Fractionation of Acidic, Basic and Neutral Drugs from Urine with an SPE Mixed Mode Strong Anion Exchange Polymeric Resin (Agilent SampliQ-SAX) (Publication #5990-4965EN)
 
Abstract: A polymeric mixed mode strong anion exchange resin, Agilent SampliQ SAX, was evaluated in terms of its ability to extract acidic, basic and neutral drugs from urine. An SPE procedure was applied whereby acidic drugs were eluted in the acidic fraction while the neutral and basic drugs were eluted in the neutral fraction. High recoveries (79.6 – 109%) were obtained. The calibration curves were linear for nortriptyline, ketoprofen and naproxen (r2 > 0.999) in the 0 to 10 μg/mL concentration range. Secobarbital was, however, linear from 0 – 25 μg/mL. The limits of detection were 0.21 μg/mL, 0.04 μg/mL, 0.03 μg/mL, 0.02 μg/mL and quantification values were 0.81 μg/mL, 0.12 μg/mL, 1.04 μg/mL, 2.74 μg/mL for secobarbital, nortriptyline, ketoprofen and naproxen, respectively.
Fractionation of Acidic, Basic and Neutral Drugs from Plasma with an SPE Mixed Mode Strong Cation Exchange Polymeric Resin (Agilent SampliQ-SCX) (Publication #5990-5001EN)
 
Abstract: A method for the simultaneous extraction of drugs (amphetamine, acetaminophen, p-toluamide, m-toluidine and phenobarbital) from spiked human plasma sample was developed. This procedure employs solid phase extraction with a mixed mode strong cation exchange resin, SampliQ SCX. The chromatographic separation and analysis of SPE extracts achieved using 30% methanol and 70% potassium dihydrogen phosphate as a mobile phase under isocratic conditions on an Agilent ZORBAX Eclipse Plus C18 4.6mm x 75mm, 3.5 µm column at 1mL/min flow rate and a diode array detector set at 210 nm. High and reproducible recoveries (> 80%) for all the analytes were obtained. The limits of detection (LOD) and quantification (LOQ) were 0.39 and 0.71 µg/mL for acetaminophen, 0.84 and 1.87 µg/mL for amphetamine, 0.36 and 1.06 µg/mL for m-toluidine, 0.66 and 0.70 µg/mL for p- toluamide, as well as 0.80 and 1.89 µg/mL for phenobarbital, respectively.
 
SampliQ OPT Solid Phase Extraction Sorbent in the Cleanup of Flavonoids in Ginkgo Biloba by HPLC-DAD (Publication #5990-4799EN)
 
Abstract: Sample cleanup of flavonoids (quercetin, kaempferol and isorhamnetin) from Ginkgo biloba tablets was achieved by solid phase extraction employing Agilent's SampliQ Optimized Polymer Technology (OPT) sorbents. Separation of the flavonoids with a 0.5% phosphoric acid/methanol mobile phase was carried out on an Agilent 1200 Series HPLC coupled with a diode array detector (DAD). The HPLC used an Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 75 mm, 3.5 μm) under isocratic conditions and a 9-min total run time. The recoveries for quercetin ranged from 80 to 92% (n=6), kaempferol 83% to 94% (n=6) while those of isorhamnetin ranged from 75% to 102% (n=6), each with %RSD of less than 5. The limits of detection and quantification were 0.57 and 1.91 μg/mL for quercetin, 0.76 and 2.15 μg/mL for kaempferol and 0.88 and 2.55 μg/mL for isorhamnetin, respectively.
 
Determination of Hormones in Fish (Carassuis carassuis) by SampliQ-OPT Solid Phase Extraction with High Performance Liquid Chromatography (Publication # 5990-3845EN)
 
Abstract: Solid-phase extraction (SPE) coupled with high performance liquid chromatography (HPLC) is a necessary component in the workflow of complex food analysis. This note describes the optimization of extraction and quantitative determination of sixteen hormones in meat (estriol, prednisolone, hydrocortisone, prednisone, methylprednisolone, betamethasone, dexamethasone, triamcinolone acetate, gestrinone, prednisolone acetate, hydrocortisone acetate, prednisone acetate, estradiol, cortisone acetate, methyltestosterone, estrone) in crucian carp (Carassius carassius). Results indicate that SPE using an Agilent SampliQ OPT SPE cartridge (60 mg, 3 mL) and HPLC using an Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 250 mm, 5 μm) is suitable for extraction and analysis of these compounds. Recoveries ranged from 76.2 to 106.1 % with relative standard deviations (RSDs) between 1.7 and 8.9 %.
 
Determination of Sulfonamides in Milk Using Solid Phase Extraction and Liquid Chromatography – Tandem Mass Spectrometry (Publication # 5990-3713EN)
 
Abstract: The extraction of trace levels of nine nitrogen-containing sulfa drugs: sulfamethoxazole, sulfadiazine, sulfathiazole, sulfamerazine, sulfamethizole, sulfamethazine, sulfamethoxypyridazine, sulfachloropyridazine, and sulfadimethoxine in milk samples by solid phase extraction was studied using Agilent SampliQ polymeric strong cation exchange cartridges (pSCX) An Agilent 6410 Triple Quadrupole LC/MS-MS was used for the separation and determination of the sulfa drugs. For reversed-phase chromatography, a Zorbax Eclipse Plus column (C18, 3.0 x 50 mm, 1.8 um) with a 0.1% formic acid/acetonitrile gradient was used. Overall recoveries from the milk samples ranged from 73 – 99%, with %RSD values less than 10%. Limits of detection ranged from 0.2-2.0 ng/mL in milk (S/N = 3) depending on the sulfa drug, below the U.S. Food and Drug Administration acceptable levels in milk.
 
Determination of Chloramphenicol, Florfenicol and Thiamphenicol in Honey Using SampliQ OPT Solid-Phase Extraction Cartridges and Liquid Chromatography-Tandem Mass Spectrometry (Publication # 5990-3615EN)
 
Abstract: A method for the simultaneous determination of three antibiotic residues of chloramphenicol (CAP), florfenicol (FF), and thiamphenicol (TAP) in honey has been developed and validated. The analytes are purified by liquid/liquid extraction and solid-phase extraction (SPE) and are quantified by liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) operating in negative ion multiple reaction monitoring (MRM) mode. Chloramphenicol-D5 is used as the internal standard. The method is validated by achieving reproducible, satisfactory, quantitative results. The method provides a sub-ng/g to ng/g level of limit of quantitation (LOQ) for all three antibiotics in honey. The overall recoveries range from 74.9 to 107% with RSD values between 0.5 and 9.7%. The dynamic calibration ranges for chloramphenicol and florfenicol are obtained over 0.1 to 20.0 ng/g and 1.0 to 20.0 ng/g for thiamphenicol. The method is demonstrated to be fast, simple, and efficient for monitoring chloramphenicol, florfenicol, and thiamphenicol residues in honey.
 
Trace-level Analysis of Melamine in Milk Products on Agilent 7890A/5975 GC/MSD Using a New Agilent J&W DB-5ms Ultra Inert Column and SampliQ SCX Cartridges (Publication # 5990-3282EN)
 
Abstract: GC/MS method is presented for the quantitative determination and confirmation of melamine residues in milk products. The milk sample was cleaned up using Agilent’s new SampliQ SCX SPE cartridges before derivatization. The derived extracts were analyzed by GC/MS with EI in synchronous SIM/scan mode on a new Agilent J&W DB- 5ms Ultra Inert column. The method has good linearity in the range of 0.025 to 2.000 μg/mL with a correlation coefficient of 0.9986. The average recoveries of melamine, for milk spiked at 0.080, 0.800, and 1.600 mg/kg, were over 80 percent, with RSDs of less than 5 percent. The LOD is 0.0025 mg/kg in the milk matrix, which is suitable for trace-level analysis of melamine in milk products.
 
Quantitative Liquid Chromatography Analysis of Melamine in Dairy Products Using Agilent’s 1120 Compact LC and 1200 Rapid Resolution LC [and SampliQ SCX SPE Cartridges (Publication # 5989-9949EN)
 
Abstract: In this application, three different LC methods are developed for the determination of melamine in dairy products. The first is a modification of the U.S. FDA method [1]. An Agilent LC system (1120 or 1200) is used with a ZORBAX SB-C8 LC column to run in reversed-phase ion-pair mode for routine quantitation of melamine. The second method is targeted for high throughput using an Agilent Rapid Resolution LC (RRLC) system (1200SL) to speed melamine analysis by more than three times with a Rapid Resolution High Throughput (RRHT) column. The third is an alternative ion-exchange LC method where a ZORBAX 300SCX column is employed to successfully retain melamine using a simple mobile phase of buffered water/acetonitrile without the presence of ion-pair reagent. Due to the complexity of dairy product matrices, a cleanup step using solid phase extraction (SPE) is required for the above methods. The Agilent SampliQ SCX, a mixed-mode polymer SPE cartridge with combined reversed-phase and strong cation exchange properties, is used to successfully remove matrix interferences.
 
Rapid Screening and Confirmation of Melamine Residues in Milk and Its Products by Liquid Chromatography Tandem Mass Spectrometry [and SampliQ Polymer SCX Cartridges (Publication # 5989-9950EN)
 
Abstract: This rapid method uses the Agilent 6410 Triple Quadrupole (QQQ) with a cation ion exchange column for the liquid chromatography tandem mass spectrometry (LC/MS/MS) analysis of dairy products for melamine. Milk and milk products are prepared with a simple SPE cleanup method employing the new Agilent SimpliQ SCX cartridge. The residue is quantified in the multiple reaction monitoring (MRM ) mode. The selectivity of the QQQ can easily eliminate any matrix interferences that may occur in the separation and provide excellent response. The method provides good results with respect to precision, repeatability, and spiked recovery. The recovery of 80 ppb and 50 ppb melamine spikes in milk powder using the external standard calculation is 62.5 and 83.4 percent, respectively, and the RSD is less than 3 percent.
 
Determination of Melamine Residue in Milk Powder and Egg Using Agilent SampliQ Polymer SCX Solid Phase Extraction and the Agilent 1200 Series HPLC/UV (Publication # 5990-3365EN)
 
Abstract: This method was developed for the determination of melamine in milk powder and egg. Solid phase extraction (SPE) and HPLC/UV used is consistent with the Chinese regulatory method. The sample preparation is performed using a polymeric mixed mode strong cation exchange resin. The separation and detection are performed by HPLC/UV. The limit of detection is 10 μg/kg. Linear calibration curves were obtained over a calibration range of 1 to 20 mg/kg. Overall recoveries range from 84 to 96 percent, with RSD values below 3.0 percent.
 
Determination of Penicillins in Meat by High Performance Liquid Chromatography (HPLC/UV) and HPLC/MS/MS (Publication # 5990-3364EN)
 
Penicillins are antibiotics widely used to treat diseases in animals and as a result, are occasionally found in animal products destined for human consumption. In this paper, a solid phase extraction method with a high performance liquid chromatograph tandem mass spectrometer (HPLC/MS/MS) is shown for the simultaneous determination of six antibiotic residues: azlocillin, penicillin G, oxacillin, cloxacillin, nafcillin, and dicloxacillin in animal tissues (porcine muscle). In the method, the reversed phase column Agilent ZORBAX Eclipse Plus C18 (3.5 μm, 100 mm × 2. 1 mm) and an Agilent mixed mode polymer solid phase extraction cartridge (Agilent SampliQ OPT) were combined to give a total solution to the analysis of residual penicillins. The performance of the solid phase extraction procedure on trace residues is quantitatively evaluated by HPLC/MS/MS.
 
Determination of Tetracyclines in Chicken by Solid-Phase Extraction and High Performance Liquid Chromatography (Publication # 5989-9735EN)
 
A method for the simultaneous determination of the 7 antibiotic residues of minocycline, oxytetracycline, tetracycline, demeclocycline, chlortetracycline, methacycline and doxycycline in chicken has been developed. In the method solid phase extraction (SPE) and HPLC/UV consistant with Chinese regulatory methods. Samples are prepared in EDTA-Mcllvaine buffer solution (pH 4.0),the clean up is performed with a SampliQ OPT cartridge, and the HPLC separation is performed with a Zorbax column (5µm,250×4.6mm i.d). The flow rate is 1.5ml/min, the detector wavelength is 350nm, and the injection volume is 100µL. The limits of detection are between 2.5 and 5µg/kg. Linear calibration curves are obtained over the range of 25 and 500µg/kg. Overall recoveries are between 52.0-99.0% with RSD values between 1.0-6.5%.
 
Analysis of Carbaryl and Carbofuran in Drinking Water with Post-Column Derivatization Using Agilent's New LC Column and SampliQ SPE Cartridges (publication # 5989-9802EN)
 
The herbicides carbaryl and carbofuran were analyzed in drinking water samples using Agilent's new reversed-phase columns, Agilent TC-C18(2), and Agilent's new line of SampliQ C18 SPE cartridges. To meet the requirements of EPA Method 531.1 [1], we tried a 400-μL direct injection onto the new column. Even with such a large injection volume, the new column provided good resolution from interfering or coeluting compounds, as well as highly symmetrical peaks and a high performance result, which gives high sensitivity. In addition, post-column derivatization and a fluorescence detector were used, making the limit of detection (LOD) less than 0.01 ng on column, a lower LOD than that specified in China's drinking water standards [2]. This method used new Agilent SampliQ C18 cartridges to concentrate the water samples and made it possible to determine carbaryl and carbofuran in drinking water at the ppt level. This total solution is very suitable for trace-level analysis of carbaryl and carbofuran in drinking water.
 
Determination of Fungicides in Apple Juice by SampliQ-SCX Solid Phase Extraction with High Performance Liquid Chromatography (Publication # 5990-3235EN)
 
Solid-phase extraction (SPE) coupled with high-performance liquid chromatography (HPLC) was optimized for extraction and quantification of two fungicides (carbendazim and thiabendazole) in apple juice. Results indicate that SPE using Agilent SampliQ SCX (60 mg, 3 mL) and HPLC using an Agilent ZORBAX Eclipse Plus C18 column (4.6 mm × 100 mm, 3.5 μm) is an excellent combination for extraction and analysis of these compounds. Recoveries ranged from 92.1 to 99.4 percent with RSDs below 5 percent and limits of detection of 4 μg/kg.
 
Determination of Multi Residue Tetracycline and their metabolites in Milk by High Performance Liquid Chromatography – Tandem Mass Spectrometry (Publication # 5990-3816EN)
 
Tetracyclines are probably the most frequently used antibiotics in animal husbandry. In this paper, a high performance liquid chromatography tandem mass spectrometric (HPLC /MS/MS) method is developed for the simultaneous determination of 10 antibiotic residues: minocycline, 4-epioxytetracycline, 4-epitetracycline, tetracycline, 4-epichlortetracycline, demeclocycline, chlortetracycline, methacycline, doxycycline, oxytetracycline in milk and animal tissues. In the method, Agilent’s novel solid phase extraction cartridge and a reversed phase Agilent ZORBAX RX C8 column (5 μm, 150 mm × 2.1 mm) are used for purification and separation. The limit of detection (LOD) is between 0.5 and 10.0 μg/kg and the limit of quantitation (LOQ) is less than 50 μg/kg. The linearity is obtained from 5 to 1000 μg/kg. Overall recoveries are between 76.4% and 101% with a relative standard deviation (RSD, n = 6) less than 8.4%. The method is rapid, sensitive, convenient and robust, and can be used to simultaneously confirm multi-residues of tetracyclines and their metabolites in milk.
 
Determination of ОІ2-Agonists in Pork Using Agilent SampliQ SCX Solid-Phase Extraction Cartridges and Liquid Chromatography-Tandem Mass Spectrometry (Publication # 5990-4180EN)
 
A method for simultaneous determination of four β2-agonist residues of terbutaline, salbutamol, clenbuterol and formoterol in pork has been developed and validated. The analytes are purified by liquid-liquid extraction (LLE) and solid-phase extraction (SPE) and quantified by liquid chromatography coupled to electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) operating in positive ion multiple reaction monitoring (MRM) mode. The method provides a sub-ng/g level of limit of detection (LOD) for all four β2-agonists in pork. The dynamic calibration ranges for these compounds are obtained from 0.25 to 5 ng/g. The overall recoveries range from 78 to 101% with RSD values between 1.8 and 7.2%.
 
High Speed, Ultra-High Sensitivity, and Robustness Needed for the Quantitation of Pharmaceuticals in Blood Plasma (Publication #5990-3823EN)
 
Abstract: This Application Note demonstrates the performance of an Agilent 1200 Series Rapid Resolution LC (RRLC) system coupled to an Agilent 6460A Triple Quadrupole (QQQ) mass spectrometer for the bioanalysis of human plasma. High throughput analysis is shown for a test pharmaceutical and an internal standard with both drugs eluting well within a 1 minute chromatographic window. Ultra-high sensitivity is demonstrated by linearity across a concentration range typically covered in human microdosing pharmacokinetic studies (1–500 pg/mL plasma). Robustness and reproducibility are shown for this concentration range over a 5-day period during which the LC/MS system was exposed to more than 1000 plasma samples.
 
Agilent SampliQ OPT Solid-Phase Extraction Sorbent in the Clean-up of Alkaloids in Goldenseal by HPLC-DAD (Publication #5990-4188EN)
 
Abstract: Sample clean-up of alkaloids of Goldenseal commercial products (hydrastine and berberine) was achieved by solid phase extraction employing Agilent's SampliQ Optimized Polymer Technology (OPT) sorbents. Separation of the products with a 0.1% phosphoric acid/methanol mobile phase was carried out on an Agilent 1200 series HPLC coupled with a diode array detector (DAD) on an Agilent ZORBAX Eclipse Plus C18 column (4.5 mm × 75 mm × 3.5 μm) using gradient elution with a 6 min total run time. The recovery for hydrastine ranged from 101% to 106% (n = 8) while that of berberine ranged from 71% to 82%, (n = 8), each with % relative standard deviation (RSD) of less than 1. The limits of detection and quantification for hydrastine were 0.50 and 1.65 μg/mL respectively while those of berberine were 0.47 and 1.55 μg/mL, respectively. Goldenseal sample from Willow contained 17 μg/mL hydrastine and 35 μg/mL berberine while Goldenseal sample from Solga contained 6 μg/mL hydrastine and 12 μg/mL berberine.
 
 
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