A liquid chromatographic method for the determination of histamine in immunoglobulin preparation using solid phase extraction and pre-column derivatization.
An immunoglobulin (IgG) preparation with micro-amount of histamine fixed on the active protein fraction has been used to increase the resistance to allergic reactions. However, excessive histamine may cause hypo- or hypertension, headache, or anaphylactic shock and so the histamine content of the drug is strictly controlled by a regulation: 0.15 microg of histamine dihydrochloride is allowed for 12 mg of immunoglobulin. In this study, a liquid chromatographic method to determine micro-amount of histamine in the pharmaceutical was developed and validated. This method include a sample cleanup by a solid phase extraction (SPE) using a polystyrene-divinyl benzene (PS-DVB) polymeric sorbent and high-performance liquid chromatography after precolumn fluorescent labeling of the histamine with o-phthaldialdehyde. The drug sample was loaded to the SPE cartridge after adjusting to pH 9.5. After successive washings of the cartridge with water and 30% aqueous methanol, histamine was then eluted with 100 mM sodium acetate (pH 9.5)-methanol (20:80, v/v). An aliquot from the eluate was labeled with o-phthaldialdehyde-mercaptoethanol (OPA-ME) for fluorescence detection at the excitation maximum of 340 nm and emission maximum of 450 nm. HPLC analysis was performed on a phenyl-hexyl column with an acetonitrile-phosphate buffer (pH 6.8; 50 microM) (35:65, v/v) as the mobile phase. The retention times of histamine and 3-methylhistamine (IS) were approximately 7.2 and 9.5 min, respectively. The quantitation range was between 0.01-0.2 mg/mL of histamine showing good linearity (r=0.9996). This analytical method would provide a potential mean for the strict control of histamine content in the pharmaceutical product.
Kim
Nam Hee
NH
College of Pharmacy CBITRC, Chungbuk National University, Cheongfu 361-763, Korea.
Park
Youmie
Y
Jeong
Eun Sook
ES
Kim
Chang-Soo
CS
Jeoung
Min Kyo
MK
Kim
Kyoung Soon
KS
Hong
Seung-Hwa
SH
Son
Jong-Keun
JK
Hong
Jin Tae
JT
Park
Il-Young
IY
Moon
Dong-Cheul
DC
D000097:Acetonitriles / Q000737:chemistry; D018926:Anti-Allergic Agents / Q000737:chemistry* / Q000592:standards; D002021:Buffers; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000592:standards; D006632:Histamine / Q000032:analysis*; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry*; D007202:Indicators and Reagents / Q000737:chemistry; D007475:Ion Exchange Resins / Q000737:chemistry; D008623:Mercaptoethanol / Q000737:chemistry; D000432:Methanol / Q000737:chemistry; D008761:Methylhistamines / Q000032:analysis; D011137:Polystyrenes / Q000737:chemistry; D011786:Quality Control; D015203:Reproducibility of Results; D019346:Sodium Acetate / Q000737:chemistry; D052616:Solid Phase Extraction; D012997:Solvents / Q000737:chemistry; D013050:Spectrometry, Fluorescence; D009764:o-Phthalaldehyde / Q000737:chemistry
D000097:Acetonitriles; D018926:Anti-Allergic Agents; D002021:Buffers; D007074:Immunoglobulin G; D007202:Indicators and Reagents; D007475:Ion Exchange Resins; D008761:Methylhistamines; D011137:Polystyrenes; D012997:Solvents; D019346:Sodium Acetate; D008623:Mercaptoethanol; D009764:o-Phthalaldehyde; C031205:3-methylhistamine; D006632:Histamine; C003771:Amberlite XAD-2 resin; D000432:Methanol; C032159:acetonitrile