Professional Documents
Culture Documents
SECOND EDITION
Edited by
ISBN: 978-0-12-813776-5
Amid Abdullah, MD University of Calgary and Calgary Susan J. Hsiao, MD, PhD Department of Pathology and Cell
Laboratory Services, Calgary, AB, Canada Biology, Columbia University Irving Medical Center,
Maria P. Alfaro, PhD Institute for Genomic Medicine, New York, NY, United States
Nationwide Children’s Hospital, Columbus, OH, Laura M. Jacobsen, MD Department of Pediatrics, Division
United States of Endocrinology, University of Florida, College of
Chris Altomare, BS DRUGSCAN Inc., Horsham, PA, Medicine, Gainesville, FL, United States
United States Kamisha L. Johnson-Davis, PhD Department of Pathology,
Leland Baskin, MD University of Calgary and Calgary University of Utah School of Medicine, ARUP Laboratories,
Laboratory Services, Calgary, AB, Canada Salt Lake City, UT, United States
Lindsay A.L. Bazydlo, PhD Department of Pathology, Steven C. Kazmierczak, PhD Department of Pathology,
University of Virginia, Charlottesville, VA, United States Oregon Health & Science University, Portland, OR,
United States
Jessica M. Boyd, PhD Department of Pathology and
Laboratory Medicine, Cumming School of Medicine, Elaine Lyon, PhD Clinical Services Laboratory,
University of Calgary, Calgary, AB, Canada; Calgary HudsonAlpha Institute for Biotechnology, Huntsville, AL,
Laboratory Services, Calgary, AB, Canada United States
Larry A. Broussard, PhD Department of Clinical Laboratory Gwendolyn A. McMillin, PhD Department of Pathology,
Sciences, Louisiana State University Health Sciences Center, University of Utah School of Medicine, ARUP Laboratories,
New Orleans, LA, United States Salt Lake City, UT, United States
Violeta Chávez, PhD Department of Pathology and Christopher Naugler, MD University of Calgary and
Laboratory Medicine, University of Texas Medical School at Calgary Laboratory Services, Calgary, AB, Canada
Houston, Houston, TX, United States Elena G. Nedelcu, MD Department of Laboratory Medicine,
Alex Chin, PhD University of Calgary and Calgary University of California San Francisco, San Francisco, CA,
Laboratory Services, Calgary, AB, Canada United States
Anthony G. Costantino, PhD DRUGSCAN Inc., Horsham, Andy Nguyen, MD Department of Pathology and
PA, United States Laboratory Medicine, University of Texas McGovern
Medical School, Houston, TX, United States
Amitava Dasgupta, PhD, DABCC Department of Pathology
and Laboratory Medicine, University of Texas McGovern Octavia M. Peck Palmer, PhD Department of Pathology,
Medical School, Houston, TX, United States University of Pittsburgh School of Medicine, Pittsburgh,
PA, United States; Department of Critical Care Medicine,
Pradip Datta, PhD Siemens Healthineers, Newark, DE,
University of Pittsburgh School of Medicine, Pittsburgh, PA,
United States
United States; Department of Clinical and Translational
Robert A. DeSimone, MD Department of Pathology and Science, University of Pittsburgh School, Pittsburgh, PA,
Laboratory Medicine, Weill Cornell Medicine, United States
New York-Presbyterian Hospital, New York, NY,
Amy L. Pyle-Eilola, PhD Pathology and Laboratory
United States
Medicine, Nationwide Children’s Hospital, Columbus, OH,
Uttam Garg, PhD Department of Pathology and Laboratory United States
Medicine, Children’s Mercy Hospitals and Clinics, The
S.M. Hossein Sadrzadeh, PhD Department of Pathology
University of Missouri School of Medicine, Kansas City,
and Laboratory Medicine, Cumming School of Medicine,
MO, United States
University of Calgary, Calgary, AB, Canada; Calgary
Neil S. Harris, MD Department of Pathology, Immunology Laboratory Services, Calgary, AB, Canada
and Laboratory Medicine, University of Florida, College of
Jorge L. Sepulveda, MD, PhD Department of Pathology and
Medicine, Gainesville, FL, United States
Cell Biology, Columbia University Vagelos College of
Joshua Hayden, PhD Department of Pathology and Physicians and Surgeons, New York, NY, United States
Laboratory Medicine, Weill Cornell Medical Center,
New York, NY, United States
xi
xii LIST OF CONTRIBUTORS
Brian Rudolph Shy, MD, PhD Department of Laboratory George Vlad, PhD Department of Pathology & Cell Biology,
Medicine, University of California San Francisco, Columbia University College of Physicians and Surgeons,
San Francisco, CA, United States New York, NY, United States
Aaron Stella, PhD University of Massachusetts Lowell, Amer Wahed, MD Department of Pathology and
Lowell, MA, United States Laboratory Medicine, University of Texas McGovern
Yvette C. Tanhehco, PhD Department of Pathology and Cell Medical School, Houston, TX, United States
Biology, Columbia University Irving Medical Center, William E. Winter, MD Department of Pediatrics, Division
New York-Presbyterian Hospital, New York, NY, of Endocrinology, University of Florida, College of
United States Medicine, Gainesville, FL, United States; Department of
Ashok Tholpady, MD Department of Pathology and Pathology, Immunology and Laboratory Medicine,
Laboratory Medicine, University of Texas MD Anderson University of Florida, College of Medicine, Gainesville, FL,
Cancer Center, Houston, TX, United States United States
Christina Trambas, MD, PhD Chemical Pathologist, Alison Woodworth, PhD Pathology and Laboratory
Chemical Pathology Department, Melbourne Pathology, Medicine, University of Kentucky Medical Center,
Collingwood, VIC, Australia Lexington, KY, United States
Foreword (from the first edition)
Clinicians must make decisions from information showed that when errors were made 75% still produced
presented to them, both by the patient and ancillary results that fell within the reference interval (when
resources available to the physician. Laboratory data perhaps they should not) [1]. Half of the other errors
generally provide quantitative information, which were associated with results that were so absurd that
may be more helpful to physicians than the subjective they were discounted clinically. Such results clearly
information from a patient’s history or physical ex- should not have been released to a physician by the
amination. Indeed, with the prevalent pressure for laboratory and could largely be avoided by a simple
physicians to see more patients in a limited timeframe, review by human or computer before being verified.
laboratory testing has become a more essential compo- However, the remaining 12.5% of errors produced re-
nent of a patient’s diagnostic work-up, partly as a time- sults that could have impacted patient management.
saving measure but also because it does provide The prevalence of errors may be less now than previ-
information against which prior or subsequent test re- ously, since the quality of analytical testing has
sults, and hence patients’ health, may be compared. improved, but the ramifications of each error are not
Tests should be ordered if they could be expected to likely to be less. The consequences of an error vary
provide additional information beyond that obtained depending on the analyte or analytes affected and
from a physician’s first encounter with a patient and if whether the patient involved is an inpatient or outpa-
the results could be expected to influence a patient’s tient. If the patient is an inpatient a physician, if
care. Typically, clinicians use clinical laboratory testing suspicious about the result, will likely have the oppor-
as an adjunct to their history taking and physical tunity to verify the result by repeating the test or other
examination to help confirm a preliminary diagnosis, tests addressing the same physiological functions,
although some testing may establish a diagnosis, for before taking action. However, if the error occurs with a
example molecular tests for inborn errors of metabolism. specimen from an outpatient causing an abnormal result
Microbiological cultures of body fluids may not only to appear normal, that patient may be lost to follow-up
establish the identity of an infecting organism, but also and present later with advanced disease. Despite the
establish the treatment of the associated medical condi- great preponderance of accurate results clinicians should
tion. In outpatient practice clinicians primarily order always be wary of any result that does not seem to fit
tests to assist them in their diagnostic practice, whereas with the patient’s clinical picture. It is, of course, equally
for hospitalized patients, in whom a diagnosis has important for physicians not to dismiss any result that
typically been established, laboratory tests are primarily they do not like as a “laboratory error”. The unexpected
used to monitor a patient’s status and response to result should always prompt an appropriate follow-up.
treatment. Tests of organ function are used to look for The laboratory has a responsibility to ensure that physi-
drug toxicity and the measurement of the circulating cians have confidence in its test results while still
concentrations of drugs with narrow therapeutic win- retaining a healthy skepticism about unexpected results.
dows is done to ensure that optimal drug dosing is Normal laboratory data may provide some assur-
achieved and maintained. The importance of laboratory ance to worried patients who believe that they might
testing is evident when some physicians rely more on have a medical problem, an issue seemingly more
laboratory data than a patient’s own assessment as to prevalent now with the ready accessibility of medical
how he or she feels, opening them to the criticism of information available through computer search engines.
treating the laboratory data rather than the patient. Yet both patients and physicians tend to become over-
In the modern, tightly regulated, clinical laboratory reliant on laboratory information, either not knowing
in a developed country few errors are likely to be made, or ignoring the weakness of laboratory tests, in general.
with the majority labeled as laboratory errors occurring A culture has arisen of physicians and patients
outside the laboratory itself. One study from 1995 believing that the published upper and lower limits of
xiii
xiv FOREWORD (FROM THE FIRST EDITION)
the reference range (or interval) of a test define should be of pursuit of information rather than just
normality. They do not realize that such a range has data. Laboratory information systems provide the po-
probably been derived from 95% of a group of pre- tential to integrate all laboratory data that can then be
sumed healthy individuals, not necessarily selected integrated with clinical and other diagnostic informa-
with respect to all demographic factors or habits that tion by hospital information systems.
were an appropriate comparative reference for a Laboratory actions to highlight values outside the
particular patient. Even if appropriate, 1 in 20 in- reference interval on their comprehensive reports of test
dividuals would be expected to have an abnormal result results to physicians with codes such as “H” or “L” for
for a single test. In the usual situation in which many high and low values exceeding the reference interval
tests are ordered together the probability of abnormal have tended to obscure the actual numerical result and
results in a healthy individual increases in proportion to to cement the concept that the upper and lower reference
the number of tests ordered. Studies have hypothesized limits define normality and that the presence of one of
that the likelihood of all of 20 tests ordered at the same these symbols necessitates further testing. The use of the
time falling within their respective reference intervals is reference limits as published decision limits for national
only 36%. The studies performed to derive the reference programs for renal function, lipid or glucose screening
limits are usually conducted under optimized condi- has again placed a greater burden on the values than
tions such as the time since the volunteer last ate, his or they deserve. Every measurement is subject to analytical
her posture during blood collection and, often the time error, such that repeated determinations will not always
of day. Such idealized conditions are rarely likely to be yield the same result, even under optimal testing con-
attained in an office or hospital practice. ditions. Would it then be more appropriate to make
Factors affecting the usefulness of laboratory data multiple measurements and use an average to establish
may arise in any of the preanalytical, analytical or post- the number to be acted upon by a clinician?
analytical phase of the testing cycle. Failures to consider Much of the opportunity to reduce errors (in the
these factors do constitute errors. If these errors occur broadest sense) rests with the physicians who use test
prior to collection of blood or after results have been results. Over-ordering leads to the possibility of more
produced, while still likely to be labeled as laboratory errors. Inappropriate ordering, for example repetitive
errors because they involve laboratory tests, the labo- ordering of tests whose previous results have been
ratory staffs are typically not liable for them. Yet the normal, or ordering the wrong test or wrong sequence
staff does have the responsibility to educate those in- of tests to elucidate a problem should be minimized by
dividuals who may have caused them to ensure that careful supervision by attending physicians of their
such errors do not recur. If practicing clinicians were trainees involved in the direct management of their
able to use the knowledge that experienced labo- patients. Laboratorians need to be more involved in
ratorians have about the strengths and weaknesses of teaching medical students so that when they become
tests it is likely that much more clinically useful infor- residents their test ordering practices are not learned
mation could be extracted from existing tests. Outside from senior residents who had learned their habits
the laboratory, physicians rarely are knowledgeable from the previous generation of residents. Blanket
about the intra- and interindividual variation observed application of clinical guidelines or test order-sets has
when serial studies are performed on the same in- probably led to much misuse of clinical laboratory
dividuals. For some tests a significant change for an tests. Many clinicians and laboratorians have attemp-
individual may occur when his/her test values shift ted to reduce inappropriate test ordering, but the
from toward one end of the reference interval toward overall conclusion seems to be that education is
the other. Thus a test value does not necessarily have to the most effective means. Unfortunately, the education
exceed the reference limits for it to be abnormal for a needs to be continuously reinforced to have a lasting
given patient. If the preanalytical steps are not stan- effect. The education needs to address the clinical
dardized when repeated testing is done on the same sensitivity of diagnostic tests, the context in which
person, it is more likely that trends in laboratory data they are ordered and their half-lives. Above all edu-
may be missed. There is an onus on everyone involved cation needs to address issues of biological variation
in test ordering and test performance to standardize the and preanalytical factors that may affect test values,
processes to facilitate the maximal extraction of infor- possibly masking trends or making the abnormal
mation from the laboratory data. The combined goal result appear normal and vice versa.
FOREWORD (FROM THE FIRST EDITION) xv
This book provides a comprehensive review of the should be of equal value to clinicians, as to labo-
factors leading to errors in all the areas of clinical lab- ratorians, as they seek the optimal outcome from their
oratory testing. As such it will be of great value to all care of their patients.
laboratory directors and trainees in laboratory medicine
and the technical staff who perform the tests in daily Reference
practice. By clearly identifying problem areas, the book [1] Goldschmidt HMJ, Lent RW. Gross errors and workflow analysis
lays out the opportunities for improvement. This book in the clinical laboratory. Klin Biochem Metab 1995;3:131e49.
Clinical laboratory tests have significant impact on of biotin in the troponin assay. Because people take
patient safety and patient management because more megadoses of biotin, this is a serious public health
than 70% of all medical diagnosis are based on laboratory concern. Therefore, we added a new chapter (Chapter 8).
test results. Physicians rely on hospital laboratories for Another new chapter (Chapter 16) is also added to
obtaining accurate results and a falsely elevated or discuss issues of false negative results in toxicology due
falsely lower value due to interference or pre-analytical to the difficulty in detecting certain drugs such as syn-
errors may have significant influence on diagnosis and thetic cathinone (bath salts) and synthetic cannabinoids
management of patients. Usually, a clinician questions (spices). Chapter 27 is also added to discuss sources of
the validity of a test result if the result does not match errors in flow cytometry. Moreover, Chapters 29e31 are
with clinical evaluation of the patient and calls labora- also newly added chapters in the second edition.
tory professionals for interpretation. However, clini- The objective of this second edition book is to provide
cally significant inaccuracies in laboratory results may a comprehensive guide for laboratory professionals and
go unnoticed and mislead the clinicians into inappro- clinicians regarding sources of errors and misinterpreta-
priate diagnostic and therapeutic approaches, some- tion in the clinical laboratory and how to resolve such
times with very adverse outcomes. The first edition of errors and identify discordant specimens. Accurate lab-
“Accurate Results in the Clinical Laboratory: A Guide oratory result interpretation is essential for patient safety.
to Error Detection and Correction” was published by This book is intended as a practical guide to laboratory
Elsevier in 2013 and was intended as a guide to increase professionals and clinicians who deal with erroneous
awareness of both clinicians and laboratory pro- results on a regular basis. We hope this book will help
fessionals about the various sources of errors in clinical them to be aware of such sources of errors and empower
laboratory tests and what can be done to minimize or them to eliminate such errors when feasible or to account
eliminate such errors. The first edition of the book had for known sources of variability when interpreting
22 chapters and was well received by readers. Due to changes in laboratory results.
success of the first edition, Elsevier requested a second We would like to thank all contributors for taking time
edition of the book. In this edition, we not only updated from their busy professional demands to write chapters.
all chapters of the first edition, but also added 9 new Without their dedicated contributions this project would
chapters so that the second book could be a concise never materialize. We also thank our families for putting
but comprehensive guide for both clinicians and up with us for the last year when we spent many hours
laboratory professionals to detect errors and sources during weekends and evenings writing chapters and
of misinterpretation in the clinical laboratory and to editing this book. Finally our readers will be the judges of
prevent or correct such results. the success of this project. If our readers find this book
Recently, biotin interferences in immunoassays that useful, all the hard work of contributors and editors will
utilize biotinylated antibodies have been described be rewarded.
which may lead to wrong diagnosis of Grave’s disease Respectfully Submitted
due to falsely low TSH (sandwich assay that shows Amitava Dasgupta
negative interference due to biotin) but falsely elevated Houston, TX
T3, T4 and FT4 (competitive immunoassays showing
Jorge L. Sepulveda
positive biotin interferences). The Food and Drug
Administration reported a fatal outcome due to a falsely New York, NY
low troponin value as a result of negative interference
xvii
C H A P T E R
1
Variation, errors, and quality in the
clinical laboratory
Jorge L. Sepulveda
Department of Pathology and Cell Biology, Columbia University Vagelos College of Physicians and Surgeons,
New York, NY, United States
TABLE 1.1 Types of error in the clinical laboratory. TABLE 1.1 Types of error in the clinical laboratory.dcont’d
PRE-ANALYTICAL ANALYTICAL
FIG. 1.1 Example of an error reporting form for the clinical laboratory.
of these approaches is beyond the scope of this book, but TQM approaches apply a system of statistical process
laboratorians and quality management specialists control tools to monitor quality and productivity (quality
should be familiar with these principles for error pre- assurance) and encourage efforts to continuously
vention, error detection, and error management to improve the quality of the products, a concept known
achieve efficient, high-quality laboratory operation and as continuous quality improvement. A major component
patient care [15]. of a quality assurance program is quality control (QC),
which involves the use of periodic measurements of
product quality, thresholds for acceptable performance,
QUALITY IMPROVEMENT IN CLINICAL and rejection of products that do not meet acceptability
LABORATORY criteria. Most notably, QC is applied to all clinical
laboratory testing processes and equipment, including
Quality is defined as all the features of a product that testing reagents, analytical instruments, centrifuges,
meet the requirements of the customers and the health and refrigerators. Typically, for each clinical test,
care system. Many approaches are used to improve external QC materials with known performance, also
and ensure the quality of laboratory operations. The known as controls, are run two or three times daily in
concept of TQM involves a philosophy of excellence parallel with patient specimens. Controls usually have
concerned with all aspects of laboratory operations preassigned analyte concentrations covering important
that impact on the quality of the results. Specifically, medical decision levels, often at low, medium, and
tests, the formula can be modified to take into consider- 95% probability that it is due to the combined analytical
ation the number of repeats in each measurement and intraindividual biological variation; in other words,
(n1 and n2) [27]: the difference between the two creatinine results
sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi (measured without repeats) should exceed 26.8% to be
2 pffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi 95% confident that the change is due to a pathological
RCV95% ¼ 1:96 CVa2 þ CVw2 condition. Conversely, for any change in laboratory
n 1 n2
values, the RCV formula can be used to calculate the
For example, for a serum creatinine measurement probability that it is due to analytical and biological
with an analytical imprecision (CVa) of 7.6% and variation [24,26,27]. See Table 1.2 for examples of RCV
within-subject biologic variation of 5.95%, the RCV at at the 95% confidence limit, using published intraindi-
95% confidence is 26.8% with one measurement for vidual variation and typical laboratory imprecision for
each sample. With two measurements for each sample, each test. Ideally, future LIS should integrate available
the RCV is 18.9%. Therefore, a change between two re- knowledge and patient-specific information and auto-
sults that does not exceed the RCV has a greater than matically provide estimates of expected variation based
TABLE 1.2 Allowable errors and reference change values for selected tests.
Test CVa CVw CVg CLIA TAAE Bio TAAE Allowable imprecision Allowable bias RCV95
All values are percentages. Bio TAAE, total allowable analytical error based on interindividual and intraindividual variation; CLIATAAE, total allowable analytical error
based on Clinical Laboratory Improvement Act (CLIA); CVa, analytical variability in a typical clinical laboratory; CVg, interindividual variability; CVw, intraindividual
qffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
variability. Allowable imprecision ¼ 50% of CVw. Allowable bias ¼ 0:25 CV2w CV2g . RCV95, reference change value at 95% confidence based on CVw and CVa.
Based on Westgard J. Desirable specifications for total error, imprecision, and bias, derived from intra- and inter-individual biologic variation. 2014. Available from: http://www.
westgard.com/biodatabase1.htm.
2
Errors in patient preparation, specimen
collection, anticoagulant and preservative use:
how to avoid such pre-analytical errors
Leland Baskin, Alex Chin, Amid Abdullah, Christopher Naugler
University of Calgary and Calgary Laboratory Services, Calgary, AB, Canada
Cellular elements
Erythrocytes
Leukocytes
Thrombocytes
Proteins Proteins Proteins (excluding fibrinogen)
Patient on therapeutics
Specimen additive
ALT, alanine aminotransferase; AST, aspartate aminotransferase; FAD, flavin adenine dinucleotide; FMN, riboflavin-50 -phosphate; NAD, nicotinamide adenine
dinucleotide; NADP, nicotinamide adenine dinucleotide phosphate; PLP, pyridoxal-50 -phosphate.
a
Plasma or serum folate measurements are usually preferred.
by the hematocrit. Because arteriolar pressure is greater plasma or serum samples. Indeed, there is less water
than that of capillaries and venules, arterial blood will inside erythrocytes compared to the plasma; therefore,
predominate in these samples [2,3]. Given these physio- levels of hydrophilic analytes such as glucose, electro-
logical differences, analytes measured in whole blood do lytes, and water-soluble drugs will be lower in the
not exactly match results obtained from analysis of capillary whole blood [4].
Tube wall Plastic or glass: Plastic preferable Stoppers and stopper lubricants
for safety reasons
Stopper lubricants containing glycerol or silicone
Stopper Inert plasticizers make capping and de-capping of tubes easier, as well
Surfactants Silicone minimizes adsorption of as minimize adherence of cells and clots to stoppers.
analytes, cells Standard red-topped BCT are contaminated with zinc,
Stopper lubricants Ease of capping, de-capping aluminum and magnesium; and all contain varying
amounts of heavy metals [18]. For this reason, most
Clot activator Promote clotting to obtain serum
in plastic collection tubes
labs have specific requirements for collection of speci-
mens for heavy metal assay. Components such as tris
(2-butoxyethyl) phosphate from rubber stopper tubes separating clot results in intracellular leakage of potas-
leaching into the blood during collection have been sium, phosphate, magnesium, and LD into serum,
shown to displace drugs from binding proteins in blood plasma, or whole blood [21]. Ease of use of a single
[19]. Manufacturers have reformulated stopper plasti- centrifugation step, improved specimen analyte stabil-
cizer content to minimize this effect. Triglyceride assays ity, reduced need for aliquoting, convenient storage,
that measure glycerol can be falsely elevated by such and transport in a single primary tube are reasons for
effect. Stopper components and stopper lubricants can preferential usage of SST in the clinical laboratory. It is
also be a potential source of interference with mass very important to follow manufacturer protocols for lab-
spectrometry-based assays [20]. oratory conditions such as proper tube mixing after
collection, centrifugation acceleration and deceleration
speeds, temperature, and storage conditions. Noncom-
Serum separator gel tubes (SST) pliance may result in unexpected degradation of
Separator gels are thixotropic materials that form a separator gel or release of gel components. Gel and
physicochemical barrier after centrifugation in BCT to oil droplets interfere with accuracy and liquid-level
separate packed cells from plasma or serum. Delay in sensing of instrument pipettes, coat cuvettes, and bind
to solid phase in heterogeneous immunoassays. stored at room temperature. EDTA is adequate for
Re-centrifugation, inadequate blood-draw volume, stor- platelet preservation; however, morphological changes
age time, temperature, and drug adsorption may affect occur over time [29]. Clotting can result if there is insuf-
laboratory results. Hydrophobicity of the drug, length ficient EDTA relative to blood. This is usually caused by
of time on separator gel, storage temperature, and meth- overfilling the vacuum tube or poor solubility of EDTA
odology sensitivity are important considerations with (most commonly with disodium salts) [30]. EDTA draws
regard to the stability of therapeutic drugs in BCT. Das- water from cells to artifactually dilute plasma and is
gupta et al. [22] demonstrated that hydrophobic drugs generally not recommended for general chemistry tests.
such as phenytoin, phenobarbital, carbamazepine, quin- EDTA chelates other metallic ions such as copper, zinc,
idine, and lidocaine are adsorbed onto the gel with sig- or magnesium and alters cofactor-dependent activity
nificant decreases (ranging from 5.9 to 64.5%) in serum of many enzymes, such as alkaline phosphatase and cre-
Vacutainer SST tubes. Reformulation of the separator atine kinase, and hence is not used for these chemistry
gel in SST II tubes significantly reduced absorption assays. EDTA is also used for blood bank pretransfusion
and improved performance [23]. Schouwers et al. [24] testing; flow cytometry; Hemoglobin A1c (HbA1c); and
demonstrated minimal effect of separator gel in Star- most common immunosuppressive anti-rejection drugs,
stedt S-Monovette serum tubes for the collection of such as cyclosporine, tacrolimus, sirolimus, and everoli-
four therapeutic drugs (amikacin, vancomycin, valproic mus. Whole blood EDTA in BCT transported on ice is
acid, and acetaminophen) and eight hormones and preferable for the collection of unstable hormones
proteins. susceptible to proteolysis in vitro, such as corticotropin,
parathyroid hormone, C-peptide, vasoactive peptide
(VIP), antidiuretic hormone, carboxy-terminal collagen
Anticoagulants cross-links, calcitonin, renin, procalcitonin, and unstable
Anticoagulants are used to prevent coagulation of peptides such as cytokines. Spray-dried potassium
blood or blood proteins to obtain plasma or whole blood EDTA is the preferred anticoagulant for quantitative
specimens. The most routinely used anticoagulants are proteomic and molecular assay protocols such as
EDTA, heparin (sodium, ammonium, or lithium salts), viral nucleic acid extraction, gene amplification, or
and citrates (trisodium and acid citrate dextrose). Anti- sequencing [27].
coagulants can be powdered, crystallized, solids, or Heparin, a heterogeneous mixture of anionic glycos-
lyophilized liquids. The optimal anticoagulant: blood aminoglycans, inactivates serine proteases in the coagu-
ratio is essential to preserve analytes and prevent clot lation cascadedprimarily thrombin and factors II
or fibrin formation via various differing mechanisms. (prothrombin) and Xadthrough an antithrombin-
In most clinical laboratories, potassium EDTA is the dependent mechanism. For this reason, heparinized
anticoagulant of choice for the complete blood count, plasma is not used for coagulation tests. Typically,
as recommended by the International Council of Stan- lyophilized or solid lithium, sodium, or ammonium
dardization in Hematology [25] and the Clinical and salts of heparin are added to BCT at varying final con-
Laboratory Standards Institute [26]. Dipotassium, tripo- centrations of 10e30 USP units/mL of blood [26]. Hy-
tassium, or disodium salts of EDTA are used as dry or groscopic heparin formulations are used instead of
liquid additives in final concentrations ranging from solutions to avoid dilution effects. Heparin is the recom-
1.5 to 2.2 mg/mL blood when the evacuated BCT is mended anticoagulant for many chemistry tests
filled correctly to its stated draw volume. EDTA acts as requiring whole blood or plasma because chelating
a chelating agent to bind cofactor divalent cations properties and effects on water shifts in cells are mini-
(mainly calcium) to inhibit enzyme reactions in the clot- mal. Heparinized plasma is useful for tests requiring
ting cascadedparticularly the conversion of prothrom- faster turnaround times because it does not require clot-
bin to thrombin and subsequent inhibition of the ting, minimizing the risk of sample pipetting interfer-
thrombolytic action on fibrinogen to fibrin necessary ence due to fibrin microclots [31]. Heparin is the only
for clot formation [27]. For this reason, EDTA plasma anticoagulant recommended for the determination of
is not recommended for coagulation tests such as pro- pH blood gases, electrolytes, and ionized calcium [32].
thrombin time (PT) and activated partial thromboplastin Lithium heparin is commonly used instead of sodium
time (aPTT) [28]. EDTA is an excellent preservative of heparin for general chemistry tests [33]. Obviously, ad-
blood cells and morphology parameters. Stability is ditives may directly affect the measurement of certain
48 h for hgb and 24 h for erythrocytes. Because the hy- analytes. For example, lithium heparin plasma can
pertonic activity by EDTA can alter erythrocytic indices have lithium levels in the toxic range, greater than
and hematocrit, smears should be made within 2 or 3 h 1.0 mmol/L; when filled correctly, sodium heparin tubes
of the blood draw. The white blood cell count remains can elevate sodium levels 1e2 mmol/L; and ammonium
stable for at least 3 days in EDTA anticoagulated blood heparin increases measured ammonia levels [28].
3 Yellow 8e10 times Sodium polyanethol Tube used for mycobacteria (AFB)
sulfonate (SPS) blood culture.
4 Royal blue (with red Not required No additive Tube used for copper and zinc.
band on label)
5 Red glass Not required No additive Tube used for serum tests that
cannot be collected in serum
separator tubes (SST), such as tests
performed by tissue typing. Note:
Red plastic tubes are preferable for
lab tests.
6 Light blue 3e4 times Sodium citrate anticoagulant Tube used mainly for PT (INR),
PTT, and other coagulation studies.
7 Black glass 3e4 times Sodium citrate anticoagulant Tube used for ESR only.
8 Red 5 times Clot activator, and no Tube used for serum tests that
anticoagulant cannot be collected in SST tubes,
such as tests performed by tissue
typing.
9 Gold/Orange 5 times Gel separator and clot Gold top tubes are usually referred
activator to as “SST” (serum separator tube)
and orange top tubes are referred to
“RST” (rapid separator tube). After
centrifugation, the gel forms a
barrier between the clot and the
serum.
10 Dark green Glass (with 8e10 times Sodium heparin anticoagulant Tube used for antimony.
rubber stopper)
11 Dark green 8e10 times Sodium heparin anticoagulant Tube used mainly for amino acids
and cytogenetics tests.
12 Light green (mint) 8e10 times Lithium heparin anticoagulant Usually referred to as “PST”
and gel separator (plasma separator tube). After
centrifugation, the gel forms a
barrier between the blood cells and
the plasma. Tube used mainly for
chemistry tests on acute care
patients.
13 Royal blue (with blue 8e10 times K2EDTA anticoagulant Tube used for trace elements.
band on label)
14 Royal blue (with lavender 8e10 times Na2EDTA anticoagulant Tube used for lead.
band on label)
15 Lavender 8e10 times EDTA anticoagulant Tube used mainly for complete
blood count, pretransfusion testing,
HbA1c, and antirejection drugs.
16 Pale yellow 8e10 times Acid citrate dextrose Tube used for flow cytometry
solution “A” (ACDA) testing.
17 Gray 8e10 times Sodium fluoride and potassium Tube used for lactate.
oxalate anticoagulant
a
Blood collection tubes must be filled in a specific sequence to minimize contamination of sterile specimens, avoid possible test result error caused by carryover of additives between
tubes, and reduce the effect of microclot formation in tubes. When collecting blood samples, allow the tube to fill completely to ensure the blood: additive ratio necessary for accurate
results. Gently invert each tube the required number of times immediately after collection to adequately mix the blood and additive. Never pour blood from one tube into another
tube. See http://www.calgarylabservices.com/files/HealthcareProfessionals/Specimen_Collection/BloodCollectionTubes.pdf for the most current version of this document.
pCO2 regardless of site of collection, but for measure- should not be cultured due to the high risk of contami-
ment of pO2, only blood collected from the earlobe is nation from bacteria growing in the line [2]. Some drugs
recommended [2]. If used for blood gas measurement, such as cyclosporine adhere so tightly to these lines that
care must be taken to minimize exposure to ambient specimens for drug monitoring should always be ob-
air while collecting the specimen [3,47]. A few assays tained from a site unrelated to drug administration [49].
cannot be performed on these specimens, including
erythrocyte sedimentation rate and coagulation studies
and blood cultures [47]. Contamination
Intravenous (IV) fluid is typically composed of water
containing various electrolytes, glucose, and occasion-
Venipuncture ally other substances. Therefore, contamination of a
The median cubital vein in the antecubital fossa is the specimen with this fluid falsely elevates concentrations
most commonly used site due to its accessibility and of these analytes, but at the same time, contamination
size, followed by the neighboring cephalic and basilic causes dilution of the specimen. Thus, values of analytes
veins [2,3,12,48]. Veins on the dorsal surface of the that are not present in the IV fluid should be decreased
hand and wrist, radial aspect of the wrist, followed by [47]. Skin antisepsis is typically accomplished with iso-
dorsal and lateral aspects of the ankle are also used, propanol or iodine compounds [3,48]. Isopropanol is
but these should only be used if one can demonstrate generally recommended. The site should be allowed to
good circulation [3,48]. Sites to be avoided include dry for 30e60 s to minimize the risk of interference
arms ipsilateral to a mastectomy; scarred skin and veins; with alcohol assays. Iodine compounds have been noted
fistulas; sites proximal to (above) intravenous (IV) lines; to affect some assays and probably should be avoided
and edematous, obese, and bruised areas [12]. for chemistry studies. In particular, povidone iodine
can falsely elevate potassium, phosphorous, and uric
acid in specimens collected by skin puncture [3].
Arterial puncture
In order, the radial, brachial, and femoral arteries are
the preferred sites for arterial puncture [2,3,12]. Sites to
Tourniquet effect
be avoided include those that are irritated, edematous, Application of a tourniquet to approximately
and inflamed or infected. Although skin puncture pro- 60 mmHg pressure causes anaerobic metabolism and
vides a similar specimen to arterial blood, for neonates, thus may elevate the lactate and ammonia and lower
specimens for blood gas analysis are best collected from the pH [48]. Tissue destruction may cause the release
an umbilical artery catheter [48]. of intracellular components such as potassium and
enzymes. Venous stasis due to prolonged tourniquet
application (>3 min) may cause significant hemocon-
Indwelling catheters and intravenous lines centration with an 8e10% increase in several enzymes,
For single phlebotomy, it is generally better to avoid proteins, protein-bound substances, and cellular compo-
an area near an IV line [12]. A site in a different extremity nents [3]. Prolongation of venous occlusion from 1 to
or distal to the IV line is preferred. However, for patients 3 min was documented to increase total protein
periodically requiring numerous specimens, collecting (þ4.9%), iron (þ6.7%), lipids (þ4.7%), cholesterol
blood through IV lines and indwelling catheters, (þ5.1%), aspartate aminotransferase (AST) (þ9.3%),
including central venous lines or arterial catheters, and bilirubin (þ8.4%) and to decrease potassium
offers the advantage of facilitating this without phlebot- (þ6.2%) [50]. In addition, stress, hyperventilation, and
omy [2]. This allows staff without extensive phlebotomy muscle contractions (e.g., repeated fist clenching) may
skills to collect blood, thereby freeing experienced elevate analytes such as glucose, cortisol, muscle
phlebotomists to concentrate on other patients. Unfortu- enzymes, potassium, and free fatty acids. For these rea-
nately, this creates an inherent risk for improper spec- sons, it is important to limit venous occlusion to less
imen collection. In order to avoid contamination and than 1 min if possible [48].
dilution with IV fluid; it is recommended that the valve
be closed for at least 3 min prior to specimen collection
[48]. In order to clear the IV fluid from the line, approx-
Hemolysis
imately 6e10 mL should be withdrawn and discarded Hemolysis will elevate the concentration of any con-
[2,48]. Because heparin is often in a line to maintain stituent of erythrocytes and may slightly dilute constitu-
patency, larger volumes may need to be discarded for ents present in low levels in erythrocytes. It becomes
coagulation studies [2]. Blood drawn from these lines significant when the serum concentration of hgb
a perpetual goose.
Procure the heart of a prime ox—the larger the better—hang it up
in a current of dry air as long as it is safe, and at the same time get a
pint of newly-drawn goose oil, which put into a jar along with
Six or eight eschalots, minced
Onions, sliced 1 lb.
Dried sage, powdered 1 oz.
Bay salt ¼ lb.
Saltpetre ½ oz.
Tie brown paper over, and let it remain in a gentle heat until your
meat is ready. First cut out from the heart, the pipe—blood vessel—
as low down as you can, pare away the “deaf ears,” and open as
wide as you consistently can, without piercing the bark or outside
skin, a communication between the two upper cavities—auricles,
and the two lower ones—ventricles, and take out the coagulated
blood. Next rub all parts, the inside and outside, thoroughly twice a
day with the oily mixture for a week, having put the meat, point
downwards, in a straight-sided deep earthen vessel, and keeping the
cavities all the while filled with the liquor. Now boil for fifteen minutes.
Bay leaves, shred 1 oz.
Green laurel, shred 1 oz.
Bay salt, pounded ½ lb.
Vinegar 1 pint
Porter 1 pint
Coarse sugar 1 lb.
Skim it well and add it when half-cold to the meat in the jar, mixing all
well together. Mind that the meat is completely covered with the
pickle, and tie paper over all, so let it be for a week, when boil up all
the pickle, skimming it well, and taking care to renew what may have
been lost or imbibed, and the cavities kept well filled all the time; let it
be in pickle a fortnight longer, then take up, wipe dry inside and out,
make a stuffing of fried sliced onions and sage leaves powdered,
adding black pepper to make it pleasantly hot, and with this fill the
inside of the heart as full as possible, and pressing it in from the top,
make the holes secure with wetted bladder sewed over them. Let it
hang up for a day or two to dry, then wrap it in brown paper and
smoke it, point downwards, for a week; then take it down, rub it for
half-an-hour with olive oil, and smoke it again for a week. This done,
rub it again with the oil and hang it in a quick current of air for twenty-
four hours, and as soon as it is dry enough to retain it, coat it
securely with the gelatine composition, and keep it three months,
and longer the better. Ultimately, it must be roasted, and slices cut
out when cold to be broiled. It is an exceedingly beautiful treat.
the nutriment in fish.
“This is a subject on which I have made some experiments, the
results of which go far to prove that there is much nourishment in fish
—little less than in butcher’s meat, weight for weight; and in effect it
may be more nourishing, considering how, from its softer fibre, fish is
more easily digested. Moreover, there is I find, in fish—in sea-fish—a
substance which does not exist in the flesh of land animals, viz.
iodine, a substance which may have a beneficial effect on the health,
and tend to prevent the production of scrofulous and tubercular
disease—the latter in the form of pulmonary consumption, one of the
most cruel and fatal with which civilised society, and the highly
educated and refined are afflicted. Comparative trials prove that, in
the majority of fish the proportion of solid matter—that is, the matter
which remains after perfect desication or the expulsion of the
aqueous part, is little inferior to that of the several kinds of butcher’s
meat, game, or poultry. And if we give our attention to classes of
people, classed as to the quality of food they principally subsist on,
we find that the ichthyophagous class are especially strong, healthy,
and prolific. In no other class than that of fishers do we see larger
families, handsomer women, or more robust men, or a greater
exemption from the maladies just alluded to.”—Dr. Davy.
collared salmon.
Take a short, thick fish about twelve pounds weight, scale it,
remove the fins, cut off the head with two inches of the jowl, and the
tail with six inches of the fish, these to be cured some other way. Lay
the fish open at the back, take out the bone, wipe nicely and scatter
sifted loaf-sugar over it; after lying six hours replenish the sugar and
leave it till the next day. Next draw your knife down the middle, thus
making two sides of it, which may by cured in different ways. Get a
pint and a half of recently picked shrimps, examine them carefully,
and pound them in a mortar with an anchovy, wiped and boned, and
so much of this mixture as you think sufficient—viz.
Cayenne pepper ½ oz.
Mace, in fine powder ½ oz.
Cloves „ 1 oz.
Bay leaves „ ½ oz.
Table salt 2 oz.
adding a little water that has been boiled. Make a nice smooth paste,
and cover the red surface of the fish with it equally; begin at the head
part, and roll it up into a nice firm collar, which bind tightly with a
broad tape, and sew up in strong calico or light canvas. Let it remain
thus two or three days, then plunge it into a pan of boiling water, with
saltpetre half an ounce, and salt one pound, to each half-gallon of
water; when done enough, take it out, set it on a sieve to cool, and
next day put it in your chimney with a slow fire, to dry gradually, and
then smoke it with
Beech chips 2 parts
Fern 2 parts
Oak lops 2 parts
for a week. When cool take off the cloth, and hang it up in a dry air to
get solid. It may then be enclosed in writing paper and sent to table,
and will be greatly relished. Let the thin side be treated thus: Lay it
down on the skin side, and cover it with rock or bay salt in fine
powder, sifted loaf sugar half a pound, and saltpetre half an ounce;
so let it lie forty-eight hours under a board of tasteless wood,
weighted down. Next wipe it dry, and hang it on your tenterhooks in a
free current of air twenty-four hours; mix well,
Essence of cassia ½ tablespoonful
Essence of cloves 1 tablespoonful
Essence of mace ½ tablespoonful
Essence of cayenne ½ tablespoonful
Essence of bays ½ tablespoonful
lay the fish down on the scaly side, and with a soft flat brush of
camel’s hair, pay it well over with the mixture, and cover with oiled
silk, or its best substitute, to prevent the evaporation of the
essences. Repeat this brushing over three times in twenty-four
hours, and roll it up from the head, binding tightly; expose it to a
current of dry air, and when ready to receive it, give it a fine firm
coating with gelatine composition, and keep it three months in a dry
place. It may be cut in slices for broiling, or if boiled let it be put into
boiling water.
kippered mackerel.
When in season and full of roe, is the time for this process. Take a
dozen mackerel, split them down the back from the head
downwards, and leaving the thin side connected for an inch with the
tail; take out the roes and livers, some of which will be beautiful if
otherwise cured and preserved, remove the gills and refuse, wiping
clean out. Rub the insides lightly with good olive oil, and let them
remain skin side downwards three hours. Boil for a quarter of an
hour the following ingredients, and skim well:
Rock salt or common
salt 1 lb.
Bay salt 1 lb.
Saltpetre ¼ lb.
Coarse sugar 1 lb.
Water 1 gall.
Lay your fish in an earthen pan along with
Thyme 1 handful
Allspice, bruised 1 oz.
Twelve bay leaves, shred
Pour the boiled liquor upon them at about 150 deg. Fahr., and cover
close. In thirty-six hours take out the fish, wipe them dry, stretch
them open by wooden splints at the backs, and hang them in a
strong air current; watch the inside face of them, and if becoming
clammy, place them to a fire for an hour. Smoke them of a nice
chesnut brown colour with
Oak lops or sawdust 2 parts.
Fern or turfs 2 parts.
Beech chips 2 parts.
They will keep well if packed face to face with dry oiled paper
between every two of them. Broil or toast them moderately.
bloaters.
This process is generally conducted in so negligent and rough a
manner—excepting at Yarmouth and Lowestoft—that a little advice
on the subject may not be out of place. As the barrels are emptied of
their contents, the largest fish should be picked out from the rest,
and pickled separately, for otherwise the consumer gets the finest
herrings hardly tasting of salt, and most likely in a state of decay,
while the small ones are so much oversalted, as to be scarcely
eatable. As the fish generally come to hand far from clean, they
should be washed by means of round baskets agitated in tubs of salt
and water, and turned into separate pickling vats, which should have
false bottoms in them, perforated here and there with holes, taps
also being introduced to let off the pickle when required. The safest
and best method is to make use of saturated solutions of salt, which
are made by adding twenty-nine pounds of common salt to seventy-
one pounds of water. The herrings will float in this pickle, but must be
totally immersed by battens of wood laid on the top of them, and held
down by little bags of salt, which, being gradually dissolved, will
maintain the strength of the solution, which is always lessened as
the fish imbibe the muriatic property thereof, and all pickles of this
description are weaker at the surface than at the bottom, and may in
this way be rectified. (See Note, No. 4.) As to the length of time the
fish should remain in the pickle, that depends whether they came to
hand with coarse salt scattered amongst them, at the sea coast, a
precaution necessary in hot weather; a good criterion is when the
fish begin to be stiff or rigid while being handled, but to try one or two
cooked is certainly a sure proof. Pure fresh water must never be
added or made use of in this process after salt has been imbibed, or
the heads will all be broken when putting them on the spits. When
salt enough, run off the brine, and shortly commence putting your
fish on the rods, and hang them up in a current of air, then remove
them to your chimney, and smoke them with
Oak lops 2 parts
Beech chips 2 parts
Fern or grass turfs 2 parts
When they have been smoked enough, return them to the air
currents, as they keep much better on the rods until wanted. If a
constant and full smoke has been kept up, twelve hours will be
sufficient for the smaller fish, and sixteen to eighteen hours for the
large ones. They are not intended to keep good more than four or
five days, but in perfection should be eaten the day after being
cured.
kippered herrings.
The herring is so favourite a fish with the majority of society, that
any improvement in the modes of curing them is a valuable
acquisition. The getting rid of the gut and other objectionable parts
recommends itself, and claims a decided preference over the old
practice of sending the fish to table whole, and, in fact, carrying to
the parlour what ought to have been left in the scullery. The salting
process should be conducted in a similar manner to that for bloaters,
and when taken out of pickle, should be wiped dry, and then split
open at the backs, leaving the bone bare as possible; yet, an inch
from the tail, the thin side should remain attached to the thick side,
this adds much to the appearance of the fish when at table, and
saves the curer some trouble in the succeeding stages of process.
Clean out all the offal and gills, and wipe with cloths dipped in salt
and water, and suspend them by the shoulders upon the tenter
hooks of your rods, thus avoiding the trouble caused by the old plan
of keeping the fish open by splints of wood. Hang them in a free
current of air, and when dried enough—one night is generally
sufficient for that purpose—hang them in the chimney, and smoke
them of a nice chesnut brown colour, and keep them on the rods, but
not in a current, though in a dry room and cold air; when packed it
should be insides faces together, with strips of dry oiled paper
between each two fish.
aberdeen reds.
For this purpose the herrings should be large, full-roed, and fresh.
Immerse them in a pickle of twenty-nine pounds of common salt to
seventy-one pounds of water, and to every pound of salt add half an
ounce of saltpetre. When they become rigid and moderately
flavoured, run off the pickle, put them on the spits, dry them a day or
two, and smoke them with
Oak lops 2 parts
Fern 2 parts
Sawdust 2 parts
until they are of a deep red.
speldings.
At present we are not aware of any superior method of curing the
haddock to the “finnin haddock,” which, if procured soon after they
are drawn from the smoke, are very fine eating. But some seasons
produce these fish in such abundance that it induces curers to save
them by various processes; the small ones may be converted as
follow: Split them open at the belly, right over the backbone, clean
away all the garbage, gills, &c, and lay them in a strong brine of
common salt until nicely flavoured, then hang them on your tenters,
dry them a day or two, taking care they do not become clammy, as
these fish very soon are spoiled. Make a fire in your chimney with
oak lops, sawdust, and beech chips, and when you have brought it
to embers put in the rods, and first dry and then smoke them highly.
Whitings are often done the same way, when the markets are glutted
with the fresh fish.
smoked sprats.
This is a remunerative business when conducted on the best
principles, employing children at trifling wages. I have found the
following to be the best method: Provide a wooden trough eight feet
long by a yard wide, and eighteen inches deep; fix strips of wood an
inch square along the sides, lengthwise of the vat, and six inches
above one another. On these will rest the spits, which must be of iron
wire, a yard long, and so as just to go within the vat. Pick out all the
small fish and rubbish, and wash the bulk in salt and water, as for
bloaters, but not too many at once, as they are apt to sweat if lying
long together, and then would never be bright when smoked. Use a
saturated solution of common salt, or, preferably, of rock salt, and if
you intend to produce “bloated sprats,” two hours will be sufficient to
let them remain in pickle; run off the brine, and put the fish on the
spits, which may be a little pointed at one end. Hang them in a free
current of air till next day, and smoke them with
Oak lops 2 parts
Sawdust 2 parts
Beech or birch chips 2 parts.
until they are the colour of new sovereigns. These will not keep well
more than four or five days, and are generally esteemed. If you want
dried sprats for commerce, let them remain in the brine four hours,
dry them well when on the spits, in a current of air, and when they
begin to lose their plumpness, smoke them with similar fuel till of the
colour of Spanish mahogany. These when packed in boxes, like
cigar boxes, will suit for exportation to the European Continent,
where many thousands of boxes are sent every winter.
aldborough smoked sprats.
Many gentlemen who delight in highly smoked relishes, inquire for
these articles, and as they are seldom to be procured north of the
metropolis, I subjoin an easy way of getting them. In the beginning of
the sprat season—November—take a bushel of fish, pick out all the
largest ones, and with a dozen pounds of common coarse salt or
rock salt at hand, throw a layer of it into the bottom of your salting
tub, then a layer of fish, and so on in alternate layers to the end; let
them lie four hours, mixing them about in the tub two or three times,
this will fix the scales, which are cleared off the fish by the “washing”
process. Now take the sprats up, and with a basket wash them
quickly in very strong salt-and-water, using the same salt if you
choose, and get them on to your spits, and dry them as soon as a
strong current of air will accomplish it. Smoke them with oak alone,
lops and sawdust, until they are of a very dark red colour, and when
quite cold, pack them in round shallow kits, in circles, the heads lying
all one way, and the fish on their backs. The appearance of them is
anything but inviting, yet they are very good, and are always eaten
without cooking. Vast quantities used to be exported to the
Netherlands, Holland, and the German States; they are also well
adapted for sea-stores.
british anchovies.
If it were worth while to favour the deception, you must select your
fish from out of half a bushel of the freshest you can get, retaining
only the middle-sized ones, for the real Gorgona fish are never so
large as our large sprats, and never so small as our little ones, and
your’s should also be all of the same size. Pull off the heads—not
cutting them—in a rough manner, and draw out the gut. Wash not
and wipe not the fish, but put them in straight-sided unglazed
earthen jars, wood is preferable, in layers alternately with this
mixture:
Bay salt 2 lb.
Sal prunelle 2 oz.
Cochineal, in fine 2 oz.
powder
pressing them down as you proceed, and letting the top layer of the
mixture be at least two inches thick. Get cork bungs cut to fit well,
and secure them with plenty of melted resin. Bury the jars in dry
sand in your cellar or store room, “out of the way,” and do not disturb
them for nine months, or till the next sprat season. A fortnight before
you would broach your “prize,” dissolve
Gum dragon 2 oz.
Sal prunelle 2 oz.
Red sanders 1 oz.
in a pint of boiled water, and strain it through flannel, pour it evenly
over the contents of your jars or vessels; secure the bung again, and
in a week or less, turn the receptacles upside-down for a day or two,
and then again set them upright. This is called “feeding” them. And
when all is done, without the aid of “brick-dust,” or what is as bad,
“Armenian Bole,” to give them a fine red colour, the said “British
anchovies” may do to make anchovy sauce of, with other
ingredients, but to bring to table, with dry or buttered toast, as
Gorgona fish—Oh never! See Note, No. 7.
turbot fins.
This idea will naturally suggest itself, that “a pretty expensive
product this will be, by cutting off the fins of a turbot at such a cost;”
but there are fish to be got at much less price that will answer the
purpose, for instance, the brill or brett, and even good firm plaice, in
hard frosty weather, will afford the “amateur” an opportunity of testing
the value of the venture. In a private family, if such a fish came to
table minus its fins it would eat quite as well, even though to the eye
it might not be exactly a handsome dish. Scale the fish, and cut off
the extreme edge of the fins, lay a piece of wood an inch thick on the
body, just to act as a guide to the knife—which must have a very
sharp point—and cut off the fins with an inch and half, or rather
more, of the solid attached; place these upon their bases upright in a