STABILITY INDICATING HPLC METHODS FOR CYCLODEXTRIN
DERIVATIVES
Gábor Varga1, Krisztina Ludányi3, Julianna Szemán2, Imre Klebovich3, Lajos Szente2
1
ChiroQuest Chiral Technologies Development Ltd., Budapest, Hungary
2 CycloLab Cyclodextrin R&D Laboratory Ltd., Budapest, Hungary, e-mail: szeman.j@cyclolab.hu
3 Semmelweis 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
ADC1 A, ADC1 (F:\DATA\CTZ0509\HPBHCL4O.D)
Methylated maltooligomers
50
100
45
80
600
800
1000
2.5
m/z
5
7.5
10
12.5
15
17.5
[V]
1342.4
DS: 12
1320
1340
1360
1380
1400
*MSD1 SPC, time=20.516:26.535 of D:\DOC\MS\CD_HCL.D
1407.4
1464.4
1465.2
1348.4
1466.2
1349.2
1302.4
1350.2
1246.4
1290.2
1245.4
1140.4
1141.4
1142.4
1186.4
1083.4
1084.4
1024.4
978.6
979.6
1006.4
1025.4
922.6
863.6
844.6
864.6
760.6
786.6
804.6
1244.4
921.6
759.6
701.6
703.8
624.6
643.6
642.6
682.6
596.8
539.8
481.8
422.8
1580.4
1581.2
1638.2
1524.2
1639.2
1582.2
1696.2
1640.2
1491.0
[min.]
1400
DS: 7
1600
1650
1700
m/z
API-ES, Pos, Scan, Frag: 150
API-ES, Pos, Scan, Frag: 150
API-ES, Pos, Scan, Frag: 150
API-ES, Pos, Scan, Frag: 150
API-ES, Pos, Scan, Frag: 150
DS: 4
Max: 5222
300000
250000
1505.2
1446.4
1550
1620.4
80
1500
Extracted ion chromatogram
1563.2
D S: 15
1450
MSD1 1407, EIC=1405:1409 (D:\DOC\MS\CD_HCL.D)
MSD1 1465, EIC=1463:1467 (D:\DOC\MS\CD_HCL.D)
MSD1 1523, EIC=1522:1525 (D:\DOC\MS\CD_HCL.D)
MSD1 1581, EIC=1579:1583 (D:\DOC\MS\CD_HCL.D)
MSD1 1639, EIC=1637:1641 (D:\DOC\MS\CD_HCL.D)
1504.4
D S: 12
1366.4
30
API-ES, Pos, Scan, Frag: 150
600000
DS: 8
200000
DS: 4
100000
1736.2
1680.2
1622.2
1591.8
1534.0
1678.2
1564.2
1506.2
1472.4
1482.4
1424.4
1448.2
1389.2
1366.4
1342.4
1320.4
1308.4
1331.2
1272.4
150000
50000
0
5
10
15
20
25
m in
0
1200
1300
1400
1500
1600
1700
m/z
5
CONCLUSIONS
15
20
25
Extracted ion chromatogram
 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
DS3
500000
400000
300000
200000
BCD
 The acidic degradation of CD derivatives resulted in substituted linear dextrins,
which show the same complexity as the parent cyclodextrins
DS1
600000
100000
0
2.5
5
7.5
REFERENCES
ACKNOWLEDGEMENT
[1]
[2]
[3]
[4]
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).
J. Szemán, K. Csabai, K. Kékesi, l. Szente, G. Varga; J. Chromatography A, 1116, 76-82 (2006)
S. Kawakishi, A. Satake, T. Komiya, M. Namiki; Starch/Stärke 25 203-206 (1983)
K. Uchida, S. Kawakishi; Agricult. Biol. Chem. 50(2) 54-57 (1986)
É. Fenyvesi, K. László; Cyclodextrin News 15(11) 203-206 (2001)
10
DS4
1400 m/z
DS2
1380
1180.4
1192.4
1204.4
1214.4
1380.4
20
1382.4
1375.4
1368.4
D S: 9
1330.4
40
200000
1447.2
1388.4
400000
1250.4
1262.4
1273.2
1284.4
1367.4
1621.2
60
1369.4
1361.4
1362.4
1363.4
25
HPBCD components DS: 1-10
1353.4
1354.4
1355.4
1349.4
1348.4
1347.4
1339.4
1340.4
1341.4
20
Time
m/z
A P I-E S , P o s, S c a n, F ra g : 1 5 0
A P I-E S , P o s, S c a n, F ra g : 1 5 0
A P I-E S , P o s, S c a n, F ra g : 1 5 0
DS: 14
1333.4
1334.4
1335.4
376.8
1523.4
15
100
1360
1465.2
1466.2
10
1562.4
M S D 1 1 3 0 1 , E IC = 1 2 9 9 :1 3 0 3 (D :\D O C \M S \R _ H C L .D )
M S D 1 1 3 4 3 , E IC = 1 3 4 1 :1 3 4 5 (D :\D O C \M S \R _ H C L .D )
M S D 1 1 3 8 5 , E IC = 1 3 8 3 :1 3 8 7 (D :\D O C \M S \R _ H C L .D )
1352.4
1338.4
1300
Extracted ion chromatogram
1324.4
1325.4
1326.4
1327.4
1319.4
1321.4
1312.4
1313.4
1305.4
5
Max: 13261
1340
1461.8
0
0
1280
1310.4
1320
283.8
301.8
1418.4
0
[ min. ]
800000
1311.4
1296.4
1297.4
1298.4
1284.4
1269.4
1283.4
1268.4
1282.4
1300
Max: 12226
1406.4
1398.4
1399.4
1386.4
1387.4
1379.4
1373.4
1365.4
1367.4
1359.4
1351.4
1353.4
20
0
1280
DS: 4
20
0
1260
m/z
1464.4
1385.4
1372.4
40
30
*MSD1 SPC, time=18.815:25.791 of D:\DOC\MS\R_HCL.D API-ES, Pos, Scan, Frag: 150
20
244.8
60
0
40
212.8
DS: 6
Voltage
1384.4
1371.4
1357.4
1358.4
1344.4
1345.4
1337.4
1339.4
1330.4
1331.4
1311.4
0
1325.4
1301.4
1316.4
1300.4
20
80
1400
API-ES, Pos, Scan, Frag: 150
40
1315.4
DS: 9
50
100
1200
DS: 6
1317.4
40
RAMEB components DS: 8-16
1000
60
1343.4
1329.4
V o lta g e
1314.4
DS: 15
60
Time
800
80
100
25
600
100
1370.4
1356.4
MS detection
80
80
20
400
Linear, hydroxypropylated maltoheptaoses
100
Max: 18633
60
200
*MSD1 SPC, time=13.240:19.401 of D:\DOC\MS\CD_HCL.D
1328.4
100
15
184.8
min
*MSD1 SPC, time=12.067:18.429 of D:\DOC\MS\R_HCL.D API-ES, Pos, Scan, Frag: 150
10
242.8
203.0
400
Linear, methylated maltoheptaoses
5
109.0
25
141.0
949.4 948.4
963.6
976.4
20
1522.4
[V ]
0
714.8
480.8
318.8
962.6
30
0
200
150
862.6
35
0
MS detection
1082.4
60
40
758.6
773.6
788.6
802.6
435.8
625.8
582.8
597.8
406.8
323.8
283.8
301.8
216.8
230.8
244.8
117.0
156.8
20
624.8
420.8
434.8
40
5GL
7-8-9Me
772.6
787.6
2GL
3-4-5Me
methylated
glucose
743.8
596.8
60
538.8
Degradation
products
40
1408.2
80
200
Max: 5060
920.6
4GL
5-6-7-8Me
Max: 5210
700.6
3GL
4-5-6-7Me
*MSD1 SPC, time=4.391:13.497 of D:\DOC\MS\CD_HCL.D API-ES, Pos, Scan, Frag: 150
1406.4
786.6
100
HPBCD
ELSD detection
API-ES, Pos, Scan, Frag: 150
610.8
*MSD1 SPC, time=2.003:12.525 of D:\DOC\MS\R_HCL.D
758.6
ELSD detection
Hydroxypropylated maltooligomers
mAu
10
12.5
15
17.5
min
min