Proced ures

2. Proced ures

All experimental i nformation is entered into a bound research notebook with a black waterproof i n k pen . Any mistakes i n the notes are marked out with a single stroke through the erro r. Corrections are written beside or above the m istake. N otes should be written neatly so that others can read them. It is suggested that

i n structi ons be written in fi rst person , active voice. For example: "I placed the TLC plate i nto the chamber at 3 :00 P.M. "

1 ) Copy tem plate i nto research notebook prior to analysis

I n order to note all the i m portant details of the experiment while it is in progress, it is helpful to copy the following template into the research

notebook prior to analysis.

2) Record the following i nformation as the analysis proceeds:

a. Date

b. Name of tech nician

c. Type of TLC analysis

d. Pu rpose of analysis ( 1 -2 sentences)

e. Type of chamber and approxi m ate dimensions

f. Type of development ( i . e . , sandwich chamber or conventional; si ngle or m u ltiple)

g. Elu ent system (with man ufacturer and lot of solvents u sed)

h. Type of TLC plate (with man ufacturer and lot used)

i. Preparation of TLC plate ( i . e . , prewashed and activated or as received) j. Start time of analysis k . Stop time of analysis

I. Total time m. D istance traveled by eluent

n. Detection reagent (with manufacturer and lot used , and date prepared)

3) Prepare a table and record the fol lowing:

Written Documentation of the TLC Plate 123

d . Date solution was prepared (and notebook page number)

e. N u mber of separation zones

i. distances

i i . Rf values

i i i . notes

4) Make a photocopy of the TLC plate After t h e detection a n d eval uation o f t h e T L C plate, make a photocopy o f the

plate and adhere it to the notebook with an archival ad hesive. Note: Do not use tape. An acid-free wh ite glue is recom mended for attaching notes, ph oto­ copies of plates , etc . , to the notebook.

Tem p l ate for Written Documentation of a TLC Plate Date :

Name:

Type of analysis:

Pu rpose:

Cham ber: Type of development: Eluent syste m : TLC plate :

Preparation of plate : Start time:

Stop time:

Total ti me: Distance traveled : Detection reagent:

Lane Su bstance DistancelRf values Notes

1.

2.

3.

4.

5.

6.

7.

8.

9.

1 0.

11 .

12.

13.

14.

15.

1 6.

Protocol F

Photodocumentation of the TLC Plate Using Visible Light

Sum mary Guideli nes for photographic documentation of a TLC plate

Equipment:

35-mm camera Copy stand

Light box (similar to those used to view slides) Two tungsten lamps Light meter (optional)

Film:

Black-and-white fil m: 35-mm Kodak TMAX 1 00 Color slide film: 35-mm Kodak Ektachrome P rofessional

1 60T (tungsten balanced)

Time: Approximately 30 minutes per plate (less in a large series)

Scope Ph otography is u sed to q u ickly and accurately record the TLC results u nder either normal or ultraviolet light. This protocol descri bes the proced u res used to ph oto­

graph a TLC plate u nder normal l ight. For photodocumentation of TLC plates u nder ultraviolet light, see Protocol G .

Photography is the preferred method for permanently record ing t h e results of a TLC experi ment. TLC separation zones can be detected with reagents that

create visible spots, such as those seen for amino acids when detected with n i n hyd ri n . Such a TLC plate i s photographed using tungsten photoflood lamps (balanced 3200 OK) fo r i l l u m ination . The TLC plate is ph otographed with black-and -wh ite print fi lm and color slide fi l m under normal i l l u m i nation using

a 3 5 - m m camera on a copy stand .

Striegel . Hill

Scheme Documentation of Thi n-Layer Chromatography by