective: To establish HPLC coupled with wavelength switching and gradient elution method (HPLC-DVD) for simultaneous determination of ten main components (calycosin-7-glucoside, ruscogenin, amygdalin, ginsenoside Rb1, ginsenoside Re, ferulic acid, crocin I, salvianolic acid B, acetyl-11-keto-β-boswellic acid, and tanshinone IIA) in Shuangshenlong Capsule (SSLC). Methods: The chromatographic separation was achieved on Hypersil ODS C18 (150 mm × 4.0 mm, 3 μm) column with methanol- water (8∶2) T (A)-0.1% phosphoric acid solution (B) as mobile phases for gradient elution, at the flow rate of 0.6 mL/min; The detection wavelength was set at 260 nm for α-calycosin-7-glucoside, 280 nm for ruscogenin, 210 nm for amygdalin, 203 nm for ginsenoside Rb1 and ginsenoside Re, 320 nm for ferulic acid, 440 nm for crocin I, 286 nm for salvianolic acid B, 250 nm for acetyl-11-keto-β-boswellic acid, and 270 nm for tanshinone IIA. The volume of sample injection was 10 μL. Results:The ten active components were well separated and showed good linearity between mass concentration and peak area, such as calycosin-7-glucoside 3.88—69.86 mg/L (r = 0.999 2), ruscogenin 22.1—397.8 mg/L (r = 0.999 1), amygdalin 37.43—673.5 mg/L (r = 0.999 4), ginsenoside Rb1 45.15—812.72 mg/L (r = 0.999 6), ginsenoside Re 4.55—81.95 mg/L (r = 0.999 5), ferulic acid 3.06—55.15 mg/L (r = 0.999 4), crocin I 1.93—34.76 mg/L (r = 0.999 5), salvianolic acid B 15.68—282.15 mg/L (r = 0.999 6), acetyl-11-keto-β-boswellic acid 11.31—203.58 mg/L (r = 0.999 1), and tanshinone IIA 1.89—34.16 mg/L (r = 0.999 6). The precision was good, and RSD was not more than 1.27%. The repeatability was good, and RSD was not more than 1.28%. The stability was good in 8 h, and RSD was not more than 0.96%. The average recoveries and corresponding RSD values were 99.61% (1.21%), 100.11% (0.76%), 101.52% (0.62%), 101.22% (1.03%), 100.83% (1.14%), 98.94% (0.53%), 101.04% (1.09%), 100.05% (1.25%), 99.81% (0.68%), and 101.94% (1.31%), respectively. The contents of nine batches of calycosin-7-glucoside, ruscogenin, amygdalin, ginsenoside Rb1, ginsenoside Re, ferulic acid, crocin I, salvianolic acid B, acetyl-11-keto-β-boswellic acid, and tanshinone IIA were 0.142—0.158, 0.747—0.764, 1.578—1.619, 2.163—2.185, 0.235—0.251, 0.557—0.580, 0.105—0.122, 0.311—0.328, 0.605—0.624, 0.062—0.079 mg/capsule, respectively. Conclusion: HPLC coupled with wavelength switching and gradient elution method has been established for simultaneous determination of ten components in SSLC. The method is simple, quick, accurate, and it can be used for content determination and quality control of SSLC.