Biomedical Engineering Reference
In-Depth Information
development of standard methods and reference materials for the determination of
hormones in human serum [
84-
86
]. A number of researchers also proposed a series
of standardization procedures, including method development, validation, data
interpretation and correlation between different bioanalytical techniques, and estab-
lishing international databases of human hormones [
2,
14,
18
]. In contrast to the
clinical laboratories, EPA considers steroid hormones as environmental pollutants
in water and sediments, and it has established an LC-MS/MS Method #539 follow-
ing EPA Chemical QC Guidelines [
77
] .
4.1
Guidelines for GLP, Reference Standards,
and Analytical Methodology
Current Good Laboratory Practice from Food and Drug Administration (FDA GLP,
21CFR58) is a general guideline for pharmaceutical industry, and Good Laboratory
Practice from Environmental Protection Agency (EPA GLP, 40CFR792) is a guide-
line for agriculture chemical industry, and environmental protection agencies and
organizations. Both guidelines emphasize on compliance in personnel, facilities,
articles, and documentation for animal and analytical experiments. In order to stan-
dardize the analytical methodology of steroid hormone analysis, the clinical and the
environmental laboratories and facilities should follow FDA or EPA GLP guide-
lines. These guidelines include the following major requirements: (1) to train related
analysts, (2) to calibrate and maintain instruments, (3) to characterize reference
standard, (4) to conduct analytical testing following standard operation procedures
(SOPs), (5) to record analytical procedures, protocols, reports, deviations, investiga-
tion, etc. Besides GLPs, guidelines for reference standards and analytical methodol-
ogy are also important for steroid hormone analyses, as summarized in Table
5
.
4.2
Reference Standards
The primary reference standards, including internal standards, of steroid hormones
used for LC-MS/MS or GC-MS analyses in GLP laboratories should be obtained
from authentic sources, e.g., US Pharmacopeia (USP) and NIST. Relative inexpen-
sive secondary or working standards for daily testing may be used as alternatives of
the primary standards, and they may be obtained from commercial sources. These
working standards should be characterized against the primary reference standards
using compendia or validated methods before being applied for GLP testing pur-
poses. A reference standard program should be established in each laboratory, insti-
tute, or company to monitor the specification, quality, characterization, stability,
storage, inventory, and replacement of those reference standards. A certificate of
analysis should be issued after characterization of each standard batch, and it should
include the information of manufacturer or source, date of manufacture, date of
