TY - JOUR
T1 - Optimization, validation, and comparison of a rapid method for the quantification of insulin-like growth factor 1 in serum using liquid chromatography–high-resolution mass spectrometry
AU - Seo, Yoondam
AU - Park, Jisoo
AU - Kim, Minyoung
AU - Sung, Changmin
AU - Kwon, Oh Seung
AU - Lee, Hwa Jeong
AU - Min, Hophil
N1 - Publisher Copyright:
© 2020 John Wiley & Sons, Ltd.
PY - 2021/2
Y1 - 2021/2
N2 - Human insulin-like growth factor 1 (IGF-I) is the primary mediator of the effects of the growth hormone (GH). Therefore, it has been used as a biomarker to detect the abuse of GH in sports. The measurement of IGF-I relies on mass-based and immunological approaches to analysis. Among the mass-based analysis methods, liquid chromatography–mass spectrometry (LC–MS) has a number of functional advantages. LC–MS measurements based on the quantification of IGF-I, according to trypsin digestion, are used in the most common method of analyzing doping. However, this method is time-consuming and subject to experimental variability. In this study, we optimized a rapid method for detecting IGF-I without the trypsin digestion step. This method of analysis uses an ultra-centrifugal filter and an LC-HRMS through narrow-range mass scan method. To verify the validity of this method, eight categories of validation testing were applied with the following results: linearity, R2 > 0.99; limit of detection, 15 ng/ml; limit of quantification, 20 ng/ml; accuracy, >99%; recovery rate, >95%; carryover, <0.03; and inter- and intra-day precision values, %CV < 2% and %CV < 6%, respectively. Furthermore, we discussed the correlation of the quantified concentration from two other methods, immunoradiometric assay (IRMA) and parallel reaction monitoring method, using 209 serum samples. In conclusion, although both mass spectrometry-based methods worked equally well in terms of analytical performance and correlation with IRMA results, narrow-range mass scan method had several advantages, such as time and cost savings and reliable reproducibility, over the existing methods.
AB - Human insulin-like growth factor 1 (IGF-I) is the primary mediator of the effects of the growth hormone (GH). Therefore, it has been used as a biomarker to detect the abuse of GH in sports. The measurement of IGF-I relies on mass-based and immunological approaches to analysis. Among the mass-based analysis methods, liquid chromatography–mass spectrometry (LC–MS) has a number of functional advantages. LC–MS measurements based on the quantification of IGF-I, according to trypsin digestion, are used in the most common method of analyzing doping. However, this method is time-consuming and subject to experimental variability. In this study, we optimized a rapid method for detecting IGF-I without the trypsin digestion step. This method of analysis uses an ultra-centrifugal filter and an LC-HRMS through narrow-range mass scan method. To verify the validity of this method, eight categories of validation testing were applied with the following results: linearity, R2 > 0.99; limit of detection, 15 ng/ml; limit of quantification, 20 ng/ml; accuracy, >99%; recovery rate, >95%; carryover, <0.03; and inter- and intra-day precision values, %CV < 2% and %CV < 6%, respectively. Furthermore, we discussed the correlation of the quantified concentration from two other methods, immunoradiometric assay (IRMA) and parallel reaction monitoring method, using 209 serum samples. In conclusion, although both mass spectrometry-based methods worked equally well in terms of analytical performance and correlation with IRMA results, narrow-range mass scan method had several advantages, such as time and cost savings and reliable reproducibility, over the existing methods.
KW - IGF-I
KW - doping control analysis
KW - hGH biomarker
KW - method validation
KW - narrow-range mass scan
UR - http://www.scopus.com/inward/record.url?scp=85093849811&partnerID=8YFLogxK
U2 - 10.1002/dta.2944
DO - 10.1002/dta.2944
M3 - Article
C2 - 33043621
AN - SCOPUS:85093849811
SN - 1942-7603
VL - 13
SP - 451
EP - 459
JO - Drug Testing and Analysis
JF - Drug Testing and Analysis
IS - 2
ER -