Quantitative analysis of triglycerides using atmospheric pressure chemical ionization‐mass spectrometry

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Abstract
Atmospheric pressure chemical ionization‐mass spectrometry (APCI‐MS) was used for quantitative analysis of triglycerides (TG) separated by reverse‐phase high‐performance liquid chromatography. APCI‐MS was used for analysis of mono‐acid TG standards containing deuterated internal standard, of a synthetic mixture of heterogeneous TG, of randomized and normal soybean oils and of randomized and normal lard samples. Quantitation of the TG by four approaches based on APCI‐MS were compared, and these were compared to quantitation obtained using liquid chromatography (LC) with flameionization detection (FID). The APCI‐MS methods were based on (i) calibration curves from data for mono‐acid TG standards, (ii) response factors obtained from a synthetic mixture of TG, (iii) response factors calculated from comparison of randomized samples to their statistically expected compositions, and (iv) response factors calculated from comparison of fatty acid (FA) compositions calculated from TG compositions to FA compositions obtained by calibrated gas chromatography (GC) with FID. Response factors derived from a synthetic mixture were not widely applicable to samples of disparate composition. The TG compositions obtained using APCI‐MS data without application of response factors had average relative errors very similar to those obtained using LC‐FID. Numerous TG species were identified using LC/APCI‐MS which were undetected using LC‐FID. Two quantitation methods, based on response factors calculated from randomized samples or on response factors calculated from FA compositions, both gave similar results for all samples. The TG compositions obtained using response factors calculated from FA compositions showed less average relative error than was obtained from LC‐FID data, and were in good agreement with predicted compositions for the synthetic mixture and for randomized soybean oil and lard samples.