University of Massachusetts, Lowell
College of Health Sciences
LABORATORY METHODS OF NUTRITION ASSESSMENT
SYLLABUS (Course #36.465-565)
WEED HALL, ROOM 205
Mon: 5 PM-8 PM
FALL, 2013
Instructor: Professor Handelman
Office: Room 308, Weed Hall
Email: Garry_Handelman@uml.edu
Phone: 978-934-4503
Office hours:
Mon, 1-2:30 PM, Wed, 3-4:30 PM,
or by appointment
INTRODUCTION AND COURSE OBJECTIVES:
The objectives of this course are to understand how to carry out and interpret laboratory methods
of nutritional assessment. Emphasis will primarily be on chromatographic methods, and other
laboratory methods will be examined as well. Nutritional status can often be determined by
measurement of bloodstream levels of a nutrient. However, for every nutrient, dietary intake is
only of several factors that affect plasma levels, and determination of nutritional status often
requires consideration of several different biological components.
Further indicators for nutritional status are often obtained by physical examination (skin lesions,
failure to achieve height for age, injury, cardiac impairment, poor dark adaptation, neurological
symptoms), or from dietary history. Many of the symptoms associated with nutritional deficiency
can arise from other causes, and definitive diagnosis often requires multiple procedures. For
example, rickets can be caused by vitamin D deficiency, but there are genetic causes as well,
which must be considered in the diagnosis.
From this overview, the student should appreciate that lab measurements of nutrients are useful,
but in many cases other information is needed, to reach a valid assessment of nutritional status
and its influence on health outcomes. In this course, we will learn some of the lab procedures
that can be used for nutritional assessment, and discuss their interpretation in the clinical and
public health setting.
Nutrients and related markers that will be assessed, or discussed, in plasma and whole
blood:
-iron (Hb, Zn-EPP, serum iron, ferritin): emphasis on iron status and iron deficiency
-vitamin C, by HPLC with electrochemical detection
-vitamin E and A, by HPLC with uv-vis detection
-vitamin D, by liquid chromatography/mass spec (LC-MS)
Each chromatographic assay uses a DIFFERENT method for detecting the molecules. We
will discuss in detail how the measurement takes advantage of a property of the molecule
that allows it to measured by the lab apparatus.
This class will teach you quantitative techniques of pipetting and sample measurement, and
provide experience with HPLC, mass spec, and UV-VIS spectrophotometry. The analytical
principles provided by this course should enable you to tackle a wide range of nutritional
analyses.
Laboratory protocols will be provided for each procedure, and supplemented with additional
materials throughout the semester. Critical literature reviews will aid in learning how to
interpret the clinical and public health importance of laboratory measures of nutrition status.
This will develop your ability to evaluate the meaning of nutrition measurements.
Throughout the course, there will be emphasis on the importance of each step in the
protocol. You should be able to explain what the purpose of each step is, toward the
objective of measuring the nutrient. During class reviews, groups of students will use the
white board to explain the entire protocol to the class. You will have the opportunity to
practice with the steps in each protocol, before you run the actual sample.
All course materials will be posted at the website provided for my classes:
Health.uml.edu/Handelmannutrition. In addition, I may Email you class notices and
supplemental materials.
VALUE OF LABORATORY ASSESSMENT OF NUTRITIONAL STATUS:
SELECTION OF METHODS THAT PROVIDE USEFUL INFORMATION
The decision to make a laboratory measurement must include the likelihood that the
measurement will provide useful information. In many cases, factors in addition to dietary
intake have a major influence. The measurement of Hb allows us to know if the person is
anemic, but anemia arises from several causes, and iron deficiency is only one cause. A
physician who diagnoses anemia has the responsibility to consider several potential causes
of for the anemia. Plasma vitamin A levels can be low from deficiency, but can also
decrease during inflammation. Plasma zinc has limited relation to dietary intake, and is
controlled mostly by other biological factors. Most people regulate plasma sodium over a
very narrow range, despite a broad variation in dietary sodium intake, and your daily sodium
consumption cannot be estimated from your plasma sodium level.
At the other extreme, vitamin C levels usually correspond very well to the amount in the diet,
and levels of plasma omega-3 fatty acids are strongly linked to dietary intake of this class of
fatty acids.
In the case of extreme nutritional deficiency, as seen in clinical practice, levels of most
nutrients in the plasma are a useful diagnostic indicator, but this is a special case, since
healthy people rarely show these very depressed levels.
OVERALL PLAN OF THE COURSE
You will conduct a variety of analyses on your own blood sample, or on another sample that
will be provided if you choose. You will work with a group of students on each lab
procedure. This course will require strict adherence to safety procedures because of
potential biohazards from blood samples. These safety protocols will be reviewed in detail
on the first day of class. We will have a lab safety video at the first class meeting.
You should make every effort to achieve 100% attendance, since this will help everyone
progress as a group through these procedures. The practical experience gained in the lab
is not readily obtained by reading a book on these methods.
The due dates for the lab writeups are indicated on the class schedule. I will distribute a sample
writeup, to illustrate the procedures to be followed in writing up your results. There are 6
writeups, and each writeup is worth 10 points toward your grade.
We will meet several times in Weed 205, but will also use Weed 309 and other labs for
some meetings of the class.
For part of the semester, the class will be organized into separate groups: as will be
explained on the first day of class. Each group will perform all the laboratory procedures for
the entire course.
GRADING
The graded assignments are indicated on the class schedule. There are 150 points
possible in this course. There will be 1 exam (50 points), 2 quizzes (20 points each), and
6 lab reports (10 points each).
ASSIGNMENT
DATE
POINTS
QUIZ #1
Oct 16
20
Lab reports, set 1
Oct 21
30
QUIZ #2
Nov 25
20
Lab reports, set 2
Dec 2
30
Final exam
Dec 9
50
TOTAL POINTS
150
Students enrolled at the 500 level will have an additional assignment,
which will be discussed.
Grades are assigned on the following scale:
A =90-100; A- =87-89; B+ =84-86; B =80-83;B- =77-79; C+ =74-76;
C =70-73; C- =67-69, D+ =64-66; D =60-63; F = anything less than 60.
LAB SAFETY
Because we will be handling blood samples and chemical reagents, there are rigorous safety
rules that must be followed, for everyone’s protection. These are described in the handout that
was distributed. Essential rules for safety: lab coats, long pants, closed shoes, gloves, and face
shields.
The rules will be described in detail at the first lecture. We will have a safety video the first day
of class.
ACADEMIC POLICY: HONESTY ON EXAMS AND CLASS ASSIGNMENTS
Cheating on an examination will result in loss of all the credit for that exam
toward your grade. If there is a second occurrence of cheating, you will receive a
mandatory grade of F for the entire course.
For written assignments, materials must be prepared without use of plagiarism.
The handout on academic writing describes this policy in detail.
SCHOOL OF HEALTH AND ENVIRONMENT: SOCIAL MEDIA POLICY
The School of Health and Environment recognizes that all involved in health care have
a moral, ethical and legal responsibility to maintain individual’s rights to privacy. HIPAA
protects patient privacy by law and includes any individually identifiable patient
information in oral or recorded form where the information could identify an individual by
name, medical condition, demographic data or other means.
Students in the School of Health are expected to act with honesty, integrity and
respect the privacy rights of others. All students in the School of Health and Environment
are expected to meet their professional responsibilities when using social media and other
electronic networks including but not limited to blogs, instant messaging, social
networking sites, email, public media sites and photographs. This policy prohibits posting
written material or photographs that identify patients, health care agencies, educational
institutions or other students in clinical sites or patient related activities. This policy
applies whether using University devices and computers or personal equipment. In
addition, all School of Health and Environment students are required to abide by clinical
agency policies related to the use of social media and technological resources.
Failure to adhere to this policy may result in probation, suspension or dismissal from
the School of Health and Environment and/or legal prosecution under the requirements of
HIPAA.
USE OF CELL PHONES, IPADS, AND OTHER ELECTRONIC MEDIA
If use of portable electronic devices during class becomes a distraction or interferes
with the learning process, I will require students to stop using these devices in class.
While we are in the classroom, we give our attention fully to materials that are being
considered for the course.
COURSE OBJECTIVES
1)
For human clinical disorders, be able to consider how nutritional assessment
might contribute to diagnosis and management
2)
Understand secondary factors (in addition to nutrition) that change the tissue level
of a nutrient, or a nutrition status marker
3)
Develop technical skills in sample processing for nutrition measurements,
including HPLC and ELISA.
4)
Consider the chemical features of different nutrients that allow their measurement
by analytical techniques
5)
Understand the basic principles of chromatographic separation
6)
Review the basic information (and limitations) that can be gained from
laboratory measurement of nutrition status
Class Schedule: Laboratory Methods of Nutritional Assessment
Sept 9
Introductory class: discussion of class objectives and class structure.
Review of the strengths and limitations of lab methods.
Discussion of hazards, and video on lab safety.
Review the plan for blood sample collections.
Practice with use of pipettes, to develop good technique.
Lecture on iron status and assessment: Hb, ZnEPP and other methods.
Sept 16
Donate blood sample, run basic iron measures (Hb, ZnEPP).
Aliquot plasma samples for measurements later in semester.
Assigned readings distributed for class on Sept 23.
Sept 23
Assessment of iron status from a blood sample: techniques and discussion
Overview of the problem of iron deficiency, and how it can be assessed.
Review of iron methods: Hb, serum iron, Zn-EPP, ferritin.
Practice the steps in the ferritin ELISA protocol.
Sept 30
Run the ELISA protocol for ferritin.
Oct 7
Review all data collected on iron status.
Student-led discussion of the ferritin protocol.
Review for quiz on Oct 16.
Oct 16
Quiz: Lab Methods of Iron Assessment (20 points).
Introduction: theory of HPLC methods of assessment.
Demonstration of each HPLC method.
Oct 21
Practice with steps in all HPLC protocols..
Make schedule for HPLC analyses
Lab reports due: Pipette lab, Hb and Zn-EPP, ferritin (30 points).
Oct 28
Group 1: Vitamin E+A, vitamin D
Nov 4
Continue with HPLC analyses in groups.
Nov 11
Veterans Day: university closed, class not held.
Nov 18
Group 2: Vitamin C
\
Discussion, entire class: progress on HPLC protocols
Continue with HPLC protocols in groups.
Review for quiz: sample preparation for chromatography.
Nov 25
Quiz: sample preparation for chromatography (20 points).
Complete all HPLC assays.
Dec 2
LAB REPORTS DUE (30 points): VITAMIN MEASUREMENTS
Student-led review on HPLC methods: practical aspects.
STRENGTHS AND LIMITATIONS OF LAB METHODS OF ASSESSMENT
OF NUTRITION STATUS.
Review for final exam
Dec 9
Final exam: HPLC in nutrition assessment (50 points)