Published on USP Medicines Compendium (https://www.usp-mc.

org)

Dihydroartemisinin and Piperaquine Phosphate Tablets

For Development Version 0.1

DEFINITION
Dihydroartemisinin and Piperaquine Phosphate Tablets contain NLT 95.0% and NMT 105.0% of the labeled amount of
dihydroartemisinin (C15H24O5) and piperaquine phosphate [C29H32Cl2N6 · 4(H3PO4)].

Performance-Based Monograph
(Contains tests, procedures, and acceptance criteria for the material under test. It also includes the criteria-based procedures that
define the equivalence criteria necessary to demonstrate that an Acceptable Procedure is equivalent to the Reference Procedures.)

IDENTIFICATION
• A. The response for piperaquine and dihydroartemisinin (α and β) from the Sample solution corresponds to that of the Standard
solution, as obtained in the Assay.

ASSAY
• PROCEDURE
Standard solution: MC α-Dihydroartemisinin CRM and MC Piperaquine Phosphate CRM in an appropriate diluent
Sample solution: Dilute a portion of Tablets in an appropriate diluent to obtain a concentration approximately the same as that of
the Standard solution.
Analytical system: Use a procedure validated as described in MC general chapter Assessing Validation Parameters for Reference
and Acceptable Procedures <10>.
System performance requirements
Precision: Meet the requirements for 98.0%– 102.0% for each component
Accuracy: Meet the requirements for 98.0%– 102.0% for each component
Specificity: Meet the requirements
Range: Meet the requirements
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of the labeled amount of dihydroartemisinin (C15H24O5) and piperaquine phosphate [C29H32Cl2N6 · 4(H3PO4
)] in the Sample solution:

Result = (rU/rS) × (CS/CU) × 100

rU = peak response from the Sample solution [NOTE—Add responses from the α and β forms of dihydroartemisinin.]
rS = peak response from the Standard solution
CS = concentration of MC α-Dihydroartemisinin CRM and MC Piperaquine Phosphate CRM in the Standard solution
CU = nominal concentration of dihydroartemisinin and piperaquine phosphate in the Sample solution
Acceptance criteria: 95.0%–105.0% of the labeled amount of each component

PERFORMANCE TESTS
• DISSOLUTION <711>
Medium, Apparatus, Time, and Acceptance criteria: See MC general chapter Assessing Dosage Form Performance Quality
<12>.
Standard solution: MC α-Dihydroartemisinin CRM and MC Piperaquine Phosphate CRM in a ratio matching the label claim of the
Tablets in an appropriate diluent
Sample solution: Pass a portion of the solution under test through a filter of 0.45-µm pore size, and dilute with an appropriate
diluent to obtain a concentration approximately the same as that of the Standard solution.
Instrumental conditions: Proceed as directed in the Assay or in an alternatively validated procedure.
• UNIFORMITY OF DOSAGE UNITS <905>: Meet the requirements

IMPURITIES
• ELEMENTAL IMPURITIES <232>: Proceed as directed in the chapter.
 • RESIDUAL SOLVENTS <467>: Proceed as directed in the chapter.
• ORGANIC IMPURITIES
Standard solution: In a ratio matching the label claim of the Tablets and all appropriate MC Impurity RMs, at concentrations
corresponding to the Acceptance criteria of the impurity, in an appropriate diluent
Sample solution: Dilute a portion of Tablets in an appropriate diluent to obtain a concentration approximately the same as that of
the Standard solution.
Analytical system: Use a procedure validated as described in MC general chapter Assessing Validation Parameters for Reference
and Acceptable Procedures <10>.
System performance requirements
Precision: Meet the requirements
Accuracy: Meet the requirements
Ruggedness: Meet the requirements
Specificity: Meet the requirements
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each impurity in the Sample solution:

Result = (rU/rS) × (CS/CU) × 100

rU = peak response of each impurity from the Sample solution

rS = peak response of each MC Impurity RM from the Standard solution. [NOTE—If no MC Impurity RMs are available,
use the peak response of dihydroartemisinin or piperaquine, whichever gives the most precise results.]
CS = concentration of standard material in the Standard solution
CU = nominal concentration of dihydroartemisinin and piperaquine phosphate in the Sample solution
Acceptance criteria
Any individual impurity: NMT 0.2%
Total Impurities: NMT 1.0%

SPECIFIC TESTS
• WATER DETERMINATION, Method Ia <921>
Sample: Powder NLT 5 Tablets, and use about 200 mg, accurately weighed.
Acceptance criteria: NMT 7.5%

ADDITIONAL REQUIREMENTS
• REFERENCE STANDARDS <11>
MC α-Dihydroartemisinin CRM
MC Dihydroartemisinin Impurity A RM
(2R)-2-[(4R)-4-Methyl-2-oxo-3-(3-oxobutyl)cyclohexyl]propanal.
MC Dihydroartemisinin Impurity B RM
(2R)-2-[(4R)-4-Methyl-2-oxo-3-(3-oxobutyl)cyclohexyl]propionic acid.
MC Dihydroartemisinin Impurity C RM
(3R,5aS,6R,8aS,9R,12S,12aR)-Octahydro-3,6,9-trimethyl- 3,12-epoxy-12H-pyrano[4,3-j]-1,2-benzodioxepin-10(3H)-one.
MC Dihydroartemisinin Impurity D RM
(3R,3aS,6R,6aS,9R,10aS)-3,6,9-Trimethyloctahydro-9,10b-epoxyoxepino[4,3,2-ij]isochromen-2(3H,10aH)-one.
MC β-Dihydroartemisinin RM
MC Piperaquine Phosphate CRM
MC Piperaquine Impurity A RM
4-Piperazin-7chloro-quinoline.
MC Piperaquine Impurity B RM
7-Chloro-4-{4-[3-(piperazin-1-yl)propyl]piperazin-1-yl}quinoline.
MC Piperaquine Impurity C RM
5-Chloro-4-(4-{3-[4-(7-chloroquinolin-4-yl)piperazin-1-yl]propyl}piperazin-1-yl)quinoline.
MC Piperaquine Impurity D RM
7-Chloroquinolin-4-ol.
MC Piperaquine Impurity E RM
4,4’-[Piperazine-1,4-diylbis(propane-3,1-diylpiperazine-4,1-diyl)]bis(7-chloroquinoline).
 MC Piperaquine Impurity F RM
7-Chloro-4-(4-{3[(7-chloroquinolin-4-yl)oxy]propyl}piperazin-1-yl)quinoline.
MC Piperaquine Impurity G RM
N-(7-Chloroquinolin-4-yl)-N’-{3-[4-(7-chloroquinolin-4-yl)piperazin-1-yl]propyl}ethane-1,2-diamine.
MC Piperaquine Impurity H RM
4,4’-Piperazine-1,4diylbis(7-chloroquinoline).
MC Piperaquine Impurity I RM
7-Chloro-4-[4-(3-chloropropyl)piperazin-1-yl]quinoline.

REFERENCE PROCEDURES
(This section provides detailed descriptions of procedures that may be used for the evaluation of the material under test. These
procedures have been fully validated and the data is available on the MC website.)

TO COME

ACCEPTABLE PROCEDURES
(This section provides detailed descriptions of procedures that may be used for the evaluation of the material under test. These
procedures meet the requirements of the Performance-Based Monograph portion of this standard. Validation data have been provided
that demonstrate the acceptability, and therefore equivalence, of these procedures and the Reference Procedures. Before using this
procedure, a user must demonstrate that the procedure will provide acceptable results with their drug substance by meeting the
requirements of the criteria-based procedures presented in the Performance-Based Monograph.)

ASSAY
• CONTENT OF PIPERAQUINE PHOSPHATE
Solution A: 13 mM trifluoroacetic acid in water.
Solution B: 13 mM trifluoroacetic acid in methanol and acetonitrile (1:1)
Diluent: 13 mM trifluoroacetic acid in water
System suitability solution: 1.0 mL/mL of MC Piperquine Phosphate CRM an 0.1 mg/mL of each MC Piperaquine Impurity RM in
Diluent
Standard solution: 1.0 mg/mL of MC Piperaquine Phosphate CRM in Diluent
Sample solution: Powdered Tablets equivalent to 1.0 mg/mL of Piperaquine Phosphate in Diluent
Chromatographic system
(See Chromatography <621>, System Suitability.)
Mode: LC
Detector: UV 240 nm
Column: 4.6-mm × 150-cm; 3µm packing L1 (similar to Ace C-18)
Column temperature: 40°
Flow rate: 1.5 mL/min
Injection volume: 5 µL
Mobile phase: See Table 1.

Table 1

Time
Solution A (%) Solution B (%)
(min)

0 85 15

8 82 18

14 82 18

17 70 30

21 70 30

21.1 85 15

30 85 15

System suitability
 Sample: System suitability solution
Suitability requirements
Resolution: NLT 1.5 between piperaquine impurity D and piperaquine
Relative standard deviation: NMT 1.0% for piperaquine phosphate
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of piperaquine phosphate [C29H32Cl2N6 · 4(H3PO4)] in the portion of Piperaquine Phosphate taken:

Result = (rU/rS) × (CS/CU) × 100

rU = peak response of piperaquine phosphate from the Sample solution

rS = peak response of piperaquine phosphate from the Standard solution
CS = concentration of MC Piperaquine Phosphate CRM in the Standard solution. [NOTE—The potency of the Reference
Material and water content is included in this term.]
CU = nominal concentration of Piperaquine Phosphate in the Sample solution [NOTE—Correct for water content in this
term.]

• CONTENT OF DIHYDROARTEMISININ
Buffer: 0.01 M of ammonium phosphate in water
Mobile phase: Buffer, methanol, and acetonitrile (5:3:2). Adjust with 1 M phosphoric acid to a pH of 4.
System suitability solution A: 4 mg/mL of MC α-Dihydroartemisinin CRM, 0.12 mg/mL of MC Dihydroartemisinin Impurity A RM,
0.08 mg/mL of MC Dihydroartemisinin Impurity B RM, and 0.04 mg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
System suitability solution B: 4 µg/mL of MC α-Dihydroartemisinin CRM in acetonitrile
Standard solution: 4 mg/mL of MC α-Dihydroartemisinin CRM in acetonitrile
Sample solution: Powdered Tablets equivalent to about 4 mg/mL of Dihydroartemisinin in acetonitrile
Chromatographic system
(See Chromatography <621>, System Suitability.)
Mode: UHPLC
Detector: UV 210 nm
Column: 2.1-mm × 50-mm; 2.2 µm packing L1 (similar to Acclaim C18)
Column temperature: 10°
Flow rate: 0.5 mL/min
Injection volume: 2 µL
System suitability
Sample: System suitability solution A and System suitability solution B
Suitability requirements
Resolution: NLT 2.5 between impurity A and α-dihydroartemisinin, System suitability solution A
Relative standard deviation: NMT 1.0% for the sum of the α and β forms of dihydroartemisinin, System suitability solution A
Signal to noise ratio: NLT 10 for replicate injections of System suitability solution B
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of dihydroartemisinin (C15H24O5) in the portion of Dihydroartemisinin taken:

Result = (rU/rS) × (CS/CU) × 100

rU = sum of the peak responses of α and β forms of dihydroartemisinin from the Sample solution
rS = peak response of α-dihydroartemisinin from the Standard solution
CS = concentration of α-dihydroartemisinin in the Standard solution. [NOTE—The potency of the Reference Material is included in
this term.]
CU = nominal concentration of dihydroartemisinin in the Sample solution

PERFORMANCE TESTS
• DISSOLUTION OF PIPERAQUINE PHOSPHATE
(See Dissolution <711>.)
Medium: Simulated gastric fluid without pepsin; 900 mL
Apparatus 2: 50 rpm
 Time: 15 min
Acceptance criteria: NLT 80% (Q) of labeled amount
Standard solution: 0.1 mg/mL of MC Piperaquine Phosphate CRM in medium
Sample solution: Pass a portion of the solution under test through a filter of 0.45-µm pore size, and dilute with an appropriate
diluent to obtain a concentration approximately the same as that of the Standard solution.
Instrumental conditions: Proceed as directed in the Acceptable Procedures, Assay
• DISSOLUTION OF DIHYDROARTEMISININ
(See Dissolution <711>.)
Medium: 0.5% sodium dodecylsulfate in 0.01 M monobasic sodium phosphate (pH 6.8); 900 mL
Apparatus 2: 100 rpm
Time: 60 min
Acceptance criteria: NLT 70% (Q) of the labeled content
Mobile phase: 0.1% formic acid in acetonitrile and water (4:1)
System suitability solution: 10.5 µg/mL of MC α-Dihydroartemisinin CRM in Medium
Standard solution 1: 5.25 µg/mL of MC α-Dihydroartemisinin CRM in Medium
Standard solution 2: 8.4 µg/mL of MC α-Dihydroartemisinin CRM in Medium
Standard solution 3: 10.5 µg/mL of MC α-Dihydroartemisinin CRM in Medium
Standard solution 4: 12.6 µg/mL of MC α-Dihydroartemisinin CRM in Medium
Sample solution: Pass a portion of the solution under test through a filter of 0.45-µm pore size, and dilute 0.300 mL of the sample
with 0.900 mL of Medium.
Chromatographic system
(See Chromatography <621>, System Suitability.)
Mode: LC
Detector: MS/MS (Selected Reaction Monitoring)
Molecular Ion: 267 amu
Daughter Ion: 163 amu
Column: 4.6-mm × 150-cm; 3.5 µm packing L1 (similar to Sunfire C18)
Column temperature: 10°
Flow rate: 1 mL/min
Injection volume: 15 µL
System suitability
Sample: System suitability solution
Suitability requirements
Relative standard deviation: NMT 1.0% for the sum of the α and β forms of dihydroartemisinin, System suitability solution
Analysis
Samples: Standard solution 1, Standard solution 2, Standard solution 3, Standard solution 4, and Sample solution
Create a calibration curve using the daughter ion responses from Standard solutions 1–4. Determine the content of
Dihydroartemisinin using the calibration curve.
• UNIFORMITY OF DOSAGE UNITS <905>: Meet the requirements for each drug substance

IMPURITIES
• ORGANIC IMPURITIES, PIPERAQUINE PHOSPHATE IMPURITIES
Solution A, Solution B, Diluent, Mobile phase, System suitability solution, Standard solution, Sample solution,
Chromatographic system, and System suitability: Proceed as directed in Acceptable Procedures, Assay, Content of Piperaquine
Phosphate.
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Piperaquine Phosphate taken:

Result = (rU/rS) × (CS/CU) × F × 100

rU = peak response of each impurity from the Sample solution

rS = peak response of each impurity from the Standard solution. [NOTE—Where an impurity other than those included in the
Standard solution is found in the Sample solution, the peak response from the nearest identified impurity is used for the
calculation.]
CS = concentration of each impurity in the Standard solution. [NOTE—the potency of the Reference Material is included in this
 term.]
CU = nominal concentration of Piperaquine Phosphate in the Sample solution
F = relative response factors (see Table 2)

Table 2

Approx.
Relative
Retention
Name Response
Time
Factor
(min)

Piperaquine impurity A 2.7 0.53

Piperaquine impurity B 3.2 0.83

Piperaquine impurity C 9.9 0.72

Piperaquine impurity D 10.9 0.40

Piperaquine impurity E 14.1 0.75

Piperaquine impurity F 17.3 0.55

Piperaquine impurity G 17.8 0.65

Piperaquine impurity H 19.9 0.70

• ORGANIC IMPURITIES, LIMIT OF DIHYDROARTEMISININ IMPURITY A

Buffer, Mobile phase, System suitability solution A, System suitability solution B, Standard solution, Sample solution,
Chromatographic system, and System suitability: Proceed as directed in Acceptable Procedures, Assay, Content of
Dihydroartemisinin.
Analysis
Samples: Standard solution and Sample solution
Calculate the percentage of each impurity in the portion of Dihydroartemisinin taken:

Result = (rU/rS) × (CS/CU) × F x 100

rU = peak response of each impurity from the Sample solution

rS = peak response of each impurity from the Standard solution. [NOTE—Where an impurity other than those included in the
Standard solution is found in the Sample solution, the peak response from the nearest identified impurity is used for the
calculation.]
CS = concentration of each impurity in the Standard solution. [NOTE—the potency of the Reference Material is included in this
term.]
CU = nominal concentration of Dihydroartemisinin in the Sample solution
F = relative response factor (see Table 3)

Table 3

Relative Relative
Name Retention Response
Time Factor

Dihydroartemisinin
impurity A 0.25 0.196

Dihydroartemisinin
impurity B 0.35 —

Dihydroartemisinin
impurity C 0.75 —

Dihydroartemisinin
impurity D 0.82 —

• ORGANIC IMPURITIES, LIMIT OF DIHYDROARTEMISININ IMPURITIES B, C, AND D

 Mobile phase: 13 mM trifluoroacetic acid in water, methanol, and acetonitrile (5:3:2)
Standard solution B1: 0.2 µg/mL of MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B2: 0.8 µg/mL of MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B3: 1.6 µg/mL of MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B4: 2.4 µg/mL of MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B5: 3.2 µg/mL of the MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B6: 4.0 µg/mL of the MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution B7: 6.0 µg/mL of MC Dihydroartemisinin Impurity B RM in acetonitrile
Standard solution C1: 0.2 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C2: 0.4 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C3: 0.6 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C4: 0.8 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C5: 1.0 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C6: 1.2 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution C7: 1.4 µg/mL of MC Dihydroartemisinin Impurity C RM in acetonitrile
Standard solution D1: 0.2 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D2: 2.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D3: 4.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D4: 8.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D5: 10.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D6: 12.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Standard solution D7: 14.0 µg/mL of MC Dihydroartemisinin Impurity D RM in acetonitrile
Sample solution: Powdered Tablets equivalent to about 4 mg/mL of Dihydroartemisinin in acetonitrile
Chromatographic system
(See Chromatography <621>, System Suitability.)
Mode: UHPLC
Detector: MS/MS (Selected Reaction Monitoring)
Impurity B
Molecular ion: 253 amu
Daughter ion: 183 amu
Impurity C
Molecular ion: 267 amu
Daughter ion: 203 amu
Impurity D
Molecular ion: 211 amu
Daughter ion: 175 amu
Column: 2.1-mm × 50-mm; 2.2 µm packing L1 (similar to Acclaim C18)
Column temperature: 10°
Flow rate: 0.5 mL/min
Injection volume: 2 µL
System suitability
Sample: Standard solution B5, Standard solution C4, and Standard solution D5
Suitability requirements
Relative standard deviation
NMT 10.0% for Impurity B, Standard solution B5
NMT 15.0% for Impurity C, Standard solution C4
NMT 5.0% for Impurity D, Standard solution D5
Analysis
Samples: Standard solutions B1–B7, Standard solutions C1–C7, Standard solutions D1–D7, and Sample solution
Create a calibration curve using the daughter ion responses for dihydroartemisinin impurity B, dihydroartemisinin impurity C, and
dihydroartemisinin impurity D from Standard solutions B1–B7, Standard solutions C1–C7, Standard solutions D1–D7, respectively.
Determine the content of each dihydroartemisinin impurity using the calibration curves.

Source URL (modified on 2012/11/15 - 7:09pm): https://www.usp-mc.org/monographs/dihydroartemisinin-and-piperaquine-phosphate-tablets-0-1