Irinotecan Hydrochloride

This article is compiled based on the United States Pharmacopeia (USP) – 2025 Edition

Issued and maintained by the United States Pharmacopeial Convention (USP)

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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 CAS RN®: 136572-09-3; UNII:  042LAQ1IIS. 

1 DEFINITION 

Irinotecan Hydrochloride contains NLT 98.0% and NMT 102.0% of C₃₃H₃₈N₄O₆.HCl , calculated on the anhydrous basis.

2 IDENTIFICATION 

Change to read: 

Spectroscopic Identification Tests 〈197〉, Infrared Spectroscopy: 197K (CN 1-May-2020)

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. 

Identification Tests—General, Chloride 〈191〉: A 2-mg/mL solution meets the requirements of the tests. 

3 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 

(See Chromatography 〈621〉, System Suitability.) 

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 = (r_U/r_S) × (C_S/C_U) x 100 

r_U = peak area from the Sample solution

r_S = peak area from the Standard solution

C_S = concentration of USP Irinotecan Hydrochloride RS in the Standard solution (µg/mL) 

C_U = concentration of irinotecan hydrochloride in the Sample solution (mg/mL)

Acceptance criteria: 98.0%–102.0% on the anhydrous basis 

4 IMPURITIES 

Residue on Ignition 〈281〉: NMT 0.1% 

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 Identication 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 

(See Chromatography 〈621〉, System Suitability.) 

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 = (r_U/r_S) × (C_S/C_U) x 100 

r_U = peak area of irinotecan related compound D from the Sample solution

r_S = peak area of irinotecan related compound D from the Standard solution

C_S = concentration of USP irinotecan related compound D RS in the Standard solution (µg/mL) 

C_U = 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 = (r_U/r_S) × (C_S/C_U) x 100 

r_U = peak area of each impurity from the Sample solution

r_S = peak area of of irinotecan from the Standard solution

C_S = concentration of USP Irinotecan Hydrochloride RS in the Standard solution (µg/mL) 

C_U = 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 

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 

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 

(See Chromatography 〈621〉, System Suitability.) 

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 = (r_U/r_S) × (C_S/C_U) x (1/F) x 100 

r_U = peak area of each individual impurity from the Sample solution

r_S = peak area of irinotecan from the Standard solution

C_S = concentration of USP Irinotecan Hydrochloride RS in the Standard solution (µg/mL) 

C_U = 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 

^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. 

5 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% 

6 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 

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 

11-Ethyl-4-hydroxy-4-methyl-3,14-dioxo-3,4,12,14-tetrahydro-1H-pyrano[3′,4′:6,7]indolizino[1,2-b]quinolin-9-yl (1,4′-bipiperidine)-1′-carboxylate 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 · 3H₂O 677.18