Tumor Tracer Isosulphan Blue Process And Quality Study

Isosulphan Blue is the first tumor tracer approved by USFDA for lymphoscintigraphy,which is popularly used for sentinel lymph node location of breast cancer.In this paper,Isosulphan Blue synthesis and quality are studied.The main contents are described as follows:2-chlorobenzaldehyde is used as a key starting material for Isosulphan Blue synthesis,which is firstly sulphonated by 20%oleum to get 2-chloro-l-formylbenzene-5-sulphonic acid sodium salt.Then followed by substitution sulphonation by sodium sulphite and sodium hydrogen sulphite to get 1-formyl benzene-2,5-sulphonic acid disodium salt.Then goes to next step of condensation with N,N-Diethyl aniline to get the 4-[Bis[4-(diethylamino)phenyl]-methyl]benzene-2,5-disulphonic acid disodium salt.Then after oxidation by manganese dioxide to get crude Isosulphan Blue.Then crude product is purified by column purification twice and once recrystallization to get the purified Isosulphan Blue.The process of Isosulphan Blue is optimized to and three process validation batches have been produced.The scale up production results show that Isosulphan Blue process parameters used in production is under control and the product obtained has a good quality to meet its strict product specification.The total yield in production is 11.9%.Based on Isosulphan Blue process and following the current regular guideline,the Isosulphan Blue specification is established,in which,product identification,related substance,metal residue,residue solvents and microbe etc are covered.HPLC Method for Isosulphan Blue related substances testing has been established.Its chromatographic parameters as below:HPLC column is Waters X Terra,RP8 and mobile phase is dipotassium phosphate-Acetonitrile system with gradient elution.The sample solution concentration is 0.5mg/mL and with 10?L injection.The detector is UV220nm and running time is 35 min.Its method validation work has been done and the results show that the method has a good specificity with good resolution between Isosulphan Blue and impurities.The method has good linearity with R>0.99 in the range of 1%-120%.The method accuracy,precision and robustness also meet the method validation requirement.Validation work shows this HPLC method is suitable to Isosulphan Blue related substance testing.HS-GC Method for Isosulphan Blue residue solvents testing has been established too.Its chromatographic parameters as below:GC column ZB-624 is used.Its injector temperature is 180?,detector temperature is 260?,column temperature is from 40?keeping for 7 mins,then increasing to 220? with the rate of 15? per min.Its carrier gas is nitrogen and flow rate is 3mL/min.Headspace Incubation temperature is 80?,Syringe temperature is 90? and incubation time is 20mins.Its injection volume is 1.0mL and running time is 34min.Its method validation work shows that the method has good sensitivity with quantity limit at 173ppm,59ppm,10ppm,6ppm and 0.3ppm for methanol,ethanol,acetone,dichloromethane and n-hexane respectively.The each RSD%of each solvent peak area from 6 injections is less than 5%and each solvent's recovery is between 80-120%and the RSD of 9 recoveries is less than 15%.The method linearity is also checked with good results.The method also has good robustness is also has a good specificity with good resolution between Isosulphan Blue and impurities.The method has good linearity with R>0.99 in the range of 1%-120%.Validation work shows this HS-GC method is suitable to Isosulphan Blue residue solvents testing.Isosulphan Blue Its long term and accelerate stability study shows most of parameters are stable except water content increased a little bit but still meet specification.Isosulphan Blue is stable for 2 years with the package of double PE in aluminium pouch.