STABILITY INDICATING HPLC METHODS FOR CYCLODEXTRIN DERIVATIVES

1. *MSD1 SPC, time=2.003:12.525 of D:\DOC\MS\R_HCL.D API-ES, Pos, Scan, Frag: 150 786.6 100 3GL 4-5-6-7Me 4GL 5-6-7-8Me 610.8 Max: 5210 80 2GL 3-4-5Me 5GL 7-8-9Me 60 596.8 methylated glucose 40 787.6 420.8 962.6 772.6 434.8 624.8 406.8 20 323.8 597.8 963.6 948.4 582.8 788.6 773.6 156.8 244.8 283.8 625.8 230.8 301.8 758.6 802.6 435.8 216.8 949.4 976.4 117.0 0 200 400 600 800 1000 m/z *MSD1 SPC, time=13.240:19.401 of D:\DOC\MS\CD_HCL.D API-ES, Pos, Scan, Frag: 150 1522.4 100 DS: 6 Max: 12226 1464.4 80 1580.4 1523.4 60 1465.2 1581.2 DS: 6 DS: 4 40 1638.2 1524.2 1466.2 1582.2 1639.2 1406.4 20 1640.2 1418.4 1696.2 1461.8 1491.0 0 1400 1450 1500 1550 1600 1650 1700 m/z *MSD1 SPC, time=20.516:26.535 of D:\DOC\MS\CD_HCL.D API-ES, Pos, Scan, Frag: 150 MSD1 1407, EIC=1405:1409 (D:\DOC\MS\CD_HCL.D) API-ES, Pos, Scan, Frag: 150 MSD1 1465, EIC=1463:1467 (D:\DOC\MS\CD_HCL.D) API-ES, Pos, Scan, Frag: 150 MSD1 1523, EIC=1522:1525 (D:\DOC\MS\CD_HCL.D) API-ES, Pos, Scan, Frag: 150 DS: 4 MSD1 1581, EIC=1579:1583 (D:\DOC\MS\CD_HCL.D) API-ES, Pos, Scan, Frag: 150 1562.4 MSD1 1639, EIC=1637:1641 (D:\DOC\MS\CD_HCL.D) API-ES, Pos, Scan, Frag: 150 100 DS: 7 DS: 14 Max: 5222 1504.4 300000 80 1563.2 250000 DS: 8 1620.4 1505.2 1446.4 200000 60 1621.2 DS: 4 150000 1447.2 40 1388.4 1564.2 1506.2 1330.4 100000 1622.2 1678.2 1389.2 1272.4 1448.2 1331.2 20 1366.4 1424.4 1214.4 1273.2 1308.4 1534.0 1204.4 1680.2 1591.8 1262.4 50000 1482.4 1250.4 1192.4 1736.2 1180.4 1284.4 1342.4 1320.4 1472.4 0 0 1200 1300 1400 1500 1600 1700 m/z 5 10 15 20 25 min STABILITY INDICATING HPLC METHODS FOR CYCLODEXTRIN DERIVATIVES Gábor Varga1, Krisztina Ludányi3, Julianna Szemán2, Imre Klebovich3, Lajos Szente2 1ChiroQuest Chiral Technologies Development Ltd., Budapest, Hungary 2 CycloLabCyclodextrin R&D Laboratory Ltd., Budapest, Hungary, e-mail: szeman.j@cyclolab.hu 3Semmelweis University, Faculty of Pharmacy, Department of Pharmaceutics, Hőgyes Endre u. 7, Budapest, H-1092, Hungary INTRODUCTION Forced degradation of CDs The characterisation of the isomer distribution and purity of cyclodextrin (CD) derivatives, their routine quality control and examination of their stability during storage are still a real problem. Using even the most sophisticated analytical methods the separation and identification of all components is far beyond the possibility. CD-Screen column designed for cyclodextrin analysis contains susbstituted phenyl groups bonded to silicagel stationary phase. This stationary phase suitable for fingerprint characterization of different CD derivatives, as well as, gives the possibility to follow their degradation [1]. CDs and CD derivatives are relatively stable substances, only a few articles can be found on their decomposition [2.3,4]. However, to follow the hydrolytical, oxidative or enzymatic decomposition of CDs and their derivatives can be interesting not only as research subject, but also from practical point of view. In this work our aim was to develop stability indicating HPLC methods for CD derivatives, to follow their degradation pathways by studying the structures of the degradation products. • Samples stored under stress conditions: • 2-(hydroxy)propyl-b-cyclodextrin (HPBCD) • Randomly methylated b-cyclodextrin (RAMEB) • Decomposition under stress conditions: • In 1 M hydrochloric acid solution  moderate decomposition • In 1 M sodium hydroxide solution  no decomposition • In 30% hydrogen peroxide solution  slight decomposition RESULTS AND DISCUSSION Acidic decomposition of RAMEB Acidic decomposition of HPBCD Methylated maltooligomers Hydroxypropylated maltooligomers ELSD detection ELSD detection Linear, methylated maltoheptaoses Linear, hydroxypropylated maltoheptaoses MS detection DS: 12 MS detection DS: 15 DS: 9 RAMEB components DS: 8-16 HPBCD components DS: 1-10 Extracted ion chromatogram Extracted ion chromatogram CONCLUSIONS Extracted ion chromatogram • The acidic degradation of CD derivatives resulted in substituted linear dextrins, which show the same complexity as the parent cyclodextrins • The first step of the acidic hydrolysis is the ring opening; the further fragmentation of the substituted maltoheptaoses is faster in case of HPBCD than in case of RAMEB • The obtained information provides the theoretical basis of the future development: development of a simple method using even RI or ELS detection for quantitation of the formed decomposition products of cyclodextrin derivatives REFERENCES ACKNOWLEDGEMENT [1] J. Szemán, K. Csabai, K. Kékesi, l. Szente, G. Varga; J. Chromatography A, 1116, 76-82 (2006) [2] S. Kawakishi, A. Satake, T. Komiya, M. Namiki; Starch/Stärke 25 203-206 (1983) [3]  K. Uchida, S. Kawakishi; Agricult. Biol. Chem. 50(2) 54-57 (1986) [4]  É. Fenyvesi, K. László; Cyclodextrin News 15(11) 203-206 (2001) The authors are grateful to Ms. Zs. Zachár and Ms. E. Erdei to their valuable technical assistance. The work was supplied by the National Research Fund (NKFP-1A-041/2004 and NKFP1-012/2005).