title: Quantification of malachite green in fish feed utilising liquid chromatography-tandem mass spectrometry with a monolithic column.
abstract: The purpose of this study was to develop a rapid and sensitive method for the quantification of malachite green (MG) in fish feed using LC-ESI-MS/MS with a monolithic column as stationary phase. Fish feed was cleaned using ultrasonic assisted liquid-liquid extraction. The separation was achieved on a Chromolith® Performance RP-18e column (100 × 4.6 mm) using gradient mobile phase composition of methanol and 0.1% formic acid at the flow rate of 1.0 ml min⁻¹. The analyte was ionised using electrospray ionisation in positive mode. Mass spectral transitions were recorded in selected reaction monitoring (SRM) mode at m/z 329.78 → m/z 314.75 with a collision energy (CE) of 52% for MG. The system suitability responses were calculated for reproducibility tests of the retention time, number of theoretical plates and capacity factor. System validation was evaluated for precision, specificity and linearity of MG. The linearity and calibration graph was plotted in the range of 15.0-250 ng ml⁻¹ with the regression coefficient of >0.997. The lower limits of detection and quantification for MG were 0.55 and 1.44 ng ml⁻¹, respectively, allowing easy determination in fish feed with accuracy evaluated as a percentage recovery of 92.1% and precision determined as % CV of < 5. The method was also extended to the determination of MG in an actual fish feed. The sensitivity and selectivity of LC-ESI-MS/MS using monolithic column offers a valuable alternative to the methodologies currently employed for the quantification of MG in fish feeds.
authors: a Pakistan Council of Scientific and Industrial Research Laboratories Complex , Pharmaceutical Research Center , Karachi Pakistan.
authors: Kamran
authors: K
authors: Abro
authors: Sarfaraz Ahmed
authors: SA
authors: Mahesar
authors: Seema
authors: S
authors: Iqbal
authors: Shahnaz
authors: S
authors: Perveen
mesh_terms: D000821:Animal Feed / Q000032:analysis* / Q000191:economics; D000818:Animals; D000981:Antiprotozoal Agents / Q000032:analysis / Q000737:chemistry; D017756:Aquaculture; D002851:Chromatography, High Pressure Liquid / Q000662:veterinary; D005399:Fishes; D005506:Food Contamination; D005513:Food Inspection / Q000379:methods*; D005659:Fungicides, Industrial / Q000032:analysis / Q000737:chemistry; D057230:Limit of Detection; D059625:Liquid-Liquid Extraction / Q000662:veterinary; D009153:Mutagens / Q000032:analysis* / Q000737:chemistry; D010154:Pakistan; D010573:Pesticide Residues / Q000032:analysis* / Q000737:chemistry; D015203:Reproducibility of Results; D012394:Rosaniline Dyes / Q000032:analysis* / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000662:veterinary; D053719:Tandem Mass Spectrometry / Q000662:veterinary; D013723:Teratogens / Q000032:analysis / Q000737:chemistry; D014465:Ultrasonics
chemical_list: D000981:Antiprotozoal Agents; D005659:Fungicides, Industrial; D009153:Mutagens; D010573:Pesticide Residues; D012394:Rosaniline Dyes; D013723:Teratogens; C005095:malachite green
keywords: LC-ESI-MS/MS; SRM; fish feeds; malachite green; monolithic column