FRANKLIN CRUISES FR 8/90, 5/92 AND 8/93
DATA DOCUMENTATION
JGOFS WESTERN EQUATORIAL PACIFIC PROCESS STUDY

[1] General:

Parameter:	Phytoplankton pigments detemined by HPLC: Cruises  FR 9008 and FR 9205.	
Level 1	Yes
Principal Investigator:	Harry Higgins
Institute Address:		CSIRO Division of Marine Research
E-Mail Address:		Harry.Higgins@marine.csiro.au

List of Parameters:	Phytoplankton pigments:	
                                
Cli a		chlorophyllide a
Chl c3		chlorophyll c3
Chl c1+c2		chlorophyll c1+c2
PERI		peridinin
cPERI		cis-peridinin
19BUT		19’-butanoyloxyfucoxanthin
FUCO		fucoxanthin
c19BUT		cis-19’-butanoyloxyfucoxanthin
NEO		neoxanthin
19HEX		19’-hexanoyloxyfucoxanthin
cFUCO		cis-fucoxanthin
c19HEX		cis-19’-hexanoyloxyfucoxanthin
PRAS		prasinoxanthin
Pb a		pheophorbide a
VIOL		violaxanthin
Pb a like 1		pheophorbide a like 1
DINO		dinoxanthin
cPRAS		cis-prasinoxanthin
Pb a like 2		pheophorbide a like 2
DDX		diadinoxanthin
DDC		diadinochrome
ANTH		antheraxanthin
ALLO		alloxanthin
MON		monadoxanthin
DIAT		diatoxanthin
LUT		lutein
ZEA		zeaxanthin
cZEA		cis-zeaxanthin
CANT		canthaxanthin
SIPN		siphonein
Chl b + dvChl b		chlorophyll b + divinyl-chlorophyll b
Chl a'		chlorophyll a allomer
Chl a + dvChl a		chlorophyll a + divinyl-chlorophyll a
eChl a		chlorophyll a epimer
ECHN		echinenone
Ph b		pheophytin b
Ph a		pheophytin a
ßYCA		ßY-carotene
eeCA		ee-carotene
ßeCA		ße-carotene
ßßCA		ßß-carotene
pPh a		pyro-pheophytin a
cßeCA		cis-ße-carotene
cßßCA		cis-ßß-carotene

List of Units:			g m-3  

[2] Sampling:

Gear (e.g. CTD, pump, etc.):	CTD; 10 litre niskin bottles
Standard Depths:	Hydrochemistry depths: see Hydrochemistry data
Chemicals used:			none
Special Procedures:	Niskins with silicone rubber o-rings and closure rubbers. Began pressure filtration through Whatman GFF filers as soon as CTD on deck. Filters blotted dry and stored in liquid nitrogen until analysed.
Comments and Notes:	Sampled in dim light.

[3] Analysis:

Instrument:			HPLC 
Method:	extraction of pigments from filters followed by ternary gradient HPLC 
Precision:	coefficient of variation estimated as 17% over all pigments for triplicate samples from FR 9205
Comments:			(1) FR 9205: 
•	Duplicate extract from station 42, 75.1 m
•	Duplicate extract from station 51, 96 m 
•	Triplicate samples from station 45, 67 m
•	147 0E transect and 155 0E, 3 0S samples not worked up
(2) FR 9308:
Due to loss of all pigment samples on FR 9308, chlorophyll a data can be estimated using fluorescence profiles from FR 9308 and fluorometer calibration from FR 9205

[4] Results:

Quality of Data:	FR 9008 and FR 9205: good.  FR 9308: loss of HPLC pigment samples; chlorophyll a can be calculated from fluorometric data (calibrated using FR 5/92 data; see methods for a full description of the calibration procedures).
Known Problems:	Loss of pigment samples for FR 8/93.  

 [5] Brief description of analytical methods

Chl a estimation from in situ fluorescence

During FR05/92, the fluorometer was calibrated against measurements of extracted Chl a (actually chlorophyll a plus divinyl‑chlorophyll a ‑ see Mackey et al., 1995) determined by HPLC with diode array detection. The relationship was:
 Chl a (g l-1)  =  0.01204  x  Seatech(%)  +  0.026       (r2 = 0.698, n = 94)
with a standard error in Chl a of 0.06 g l-1. During FR08/90, the instrument was calibrated against Chl a determined spectrophotometrically and the correlation was:
Chl a (g l-1)  =  0.01239  x  Seatech(%)  +  0.0142       (r2 = 0.848, n = 174)
with a standard error in Chl a of 0.05 g l-1. Between the two cruises, the slope had changed by only 3% and the difference in intercept was less than 20% of the standard error in the calculated concentration of Chl a. Unfortunately, samples collected for calibration of the fluorometer on FR08/93 had decomposed because of a faulty Dewar before they could be analysed. We therefore assumed that the calibration for FR08/93 was unchanged from that found in 1992. 
Reference:  see Mackey et al., Deep-Sea Research, 44, 1951-1978.

Pigment calibration
Water samples (10 L) were generally collected at 25 m intervals to 150 m from Niskin bottles attached to the CTD rosette. The sampling depth closest to the DCM, determined from the in situ fluorescence profile, was moved so that samples were always collected from the DCM. The samples were pressure‑filtered (5 psi) through Whatman GF/F filters which were blotted dry and stored in liquid nitrogen. In contrast to FR08/90 where pigment analyses of samples was carried out post-cruise in Hobart within 1 - 3 months of collection, pigment samples from FR05/92 were analysed on-board within 24 hours of sampling. 
The pigment filters were extracted with acetone (Carpenter et al., 1991) and analysed by HPLC based on the ternary gradient method of Wright et al., (1991) as described in Mackey et al., (1995). Pigments were detected at 436 nm and identified by their retention time and spectra. Calibration standards of Chl a1 and Chl b1, and carotenoids from the SCOR-recommended algal cultures (Jeffrey and Wright, 1997) were kindly made available by S. W. Wright. HPLC response factors were determined by the method of external standards (Mantoura and Repeta, 1997) using data provided by (Jeffrey, et al., 1997) and S. W. Wright  (personal communication). 

Reference:  see Higgins, H. W. and Mackey, D. J. (2000)  Deep-Sea Research, 47, 1461-1483.

References:

Carpenter, P.D., Butler, E.C.V., Higgins, H.W., Mackey, D.J., and Nichols, P.D. (1991)  Chemistry of trace elelments, humic substances, and sedimentary organic matter in Macquarie Harbour, Tasmania.  Australian Journal of Marine and Freshwater Research 42, 625-654.

Higgins, H. W. and Mackey, D. J. (2000) Algal class abundances, estimated from chlorophyll and carotenoid pigments, in the western Equatorial Pacific under El Niño and non-El Niño conditions. Deep-Sea Research, 47, 1461-1483.

Jeffrey, S.W. and Wright, S.W. (1997).  Qualitative and quantitative analysis of SCOR reference algal cultures.  In:  Jeffrey, S.W., Mantoura, R.F.C., Wright, S.W. (Eds.), Phytoplankton pigments in Oceanography: Guidelines to Modern Methods.  SCOR-UNESCO Paris, pp 343-360. 

Mackey, D. J., Higgins, H. W., Mackey, M. D. and Holdsworth, D. (1998) Algal class abundances in the western equatorial Pacific: estimation from HPLC measurements of chloroplast pigments using CHEMTAX. Deep-Sea Research, 45, 1441-1468.

Mackey, D. J., Parslow, J. S., Griffiths, F. B., Higgins, H. W. and Tilbrook, B. (1997) Phytoplankton productivity and the carbon cycle in the western equatorial Pacific under ENSO and non-ENSO conditions. Deep-Sea Research, 44, 1951-1978.

Mantoura, R.F.C. and Repeta, D.J. (1997)  Calibration methods for HPLC. In:  Jeffrey, S.W., Mantoura, R.F.C., Wright, S.W. (Eds.), Phytoplankton pigments in Oceanography: Guidelines to Modern Methods.  SCOR-UNESCO Paris, pp 407-428.

Wright, S.W., Jeffrey, S.W., Mantoura, R.F.C., Llewellyn, C.A., Bjornland, T., Repta, D., Welschmeyer, N. (1991) Improved HPLC mehtod for the analysis of chlorophylls and carotenoids from marine phytoplankton.  Marine Ecology Progress Series 77, 183-196.

[6] Comments: