Crystallization-induced dynamic resolution R-epimer from 25-OCH3-PPD epimeric mixture.
25-OCH3-PPD is a promising antitumor dammarane sapogenin isolated from the total saponin-hydrolyzed extract of Panax ginseng berry and Panax notoginseng leaves. 20(R)-25-OCH3-PPD was more potent as an anti-cancer agent than 20(S)-25-OCH3-PPD and epimeric mixture of 25-OCH3-PPD. This paper describes the rapid separation process of the R-epimer of 25-OCH3-PPD from its epimeric mixture by crystallization-induced dynamic resolution (CIDR). The optimized CIDR process was based on single factor analysis and nine well-planned orthogonal design experiments (OA9 matrix). A rapid and sensitive reverse phase high-performance liquid chromatographic (HPLC) method with evaporative light-scattering detector (ELSD) was developed and validated for the quantitation of 25-OCH3-PPD epimeric mixture and crystalline product. Separation and quantitation were achieved with a silica column using a mobile phase consisting of methanol and water (87:13, v/v) at a flow rate of 1.0mL/min. The ELSD detection was performed at 50°C and 3L/min. Under conditions involving 3mL of 95% ethanol, 8% HCl, and a hermetically sealed environment for 72h, the maximum production of 25(R)-OCH3-PPD was achieved with a chemical purity of 97% and a total yield of 87% through the CIDR process. The 25(R)-OCH3-PPD was nearly completely separated from the 220mg 25-OCH3-PPD epimeric mixture. Overall, a simple and steady small-batch purification process for the large-scale production of 25(R)-OCH3-PPD from 25-OCH3-PPD epimeric mixture was developed.
School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China.
Sainan
S
Zhang
Key Laboratory of Natural Active Pharmaceutical of Jiangxi Province, Yichun University, Yichun 336000, China.
Yun
Y
Tang
Key Laboratory of Structure-based Drug Design and Discovery of Ministry of Education, Shenyang Pharmaceutical University, Shenyang 110016, China.
Jiaqing
J
Cao
School of Life Science and Biological Pharmacy, Shenyang Pharmaceutical University, Shenyang 110016, China.
Chen
C
Zhao
School of Traditional Chinese Materia Medica, Shenyang Pharmaceutical University, Shenyang 110016, China; Key Laboratory of Structure-based Drug Design and Discovery of Ministry of Education, Shenyang Pharmaceutical University, Shenyang 110016, China. Electronic address: zyq4885@126.com.
Yuqing
Y
Zhao
D000704:Analysis of Variance; D000970:Antineoplastic Agents / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003460:Crystallization / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D007536:Isomerism; D008027:Light; D016014:Linear Models; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D014315:Triterpenes / Q000032:analysis* / Q000737:chemistry* / Q000302:isolation & purification / Q000494:pharmacology; D000097669:Dammaranes
D000970:Antineoplastic Agents; D014315:Triterpenes; C517965:25-methoxydammarane-3,12,20-triol; D000097669:Dammaranes
25-OCH(3)-PPD epimeric mixture; Anticancer activity; Crystallization-induced dynamic resolution; HPLC-ELSD; Separation and quantitation