Irinotecan Hydrochloride
(eye'' ri noe tee' kan hye'' droe klor' ide).
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C33H38N4O6·HCl·3H2O Anhydrous: 623.14 Trihydrate: 677.18
[1,4¢-Bipiperidine]-1¢-carboxylic acid, 4,11-diethyl-3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinolin-9-yl ester, monohydrochloride, trihydrate, (S)-;    
(+)-7-Ethyl-10-hydroxycamptothecine 10-[1,4¢-bipiperidine]-1¢-carboxylate, monohydrochloride, trihydrate     [136572-09-3].
DEFINITION
Irinotecan Hydrochloride contains NLT 98.0% and NMT 102.0% of C33H38N4O6·HCl, calculated on the anhydrous basis.
IDENTIFICATION
•  B. The retention time of the major peak of the Sample solution corresponds to the irinotecan (S-enantiomer) peak in the Identification solution, as obtained in the test for Limit of Irinotecan Hydrochloride Enantiomer.
•  C. Identification Tests—General, Chloride 191: A 2-mg/mL solution meets the requirements of the tests.
ASSAY
•  Procedure
Solution A:  2.8 g/L of monobasic sodium phosphate monohydrate and 1.8 g/L of 1-octanesulfonic acid sodium salt monohydrate in water
Mobile phase:  Acetonitrile, methanol, and Solution A (17:24:59)
Diluent:  Use Mobile phase adjusted with diluted hydrochloric acid to a pH of 3.65 ± 0.15.
Standard solution:  1 mg/mL of USP Irinotecan Hydrochloride RS in Diluent
Sample solution:  1 mg/mL of Irinotecan Hydrochloride in Diluent
Chromatographic system 
Mode:  LC
Detector:  UV 255 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Column temperature:  40
Flow rate:  1.5 mL/min
Injection size:  15 µL
System suitability 
Sample:  Standard solution
Suitability requirements 
Tailing factor:  NMT 1.5
Relative standard deviation:  NMT 2.0%
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of irinotecan hydrochloride (C33H38N4O6·HCl) in the portion of Irinotecan Hydrochloride taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak area from the Sample solution
rS== peak area from the Standard solution
CS== concentration of USP Irinotecan Hydrochloride RS in the Standard solution (mg/mL)
CU== concentration of Irinotecan Hydrochloride in the Sample solution (mg/mL)
Acceptance criteria:  98.0%–102.0% on the anhydrous basis
IMPURITIES
•  Residue on Ignition 281: NMT 0.1%
•  Heavy Metals, Method II 231: NMT 10 ppm
•  Limit of Irinotecan Hydrochloride Enantiomer
Mobile phase:  Hexane, dehydrated alcohol, and diethylamine (250:250:1)
Diluent:  Dehydrated alcohol and diethylamine (250:1)
System suitability solution:  0.1 mg/mL each of USP Irinotecan Hydrochloride RS and USP Irinotecan Related Compound D RS in Diluent
Identification solution:  1 mg/mL of USP Irinotecan Hydrochloride RS in Diluent. [Note—This solution is used for Identification test B. ]
Standard solution:  1.5 µg/mL of USP Irinotecan Related Compound D RS in Diluent
Sensitivity solution:  0.5 µg/mL of USP Irinotecan Related Compound D RS in Diluent, from the Standard solution
Sample solution:  1 mg/mL of Irinotecan Hydrochloride in Diluent
Chromatographic system 
Mode:  LC
Detector:  UV 370 nm
Column:  4.6-mm × 25-cm; 10-µm packing L40
Flow rate:  1.0 mL/min
Injection size:  20 µL
System suitability 
Samples:  System suitability solution, Standard solution, and Sensitivity solution
[Note—The relative retention times for irinotecan related compound D (R-enantiomer) and irinotecan (S-enantiomer) are 0.7 and 1.00, respectively. ]
Suitability requirements 
Resolution:  NLT 2.5 between irinotecan related compound D and irinotecan, System suitability solution
Relative standard deviation:  NMT 5.0%, Standard solution
Sensitivity:  The irinotecan related compound D peak should be visible, Sensitivity solution
Analysis 
Samples:   Standard solution and Sample solution
Calculate the percentage of irinotecan hydrochloride R-enantiomer in the portion of Irinotecan Hydrochloride taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak area of irinotecan related compound D from the Sample solution
rS== peak area of irinotecan related compound D from the Standard solution
CS== concentration of USP Irinotecan Related Compound D RS in the Standard solution (mg/mL)
CU== concentration of Irinotecan Hydrochloride in the Sample solution (mg/mL)
Acceptance criteria 
R-enantiomer:  NMT 0.15%
[Note—On the basis of the synthetic route, perform either Organic Impurities Procedure 1 or Organic Impurities Procedure 2. ]
•  Organic Impurities Procedure 1 (for Material Labeled as Produced by a Synthetic Process)
Mobile phase, Diluent, Sample solution, and Chromatographic system:  Proceed as directed in the Assay.
System suitability stock solution:  0.01 mg/mL each of USP Irinotecan Related Compound B RS and USP Irinotecan Related Compound C RS in methanol
System suitability solution:  1.0 µg/mL each of USP Irinotecan Related Compound B RS and USP Irinotecan Related Compound C RS in Diluent, from System suitability stock solution
Standard solution:  2.0 µg/mL of USP Irinotecan Hydrochloride RS in Diluent
Sensitivity solution:  0.5 µg/mL of USP Irinotecan Hydrochloride RS in Diluent
System suitability 
Samples:  System suitability solution, Standard solution, and Sensitivity solution
Suitability requirements 
Resolution:  NLT 1.1 between irinotecan related compound B and irinotecan related compound C, System suitability solution
Relative standard deviation:  NMT 2.0%, Standard solution
Signal-to-noise ratio:  NLT 10, Sensitivity solution
Analysis 
Samples:  Sample solution and Standard solution
Calculate the percentage of each impurity in the portion of Irinotecan Hydrochloride taken:
Result = (rU/rS) × (CS/CU) × 100
rU== peak area of each impurity from the Sample solution
rS== peak area of irinotecan from the Standard solution
CS== concentration of irinotecan hydrochloride in the Standard solution (mg/mL)
CU== concentration of Irinotecan Hydrochloride in the Sample solution (mg/mL)
Acceptance criteria 
Individual impurities:  See Table 1. [Note—Disregard any impurity peaks less than 0.05%. ]
Table 1
Name Relative
Retention
Time
Acceptance
Criteria,
NMT (%)
Irinotecan related compound B 0.55 0.15
Irinotecan related compound C 0.60 0.10
Irinotecan hydrochloride 1.0
Any unspecified impurity 0.10
Total impurities 0.5
•  Organic Impurities Procedure 2 (for Material Labeled as Produced by a Semi-synthetic Process)
Solution A:  2.72 g/L of monobasic potassium phosphate in water. Adjust with dilute phosphoric acid (1 in 20) to a pH of 3.5 ± 0.05.
Solution B:  Acetonitrile and methanol (3:2)
Mobile phase:   See Table 2.
Table 2
Time
(min)
Solution A
(%)
Solution B
(%)
0 80 20
40 30 70
45 30 70
50 80 20
55 80 20
Diluent:  Acetonitrile, methanol, and Solution A (1:1:2)
System suitability solution:  0.1 mg/mL each of USP Irinotecan Hydrochloride RS and USP Irinotecan Related Compound A RS in Diluent
Standard solution:  1 µg/mL of USP Irinotecan Hydrochloride RS in Diluent
Sample solution:  1 mg/mL of Irinotecan Hydrochloride in Diluent
Chromatographic system 
Mode:  LC
Detector:  UV 220 nm
Column:  4.6-mm × 25-cm; 5-µm packing L1
Flow rate:  1 mL/min
Injection size:  10 µL
System suitability 
Samples:  System suitability solution and Standard solution
Suitability requirements 
Resolution:  NLT 3.0 between irinotecan and irinotecan related compound A, System suitability solution
Relative standard deviation:  NMT 2.0%, Standard solution
Analysis 
Samples:  Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Irinotecan Hydrochloride taken:
Result = (rU/rS) × (CS/CU) × (1/F) × 100
rU== peak area of each individual impurity from the Sample solution
rS== peak area of irinotecan from the Standard solution
CS== concentration of USP Irinotecan Hydrochloride RS in the Standard solution (mg/mL)
CU== concentration of Irinotecan Hydrochloride in the Sample solution (mg/mL)
F== relative response factor for each individual impurity (see Table 3)
Acceptance criteria 
Individual impurities:  See Table 3. [Note—Disregard any unspecified impurity peaks less than 0.05%. ]
Table 3
Name Relative
Retention
Time
Relative
Response
Factor
Acceptance
Criteria,
NMT (%)
7-Desethyl irinotecana 0.82 0.77 0.15
Irinotecan 1.00
Irinotecan related compound Ab 1.15 1.4 0.15
11-Ethyl irinotecanc 1.27 0.63 0.15
Camptothecind 1.35 1.4 0.15
Irinotecan related compound Be 1.50 1.3 0.15
7-Ethylcamptothecinf 1.76 1.2 0.15
7,11-Diethyl-10-hydroxycamptothecing 2.05 0.65 0.15
Any unspecified impurity 1.0 0.10
Total impurities 0.50
a  (S)-4-Ethyl-4-hydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione-9-yl (1,4¢-bipiperidine)-1¢-carboxylate.
b  (S)-4-Ethyl-4,9-dihydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
c  (S)-4,8,11-Triethyl-4-hydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione-9-yl (1,4¢-bipiperidine)-1¢-carboxylate.
d  (S)-4-Ethyl-4-hydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
e  (S)-4,11-Diethyl-4,9-dihydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
f  (S)-4,11-Diethyl-4-hydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
g  (S)-4,8,11-Triethyl-4,9-dihydroxy-1H-pyrano[3¢,4¢:6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
SPECIFIC TESTS
•  Microbial Enumeration Tests 61 and Tests for Specified Microorganisms 62: The total aerobic microbial count does not exceed 1000 cfu/g, and the total combined molds and yeasts count does not exceed 100 cfu/g.
•  Water Determination, Method I 921: Between 7.0% and 9.0%
ADDITIONAL REQUIREMENTS
•  Packaging and Storage: Preserve in tight, light-resistant containers, and store at controlled room temperature.
•  Labeling: If a test for Organic Impurities other than Procedure 1 is used, the labeling states the test with which the article complies.
•  USP Reference Standards 11
USP Irinotecan Hydrochloride RS Click to View Structure
USP Irinotecan Related Compound A RS
(S)-4-Ethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
    C20H16N2O5        364.35
USP Irinotecan Related Compound B RS
(S)-4,11-Diethyl-4,9-dihydroxy-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline-3,14(4H,12H)-dione.
    C22H20N2O5        392.40
USP Irinotecan Related Compound C RS
(S)-9-[(1,4'-Bipiperidine)-1'-carbonyloxy]-4-methyl-11-ethyl-3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline hydrochloride.
    C32H36N4O6·HCl        609.11
USP Irinotecan Related Compound D RS
(R)-9-[(1,4'-Bipiperidine)-1'-carbonyloxy]-4,11-diethyl-3,4,12,14-tetrahydro-4-hydroxy-3,14-dioxo-1H-pyrano[3',4':6,7]indolizino[1,2-b]quinoline hydrochloride, trihydrate.
    C33H38N4O6·HCl·3 H2O        677.18
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