Analysis of seven folates in food by LC–MS/MS to improve accuracy of total folate data

An LC–MS/MS method for analyzing seven folates in food was developed and validated. 5-Methyltetrahydrofolate, 5-formyltetrahydrofolate, 10-formylfolic acid, tetrahydrofolate and folic acid were quantified using a stable isotope dilution assay (SIDA) with deuterated analogues as internal standards. Additionally, 10-formyldihydrofolate and 5,10-methenyltetrahydrofolate were quantified using deuterated internal standards different in structure. Due to interconversion of 5,10-methenyltetrahydrofolate to 5-formyltetrahydrofolate and 10-formyldihydrofolate to 10-formylfolic acid during sample preparation, a SIDA was not considered because of a resulting double calculation of the amounts interconverting. [²H₄]-5-methyltetrahydrofolate was used as internal standard for 5,10-methenyltetrahydrofolate, due to a similar retention time, and [²H₄]-10-formylfolic acid as well as [²H₄]-5-methyltetrahydrofolate was used for 10-formyldihydrofolate, because no internal standards co-elute. To confirm that no matrix effects affect the quantitation of 5,10-methenyltetrahydrofolate and 10-formyldihydrofolate, postcolumn infusion experiments were performed. Validation of the assay was accomplished by determining linearity, precision, recovery, limit of detection and limit of quantitation. The latter parameters were partly obtained by application of a dual-isotope label design including [¹³C₅]-labeled folates. The amounts of 5,10-methenyltetrahydrofolate in the purified extracts of different food samples ranged between 0.3 and 1.3 % and for 10-HCO-H₂folate between 0.05 and 8 % of the total folate amount. Correction for incomplete recovery of the latter folate during cleanup indicates even higher contents. Therefore, especially 10-formyldihydrofolate should not be neglected to obtain accurate results for folates.     

Frequency conversion of Ti:sapphire-based femtosecond laser systemsto the 200-nm spectral region using nonlinear optical crystals

We review the linear and nonlinear optical properties of crystals transparent near and below 200 nm and suitable for up conversion of femtosecond Ti:sapphire laser sources, β-BaB₂O₄ , the crystal with the largest birefringence of all presently available materials, is investigated experimentally as a quadrupler by mixing the fundamental and the third harmonic both using a 1-kHz repetition rate Ti:sapphire regenerative amplifier and a 82-MHz mode-locked Ti:sapphire laser. Milliwatt average powers near 200 nm are achieved in both cases. The sub-200-fs pulses at the fourth harmonic are almost bandwidth limited. Sum-frequency generation as a method for upconversion of femtosecond pulses is experimentally studied by mixing the fourth harmonic generated down to 189 nm by the regenerative amplifier with a parametrically generated femtosecond pulse in the infrared. Pulse energies at the microjoule level are produced with LiB ₃O₅ above 180 nm. Li₂B₄O₇ shows superior performance in the 170-180-nm range, and the shortest wavelength achieved with KB₅ O₈·4H₂O is 166 nm