Quality control of Jinzhen Oral Liquid based on amino acids fingerprint and simultaneous determination of 29 amino acids / 中草药

ABSTRACT

Objective: To establish the amino acids fingerprint of Jinzhen Oral Liquid (JOL) and simultaneous determination of 29 amino acids (P-Ser, Tau, Asp, Thr, Ser, Asn, Glu, Gln, Sar, Gly, Ala, Cit, α-ABA, Val, Cys, Ile, Leu, Tyr, Phe, β-Ala, β-AiBA, γ-ABA, Orn, Lys, His, 3-Mehis, Arg, Hypro, Pro) by automatic amino acid analyzer. Methods: The separation was developed on HITACHI HPLC PACKED column (60 mm × 4.6 mm, 3 μm) by gradient elution with citric acid buffer solution at 570 and 440 nm, reaction column temperature was maintained at 115 ℃ and the carrier gas flow rate was 0.35 mL/min and flow rate of derivative pump was 0.30 mL/min. An amino acid fingerprint of JOL was set up, and 16 batches of JOL were evaluated by similarity assay. Furthermore, the contents of the main 29 amino acids were determined. Results: The amino acid fingerprint of JOL was established with good separation, and 29 amino acids were determined simultaneously. The similarity of 16 batches of JOL was about 0.989 to 1.000. Moreover, good linear relationships were found (R2 = 1.000), and the average recovery rates were 95.71%-101.87%, the content range of quantitative components in 16 batches of JOL was as following P-Ser 15.457 2-29.362 4 μg/mL, Tau 64.423 4-114.238 6 μg/mL, Asp 19.056 4-32.549 0 μg/mL, Thr 6.704 8-7.841 8 μg/mL, Ser 22.609 4-39.382 8 μg/mL, Asn 61.134 0-115.456 0 μg/mL, Glu 32.254 6-63.127 2 μg/mL, Gln 5.223 8-8.953 2 μg/mL, Sar 4.081 0-44.007 6 μg/mL, Gly 12.403 0-23.516 4 μg/mL, Ala 33.876 8-44.257 4 μg/mL, Cit 3.514 2-9.881 0 μg/mL, α-ABA 1.126 4-2.287 8 μg/mL, Val 11.584 6-15.469 0 μg/mL, Cys 1.660 2-4.041 8 μg/mL, Ile 4.087 8-5.469 2 μg/mL, Leu 8.295 2-11.724 0 μg/mL, Tyr 7.492 6-10.761 2 μg/mL, Phe 4.856 4-9.248 0 μg/mL, β-Ala 2.309 4-6.782 6 μg/mL, β-AiBA 0.175 0-14.790 6 μg/mL, γ-ABA 17.654 4-26.621 8 μg/mL, Orn 42.338 4-58.172 6 μg/mL, Lys 12.994 9-28.498 0 μg/mL, His 1.802 6-4.067 4 μg/mL, 3-Mehis 2.608 4-4.384 0 μg/mL, Arg 107.141 6-301.649 8 μg/mL, Hypro 15.811 6-37.807 6 μg/mL, Pro 143.714 6-243.956 6 μg/mL. Conclusion: The method for the determination and analysis of amino acids in JOL by automatic amino acid analyzer has the advantages of good reproducibility, reliable results, and the qualitative and quantitative study of amino acids in JOL has clarified the contribution of Caprae hircus cornu's horn to the composition of preparation to a certain extent, which provides a useful supplement for the construction of the quality evaluation system of JOL.