Enhancement of atmospheric pressure chemical ionization for the determination of free and glycine-conjugated bile acids in human serum

A highly sensitive and accurate method based on the precolumn derivatization of bile acids (BA) with a high ionization efficiency labeling reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl-benzenesulfonate (BDEBS) coupled with LC/MS has been developed. After derivatization, BA molecules introduced a weak basic nitrogen atom into the molecular core structure that was readily ionized in commonly used acidic HPLC mobile phases. Derivatives were sufficiently stable to be efficiently analyzed by atmospheric pressure chemical ionization (APCI)-MS/MS in positive-ion mode. The MS/MS spectra of BA derivatives showed an intense protonated molecular ion at m/z [M + H](+). The collision-induced dissociation of the molecular ion produced fragment ions at [MH - H2O](+), [MH - 2H(2)O](+), [MH - 3H(2)O](+). The characteristic fragment ions were at m/z 320.8, 262.8, and 243.7 corresponding to a cleavage of N - CO, O - CO, and C - OCC, respectively, and bonds of derivatized molecules. The selected reaction monitoring, based on the m/z [M + H]+ -> [MH - H2O](+), [MH - H2O](+), [MH - 2H(2)O](+), [MH-3H(2)O](+), 320.8, 262.8, and 243.7 transitions, was highly specific for the BA derivatives. The LODs for APCI in a positive-ion mode, at an S/N of 5, were 44.36-153.6 fmol. The validation results showed high accuracy in the range of 93-107% and the mean interday precision for all standards was < 15% at broad linear dynamic ranges (0.0244-25nmol/mL). Good linear responses were observed with coefficients of > 0.9935 in APCI/MS detection. Therefore, the facile BDEBS derivatization coupled with mass spectrometric analysis allowed the development of a highly sensitive and specific method for the quantitation of trace levels of the free and glycine-conjugated BA from human serum samples.

Development of a sensitive fluorescent derivatization reagent 1,2-benzo-3,4-dihydrocarbazole-9-ethyl chloroformate (BCEOC) and its application for determination of amino acids from seeds and bryophyte plants using high-performance liquid chromatography with fluorescence detection and identification with electrospray ionization mass spectrometry

A pre-column derivatization method for the sensitive determination of amino acids and peptides using the tagging reagent 1,2-benzo-3,4dihydrocarbazole-9-ethyl chloroformate (BCEOC) followed by high-performance liquid chromatography with fluorescence detection has been developed. Identification of derivatives was carried out by liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS/MS). The chromophore of 2-(9-carbazole)-ethyl chloroformate (CEOC) reagent was replaced by 1,2-benzo-3,4-dihydrocarbazole functional group, which resulted in a sensitive fluorescence derivatizing reagent BCEOC. BCEOC can easily and quickly label peptides and amino acids. Derivatives are stable enough to be efficiently analyzed by high-performance liquid chromatography. The derivatives showed an intense protonated molecular ion corresponding m/z (M + H)(+) under electrospray ionization (ESI) positive-ion mode with an exception being Tyr detected at negative mode. The collision-induced dissociation of protonated molecular ion formed a product at m/z 246.2 corresponding to the cleavage of C-O bond of BCEOC molecule. Studies on derivatization demonstrate excellent derivative yields over the pH 9.0-10.0. Maximal yields close to 100% are observed with a 3-4-fold molar reagent excess. Derivatives exhibit strong fluorescence and extracted detzvatization solution with n-hexane/ethyl acetate (10:1, v/v) allows for the direct injection with no significant interference from the major fluorescent reagent degradation by-products, such as 1,2-benzo-3,4-dihydrocarbazole-9-ethanol (BDC-OH) (a major by-product), mono- 1,2-benzo-3,4-dihydrocarbazole-9-ethyl carbonate (BCEOC-OH) and bis-(1,2-benzo-3,4-dihydrocarbazole-9-ethyl) carbonate (BCEOC)(2). In addition, the detection responses for BCEOC derivatives are compared to those obtained with previously synthesized 2-(9-carbazole)-ethyl chloroformate (CEOC) in our laboratory. The ratios AC(BCEOC)/AC(CEOC) = 2.05-6.51 for fluorescence responses are observed (here, AC is relative fluorescence response). Separation of the derivatized peptides and amino acids had been optimized on Hypersil BDS C-18 column. Detection limits were calculated from 1.0 pmol injection at a signal-to-noise ratio of 3, and were 6.3 (Lys)-177.6 (His) fmol. The mean interday accuracy ranged from 92 to 106% for fluorescence detection with mean %CV < 7.5. The mean interday precision for all standards was < 10% of the expected concentration. Excellent linear responses were observed with coefficients of > 0.9999. Good compositional data could be obtained from the analysis of derivatized protein hydrolysates containing as little as 50.5 ng of sample. Therefore, the facile BCEOC derivatization coupled with mass spectrometry allowed the development of a highly sensitive and specific method for the quantitative analysis of trace levels of amino acids and peptides from biological and natural environmental samples. (c) 2005 Elsevier B.V. All rights reserved.

HMLH1 gene mutation in gastric cancer patients and their kindred

AIM: To study the status of hMLH1 gene point mutations of gastric cancer kindreds and gastric cancer patients from northern China, and to find out gene mutation status in the population susceptible to gastric cancer.

Memorias políticas y militares, para servir de continuacion a los Comentarios del Marques de San Felipe : desde el año de MDCCXXV, en que concluyó este autor su obra, hasta el presente, con los Tratados de Paz Y Alianzas de España correspondientes

Signaturas: [ ]1, A-Z4, 2A-2R4, 2S1.

Coronica general de la Orden de San Benito, Patriarca de Religiosos

Colofón en v.1, v.2, v.5, v.6, y v.7.

Instruccion y Reglas de la Congregacion de San Luis Gonzaga

Signaturas: A-Q8/4.

Cuaderno de diseño de las asesorias de talla en piedra en San Agustin y José Isnos Huila, Barichara Santander, forja en hierro en Bogotá y moldes en resina poliester reforzada con fibra de vidrio Barranquilla y Galapa Atlántico.

120 hojas : ilustraciones, fotografías.

Dulcería tradicional en San Jacinto y Montería, forja y cantería en Cartagena, talla madera en Galapa, jabón de tierra en Coyaima, trabajo en guadua en Palo Cabildo. Cuaderno de diseño.

160 hojas : ilustraciones, fotocopias.

Cuaderno de diseño de las asesorías de cestería en paja tetera, indígenas Eperara, Joaquincito, Valle del Cauca, tejeduría en lana, Indígenas Yanaconas, San Sebastián Cauca, tejeduría en fique, indígenas Kankuamos Cesar, tejeduría en fique Arzaria, la Junta y Curazao La Guajira, tejeduría en algodón, indígenas Wayyu, La Guajira tejeduría en algodón, Morroa, Sucre.

145 hojas : ilustraciones, fotografías, 2 disquettes.

Regional Atlántico, San Jacinto, Bolívar. Introducción al manejo del telar horizontal. Regional Centro Oriente, Tipacoque, Boyacá. Tintura con colorantes artificiales tipo disperso, aplicados en trenza o clineja de fique. Curití Santander. Tintura con colorantes naturales aplicados al fique. Investigación de encolantes para hilos y telas en fique, desarrollo de productos.

115 hojas : ilustraciones.

Asesoría de diseño en producción, mejoramiento y desarrollo de nuevos productos para los municipios La Unión, Genóva, San Pablo, Consacá, Sapuyes, Ancuya, Sandoná, La Florida y Linares en los oficios de sombrerería y tejeduría en Iraca (paja toquilla).

132 hojas : ilustraciones a color.

Cuaderno de diseño regional occidente : resguardo indígena de Guangüi, San Miguel de Infi, cestería en paja tetera.

31 hojas.

Asesoría en diseño textil a los artesanos de los municipios de Guateque, Tenza, La capilla, San Mateo y Boavita en el departamento de Boyacá.

[40] hojas : ilustraciones, fotografías.

Asesoría de diseño en los oficios de tejeduría en el municipio de San Agustín, inspecciones Obando y Estrecho, inspección La Jagua del municipio de Garzón y Vereda Belén del municipio La Plata: cestería, trabajos en bambú y cuero en el municipio de San Agustín, sombrería en los municipios de Suaza, Acevedo y Palermo, departamento del Huila.

[60] hojas : ilustraciones, fotografías.

Informe Fase No 2 : Sede: Pasto; Cauca - Municipios del sur: Bolívar, San Sebastián, La Vega, Almaguer.

[158] hojas : ilustraciones, fotografías.