Allureâ¢ Acidix HPLC Column - Chromtech.com.au

Restek Advantage, Summer 1999: Allure[TM] Acidix HPLC Column Specialized for LC/MS Analysis of Acids and Amino Acids 

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Allure Acidix 

HPLC Column 

Specialized for LC/MS 

Analysis of Acids and 

Amino Acids 

Summer 1999 

derivatization 

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Improved LC/MS sensitivity 

Maximum retention of acidic compounds 

High selectivity and reproducibility 

Analysis of amino acids without 

Restek's Allure phases are engineered to provide maximum sensitivity for liquid 

chromatography/mass spectrometry (LC/MS) separations. The Allure Acidix 

phase retains and exhibits good peak shape for compounds that contain 

carboxylic acid, sulfonic acid, phosphoric acid, or other acidic functional groups. 

Maximum retention occurs at high percentages of organic modifier, which results 

in a desirable increase in the signal-to-noise ratio for LC/MS analyses. 

Use of the Allure Acidix column allows chromatographers to generate highthroughput 

acid and amino acid analyses by eliminating the need for derivatization 

of these analytes. Typically, amino acids require derivatization prior to separation 

by HPLC. This adds much cost and time to the method, and should be avoided 

whenever possible. In addition, incomplete derivatization can distort quantitative 

measurements. This is a problem particularly when more than one moiety on an 

amino acid can be derivatized. Until now, underivatized amino acids only could be 

run using ion exchange, but many of the buffers required in these methods are not 

LC/MS compatible. The Allure Acidix column retains and separates amino acids, 

achieving good peak shape using simple isocratic mobile phases (Figure 1). 

Volatile buffers and high organic content can be used in the mobile phase to assist 

in the LC/MS ionization process for good MS sensitivity.

Restek Advantage, Summer 1999: Allure[TM] Acidix HPLC Column Specialized for LC/MS Analysis of Acids and Amino Acids 

However, most of the advantages exhibited by the Allure Acidix are not limited 

to LC/MS separations. Fast analyses, excellent peak shape and retention, high 

reproducibility, and the elimination of the need for derivatization apply to other 

HPLC systems as well. Polar acids (e.g., ascorbic acid) that are difficult to retain 

by traditional C18 columns are easily retained by the Allure Acidix phase (Figure 

2). Larger molecular weight acidic compounds such as pharmaceuticals also show 

excellent retention and peak shape (Figure 3). Additionally, salicylic acid and 

aspirin are cleanly and easily separated on the Allure Acidix column (Figure 4). 

The Allure Acidix column is a breakthrough in HPLC stationary phase 

development that permits the analysis of amino acids without derivatization. 

Furthermore, it provides the highest sensitivity available for LC/MS detection of 

acidic compounds, and it retains most acids that C18 phases cannot. For samples 

with solubility limitations, the Allure Acidix column can be used at both the high 

and the low end of percent organic in the mobile phase. Most importantly, it 

displays the reproducibility and reliability chromatographers have come to expect 

from Restek chromatography products. 

Product Listing: 

Particle Size: 5µm 1.0mm ID 2.1mm ID 3.2mm ID 4.6mm ID 

30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9162531 

9162551 

9162511 

9162561 

9162521 

9162571 

9162532 

9162552 

9162512 

9162562 

9162522 

9162572 

9162533 

9162553 

9162513 

9162563 

9162523 

9162573 

9162535 

9162555 

9162515 

9162565 

9162525 

9162575

Restek Advantage, Summer 1999: Integra-Guard[TM] Columns -- Get the Protection Without the Connection! 

Summer 1999 

Integra-Guard Columns -- Get the 

Protection Without the Connection! 

Some people swear by press-fit connectors, and others swear at 

them. For many analysts, the art of attaching a guard column to an 

analytical column is a mystery. Restek's chemists have discovered 

the solution to this mystery -- the most reliable connection is no 

connection at all! No guard column system is more permanent than 

one continuous length of tubing containing both the guard column 

and the analytical column. 

Restek pioneered this innovative approach to guard columns, and our Integra-Guard 

column is engineered to be worry-free for the analyst. The transition area between the guard 

and analytical column is the point at which the guard column ends and the analytical column 

begins. High-temperature string is used to tie the guard column in a separate loop, making it 

easy to distinguish from the analytical column. The entire setup is suspended in Restek's 

unique “crush-free” cage, which prevents the column from coming in contact with anything 

that could damage it. We offer Integra-Guard columns in a wide variety of phases (Table I). 

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Guard columns are beneficial because they: 

Increase the lifetime of a column by trapping non-volatile contamination on the first few 

meters 

Allow for system maintenance without sacrificing separation 

Function as "retention gaps," allowing components with different volatilities to be 

focused in the same band on the analytical column 

Problems that conventional guard columns can create: 

Difficult and time consuming to make a proper connection 

Leaks may develop during an analysis, destroying the columns 

Active sites and dead volume from tubing connectors may cause peak tailing and loss 

of resolution 

Integra-Guard Columns provide the solution: 

The most reliable connection between a guard column 

and an analytical column is pictured at the right: NO 

CONNECTION AT ALL 

Integra-guard technology eliminates the need for a 

connector by incorporating the guard column into the 

actual analytical column

Restek Advantage, Summer 1999: Integra-Guard[TM] Columns -- Get the Protection Without the Connection! 

Table I: Phases currently available with an 

Integra-Guard guard column 

Integra-Guard Guard Columns* 

Rtx®-1 

Rtx®-1MS 

Rtx®-5 

Rtx®-5MS 

XTI®-5 

Rtx®-1301 

Rtx®-624 

Rtx®-1701 

Rtx®-Volatiles 

Rtx®-20 

Rtx®-35 

Rtx®-BAC 1 

Rtx®-BAC 2 

Stabilwax® 

Stabilwax®-DA 

Stabilwax®-DB 

ID length suffix # 

0.25mm 5m -124 

10m -127 

0.32mm 

0.53mm 

5m 

10m 

5m 

10m 

-125 

-128 

-126 

-129 

*Add suffix # and price to the order 

of your column. 

If you are currently using a guard column or considering one for the future, call Restek today 

and ask about our Integra-Guard columns.

Restek -- LC/MS Columns 

Restek HPLC Columns 

Ultra IBD Specialized Columns for LC/MS 

Physical Characteristics: 

Particle Size: 5µm spherical 

Pore Size: 100 Å 

Average Carbon Load: 12% 

Chromatographic Properties: 

Not end-capped 

pH Range: 2.5 to 7.5 

Temperature Limit: 80°C 

This packing is in the class of intrinsically base-deactivated (IBD) phases, containing a polar 

group within, or intrinsic to, the hydrocarbon bonded phase. 

Applications: 

The polar group assists in deactivating surface silanols and participates in providing unique 

separation selectivities of acids, bases, and zwitterionics, as well as other polar compounds. 

more about Ultra IBD 

Ultra IBD 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

9175331 

9175351 

9175311 

9175361 

9175321 

with Trident inlet fitting: 

30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

9175332 

9175352 

9175312 

9175362 

9175322 

9175332-700 

9175352-700 

9175312-700 

9175362-700 

9175322-700 

9175333 

9175353 

9175313 

9175363 

9175323 

9175333-700 

9175353-700 

9175313-700 

9175363-700 

9175323-700 

9175335 

9175355 

9175315 

9175365 

9175325 

9175335-700 

9175355-700 

9175315-700 

9175365-700 

9175325-700 

Particle Size: 5µm 1.0mm ID 2.1mm ID 3.2mm ID 4.6mm ID


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9175531 

9175551 

9175511 

9175561 

9175521 

9175571 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

Ultra IBD 

Guard Cartridges: 

9175532 

9175552 

9175512 

9175562 

9175522 

9175572 

9175532-700 

9175552-700 

9175512-700 

9175562-700 

9175522-700 

9175572-700 

Dimensions 

10 x 2.1mm 

10 x 4.0mm 

20 x 4.0mm 

9175533 

9175553 

9175513 

9175563 

9175523 

9175573 

9175533-700 

9175553-700 

9175513-700 

9175563-700 

9175523-700 

9175573-700 

cat.# 

917550212 

917550210 

917550220 

9175535 

9175555 

9175515 

9175565 

9175525 

9175575 

9175535-700 

9175555-700 

9175515-700 

9175565-700 

9175525-700 

9175575-700 

Qty. 

3 

3 

2

Restek -- Ultra IBD Intrinsically Base Deactivated HPLC Column 

Ultra IBD Intrinsically Base Deactivated 

HPLC Column 

formerly Pinnacle IBD 

I. Introduction II. Applications III. Summary 

IV. Product Listing 

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Intrinsically base deactivated for excellent peak shape and selectivity of acids, bases, 

zwitterions and polar compounds 

Exceptional peak shape even under neutral mobile phase conditions 

Bonded on base deactivated Type B Silica 

Highly reproducible and stable 

Retains polar analytes 

Immune to bonded phase chain collapse 

Alternative to ion pairing method development 

HPLC Columns 

See Also: NERCDG '98 Presentation 

I. Introduction 

Intrinsically base deactivated (IBD) phases are an exciting new class of reversed-phase packings that 

contain a polar group within, or intrinsic to, the hydrocarbon bonded chain. In contrast to conventional 

C18 phases, IBD packings retain solutes not only by hydrophobic interactions, but also by polar 

attractive forces between the stationary phase and the analyte. 

The Ultra IBD phase was designed to address some of the shortcomings associated with conventional 

C18 packings. First, the Ultra IBD phase retains polar solutes that are not well retained on C18 columns. 

This is possible because IBD phases contain a polar group, not available on C18 phases, that can attract 

polar analytes. Second, the Ultra IBD phase furnishes unique selectivity compared to C18 columns. 

Again, this is due to the polar group of the IBD structure. Third, the Ultra IBD phase does not collapse, 

http://www.restekcorp.com/pinn_ibd.htm (1 of 3) [12/7/01 8:56:21 PM]


as do most C18 chains under high aqueous mobile phase conditions (see Figure 1). Chain folding on C18 

phases results in complete or partial loss of retention and resolution. Fourth, the Ultra IBD phase shows 

excellent peak shape for acids, bases, and zwitterions. The polar group plays an active role in 

deactivating undesirable silanol interactions with the analyte. As a result, samples containing mixtures of 

widely different pKa's can be run simultaneously. Finally, the Ultra IBD phase is often a good choice for 

LC-MS applications. Simple, MS-compatible mobile phases can be used with IBD columns while 

maintaining good peak shape and selectivity. IBD columns are an excellent alternative to ion pair 

methods on conventional C18 phases. 

II. Applications 

Figure 1A depicts the chain collapsing phenomenon associated with most C18 and some C8 phases under 

high aqueous mobile phase conditions. With insufficient organic content in the mobile phase, the 

hydrophobic chains of the phase fold down on themselves to obtain an energetically more favorable 

position, to avoid the outer aqueous, hydrophilic environment. The chains are no longer in a position to 

interact with the analytes, resulting in partial or complete loss of retention, depending on the degree of 

chain folding. In contrast, under the same high aqueous mobile phase conditions, the Ultra IBD phase 

provides essentially no net gain in energy from chain folding, because the polar group within the chain is 

hydrophilic. Therefore, the chains remain extended and can continue to interact with the analytes (Figure 

1B). Typically, this scenario is chromatographically manifested in reversed-phase mode as plotted in 

Figure 1C, where retention continues on an upward swing (no loss of retention) even at 100% aqueous 

mobile phase composition. 

Peak shape of basic compounds on silica bonded phases can be improved by lowering the pH to 

protonate active silanols. However, there are times when more neutral conditions are required, such as 

with samples that are labile at acidic pH's. The Ultra IBD phase performs well even at neutral pH (see 

Figure 2), which makes it a particularly valuable packing in this instance. 

Figures 3 and 4 serve as examples of how the Ultra IBD phase performs well for acid/base mixtures. 

Many C18 packings show good peak shape for acids (in the case of active phases) or bases (in the case of 

deactivated phases), but not for both. The Ultra IBD phase exhibits good peak shape for a wide variety of 

solute types, at both acidic (Figure 3) and neutral (Figure 4) pH. 

It is noteworthy that the Ultra IBD phase is similar to conventional C18 phases in one aspect. That is, 

acidic mobile phases decrease retention for basic analytes and increase retention for acidic analytes. To 

increase retention for bases and/or increase retention for acids, use a neutral pH mobile phase. 

Retention of polar compounds that are difficult to obtain on C18 phases can often be retained on the 

Ultra IBD phase. The famotidine/ranitidine chromatogram in Figure 5 is indicative of this increased 

retention of polar compounds. 

III. Summary 

Ultra IBD columns are intrinsically base deactivated phases for reversed-phase separations of acids, 

bases, zwitterions and polar compounds. They demonstrate strength in areas weakly serviced by 

http://www.restekcorp.com/pinn_ibd.htm (2 of 3) [12/7/01 8:56:21 PM]


traditional C18 columns, including retention of polar solutes, good peak shape for compounds containing 

a wide variety of pKa's, and the ability to use high aqueous mobile phases in separations. Employment of 

Ultra IBD columns in place of conventional C18 phases often eliminates the need for ion pair reagents in 

the mobile phase. This is especially important for LC/MS work. Generally, they also are more selective 

than C18 phases. 

IV. Product Listing 

Ultra IBD Columns, 5µm, 100Å 

Cat # 

Description 

91755 Bulk Silica 

9175532 

30 x 2mm 

9175552 

50 x 2mm 

9175512 

100 x 2mm 

9175562 

150 x 2mm 

9175522 

200 x 2mm 

9175572 

250 x 2mm 

9175533 

9175553 

9175513 

9175563 

9175523 

9175573 

9175535 

9175555 

9175515 

9175565 

9175525 

9175575 

917550212 

917550210 

917550220 

30 x 3mm 

50 x 3mm 

100 x 3mm 

150 x 3mm 

200 x 3mm 

250 x 3mm 

30 x 4.6mm 

50 x 4.6mm 

100 x 4.6mm 

150 x 4.6mm 

200 x 4.6mm 

250 x 4.6mm 

10 x 2mm (3-pk.) 

10 x 4mm (3-pk.) 

20 x 4mm (2-pk.)



Allure Basix Specialized Columns for LC/MS 



Fully end-capped 






Highly retentive phase for analytes containing amino group functionality. 

Applications: 

Ideal LC/MS column for basic solutes. Excellent for pharmaceutical and other amine-containing 

compound separations. 

more about Allure Basix 

Allure Basix 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9161531 

9161551 

9161511 

9161561 

9161521 

9161571 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

Allure Basix 


9161532 

9161552 

9161512 

9161562 

9161522 

9161572 

9161532-700 

9161552-700 

9161512-700 

9161562-700 

9161522-700 

9161572-700 

Dimensions 

10 x 2.1mm 

10 x 4.0mm 

20 x 4.0mm 

9161533 

9161553 

9161513 

9161563 

9161523 

9161573 

9161533-700 

9161553-700 

9161513-700 

9161563-700 

9161523-700 

9161573-700 

cat.# 

916150212 

916150210 

916150220 

9161535 

9161555 

9161515 

9161565 

9161525 

9161575 

9161535-700 

9161555-700 

9161515-700 

9161565-700 

9161525-700 

9161575-700 

Qty. 

3 

3 

2 

http://www.restekcorp.com/basix.htm (1 of 2) [12/7/01 8:56:41 PM]

Restek -- LC/MS Columns

RAS[TM] -- Allure[TM] Basix HPLC Column 

NEW Allure Basix HPLC Column 

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Maximum retention of basic (amino-containing) compounds 

Increased LC/MS sensitivity 

Excellent general purpose column 

Highly reproducible and stable 


High Performance Liquid Chromatography/Mass Spectrometry (LC/MS) has developed into one of the 

most powerful analytical techniques available. Although the mass spectrometer can, in itself, provide 

enhanced selectivity, quality chromatographic separations are essential in order to garner the full 

advantages of the technique. Among the advantages of LC/MS technology is an increased level of 

sensitivity compared to that provided by traditional ultraviolet (UV) detection. The sensitivity achieved 

in the LC/MS system is related to the efficiency of the ionization/evaporation process. A greater 

percentage of organic modifier in the mobile phase improves the ionization/evaporation efficiency and 

thus provides increased sensitivity. 

The Allure family of HPLC columns has been designed to provide the retention and resolution 

required for successful LC/MS method development. Specifically, the Allure Basix phase has been 

engineered to produce maximum retention of basic (amino-containing) compounds using mobile phases 

with high organic percentages. For LC/MS applications, this unique ability translates into increased 

sensitivity for the target analytes. Therefore, the Allure Basix phase is an excellent column choice for 

pharmaceutical applications ranging from raw material analysis to metabolic studies. 

APPLICATIONS 

Figure 1 depicts the increased retention of the basic analyte amitriptyline at high organic percentages, as 

compared to the Allure C18 phase. At lower percentages of organic, hydrophobic interactions prevail 

and the C18 phase provides greater retention. As shown in Figure 2, the increase in retention at high 

organic content invokes polar interactions as the dominant separation mechanism, and produces 

significantly enhanced signal-to-noise (S/N) in the mass spectrometer -- nearly a 250% increase! 

Figure 1: Allure Basix Column Provides Increased Retention of Basic Compounds at 

http://www.restekcorp.com/basix/basix.htm (1 of 3) [12/7/01 8:56:47 PM]


High Organic Mobile Phase 

Figure 3 demonstrates that the Allure Basix column is also an excellent choice for difficult as well as 

routine analyses. The series of tricyclic antidepressants exhibit excellent peak shape and are well 

resolved using a simple mobile phase. 

Application: Corticosteroids on Allure Basix 

SUMMARY 

The Allure Basix column is an ideal LC/MS column for the analysis of basic analytes. The column is 

reproducible, stable, and an excellent choice for routine analysis of basic compounds. 

Allure Basix HPLC Column 

Specialized for Basic Compounds 

Allure Basix Columns, 

5µm, 60Å 2.1mm ID 3.2mm ID 4.6mm ID 

http://www.restekcorp.com/basix/basix.htm (2 of 3) [12/7/01 8:56:47 PM]


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9161532 

9161552 

9161512 

9161562 

9161522 

9161572 

Allure Basix Guard Cartridges 

10 x 2mm (3-pk.) 916150212 

10 x 4mm (3-pk.) 916150210 

20 x 4mm (2-pk.) 916150220 

9161533 

9161553 

9161513 

9161563 

9161523 

9161573 

9161535 

9161555 

9161515 

9161565 

9161525 

9161575 

Bulk Silica 

91615



Allure PFP Propyl Specialized Columns for 

LC/MS 









An excellent phase for separating nucleosides, nucleotides, purines, pyrimidines, halogenated 

compounds, ß-blockers, and tricyclic antidepressants. 

Applications: 

Nucleosides, nucleotides, purines, pyrimidines, and halogenated compounds. 

Allure PFP Propyl 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9169531 

9169551 

9169511 

9169561 

9169521 

9169571 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 



9169532 

9169552 

9169512 

9169562 

9169522 

9169572 

9169532-700 

9169552-700 

9169512-700 

9169562-700 

9169522-700 

9169572-700 

Dimensions 

10 x 2.1mm 

10 x 4.0mm 

20 x 4.0mm 

9169533 

9169553 

9169513 

9169563 

9169523 

9169573 

9169533-700 

9169553-700 

9169513-700 

9169563-700 

9169523-700 

9169573-700 

cat.# 

916950212 

916950210 

916950220 

9169535 

9169555 

9169515 

9169565 

9169525 

9169575 

9169535-700 

9169555-700 

9169515-700 

9169565-700 

9169525-700 

9169575-700 

Qty. 

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http://www.restekcorp.com/pfp_prop.htm (1 of 2) [12/7/01 8:57:08 PM]

Restek -- LC/MS Columns



Ultra C18 USP L1 Columns 



Pore Size: 100Å 



End-capped 



A very retentive, high-purity reverse phase packing that exhibits excellent peak shape for a wide 

range of compounds. 

Applications: 

Carbonyls, fatty acids, glycerides, anilines, fat-soluble vitamins, phthalates, barbiturates, 

steroids, PTH amino acids, and acids. 

Ultra C18 (USP L1) 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

9174331 

9174351 

9174311 

9174361 

9174321 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

9174332 

9174352 

9174312 

9174362 

9174322 

9174332-700 

9174352-700 

9174312-700 

9174362-700 

9174322-700 

9174333 

9174353 

9174313 

9174363 

9174323 

9174333-700 

9174353-700 

9174313-700 

9174363-700 

9174323-700 

9174335 

9174355 

9174315 

9174365 

9174325 

9174335-700 

9174355-700 

9174315-700 

9174365-700 

9174325-700 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9174531 

9174551 

9174511 

9174561 

9174521 

9174571 

9174532 

9174552 

9174512 

9174562 

9174522 

9174572 

9174533 

9174553 

9174513 

9174563 

9174523 

9174573 

9174535 

9174555 

9174515 

9174565 

9174525 

9174575



30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

Ultra C18 


9174532-700 

9174552-700 

9174512-700 

9174562-700 

9174522-700 

9174572-700 

Dimensions 

10 x 2.1mm 

10 x 4.0mm 

20 x 4.0mm 

9174533-700 

9174553-700 

9174513-700 

9174563-700 

9174523-700 

9174573-700 

cat.# 

917450212 

917450210 

917450220 

9174535-700 

9174555-700 

9174515-700 

9174565-700 

9174525-700 

9174575-700 

Qty. 

3 

3 

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Allure C18 USP L1 Specialized Columns for 

LC/MS 









Highly retentive phase for hydrophobic and slightly polar analytes. Mobile phases containing 

higher percentages of organic modifier contribute to greater sensitivity in LC/MS. 

Applications: 

Ideal LC/MS column for neutral to slightly polar solutes. Analyzes basic compounds, showing 

good deactivation for basic solutes; excellent for explosives and steroid separations. 

more about Allure C18 

Allure C18 (USP L1) 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9164531 

9164551 

9164511 

9164561 

9164521 

9164571 


30mm length 

50mm length 

100mm length 

150mm length 

200mm length 

250mm length 

9164532 

9164552 

9164512 

9164562 

9164522 

9164572 

9164532-700 

9164552-700 

9164512-700 

9164562-700 

9164522-700 

9164572-700 

9164533 

9164553 

9164513 

9164563 

9164523 

9164573 

9164533-700 

9164553-700 

9164513-700 

9164563-700 

9164523-700 

9164573-700 

9164535 

9164555 

9164515 

9164565 

9164525 

9164575 

9164535-700 

9164555-700 

9164515-700 

9164565-700 

9164525-700 

9164575-700 

http://www.restekcorp.com/allurc18.htm (1 of 2) [12/7/01 8:58:40 PM]


Allure C18 


Dimensions 

10 x 2.1mm 

10 x 4.0mm 

20 x 4.0mm 

cat.# 

916450212 

916450210 

916450220 

Qty. 

3 

3 

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Restek Advantage, Fall 2000: Ultra Aqueous C18 HPLC Column -- Reversed-phase analysis of polar compounds 

- RESTEK HOME - LIBRARY - RELATED PRODUCTS 

Fall 2000 

Ultra Aqueous C18 HPLC Column 

Reversed-phase analysis of polar compounds 

by Terry Reid, HPLC R&D Chemist 

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● 

● 

Permits use of highly aqueous mobile phases 

Enhanced retention and selectivity for polar analytes 

Excellent base deactivation 

Highly polar compounds such as short-chain organic acids, nucleotides, 

catecholamines, and water-soluble vitamins are difficult to retain using 

conventional reversed phase columns, even with little or no organic solvent in the 

mobile phase. The Ultra Aqueous C18 column enhances the retention of polar 

analytes, while maintaining a high level of base deactivation and a selectivity that 

is similar to a conventional C18 column. Six organic acids that are difficult to retain 

using many conventional C18 columns can be well retained on the Ultra Aqueous 

C18 column (Figure 1). 

The unique qualities of the Ultra Aqueous C18 column are apparent when 

comparing this column to the Ultra C18 column -- a highly retentive, welldeactivated, 

general-purpose C18 column -- in a separation of polar compounds. 

The Ultra Aqueous C18 column has very similar retention and selectivity to the 

Ultra C18 column when analyzing neutral, hydrophobic compounds. While phenol, 

a neutral polar compound, is retained similarly by both columns (Figure 2), the 

basic compound pyridine is retained approximately 2.5 times longer on the Ultra 

Aqueous C18 column. Also note that, in spite of its greater retention, pyridine 

elutes from the Ultra Aqueous C18 column as a relatively symmetrical peak 

(pyridine is commonly used as a test probe for column base deactivation). This 

proves that enhanced retention for polar compounds is achieved on the Ultra 

Aqueous C18 column without sacrificing base deactivation. 

The Ultra Aqueous C18 column is designed using Type B, high-purity silica, and a 

novel bonding chemistry that produces a true C18 phase (USP L1) with alkyl 

chains that remain completely extended, even when continually exposed to a 

highly aqueous mobile phase. This is because polar groups on the silica surface


keep the stationary phase wetted. The unique secondary polar character prevents 

chain folding (i.e., the hydrophobic C18 chains do not self-associate or "fold down" 

onto the silica surface to avoid associating with a very hydrophilic mobile phase), 

and enhances the retention and selectivity of polar compounds without 

compromising the level of base deactivation. Ultimately, this means that the Ultra 

Aqueous C18 column offers stable and reproducible retention, even with 100% 

aqueous mobile phases. 

Product Listing 

Ultra Aqueous C18 HPLC Columns 

5µm particle size 

length 1.0mm ID 2.1mm ID 3.2mm ID 4.6mm ID 

30mm 

50mm 

100mm 

150mm 

200mm 

250mm 

9178531 

9178551 

9178511 

9178561 

9178521 

9178571 

with Trident inlet fitting 

30mm 

50mm 

100mm 

150mm 

200mm 

250mm 

3µm particle size 

9178532 

9178552 

9178512 

9178562 

9178522 

9178572 

9178532-700 

9178552-700 

9178512-700 

9178562-700 

9178522-700 

9178572-700 

9178533 

9178553 

9178513 

9178563 

9178523 

9178573 

9178533-700 

9178553-700 

9178513-700 

9178563-700 

9178523-700 

9178573-700 

9178535 

9178555 

9178515 

9178565 

9178525 

9178575 

9178535-700 

9178555-700 

9178515-700 

9178565-700 

9178525-700 

9178575-700 

length 1.0mm ID 2.1mm ID 3.2mm ID 4.6mm ID 

30mm 

50mm 

100mm 

150mm 

200mm 

250mm 

9178331 

9178351 

9178311 

9178361 

9178321 

9178371 

with Trident inlet fitting 

9178332 

9178352 

9178312 

9178362 

9178322 

9178372 

9178333 

9178353 

9178313 

9178363 

9178323 

9178373 

9178335 

9178355 

9178315 

9178365 

9178325 

9178375 

http://www.restekcorp.com/advntage/d00nine.htm (2 of 3) [12/7/01 8:58:49 PM]


30mm 

50mm 

100mm 

150mm 

200mm 

250mm 

9178332-700 

9178352-700 

9178312-700 

9178362-700 

9178322-700 

9178372-700 

9178333-700 

9178353-700 

9178313-700 

9178363-700 

9178323-700 

9178373-700 

9178335-700 

9178355-700 

9178315-700 

9178365-700 

9178325-700 

9178375-700

Restek -- HPLC Methods Development & Validation Kits 

< Restek HPLC Columns 

HPLC and LC/MS Column Kits 

Simplify HPLC and LC/MS 

Methods Development and Validation 

Methods Development and 

Validation Kits 

The HPLC chromatographers 

at Restek understand the 

unique problems encountered 

in pharmaceutical 

laboratories. Chemists from 

our HPLC applications group 

have worked in 

pharmaceutical laboratories 

and are experienced with 

methods development and 

validation. Because of this 

insight, Restek has introduced 

LC/MS and HPLC 

development and validation 

kits containing a wide variety 

of columns to handle virtually 

any application. The selection 

of the right column reduces 

the need for special modifiers 

to coax the stationary phase 

into providing retention. 

Simple, yet rugged methods 

that improve reproducibility, 

enhance sensitivity, and 

Feature 

Variety of columns 

Quality assurance 

Convenient ordering 

and packaging; kits 

tailored to fit your 

needs 

Contains columns 

for normal or 

reverse phase modes 

Benefit 

Able to handle virtually 

any pharmaceutical 

application 

Columns meet stringent 

requirements, thereby 

increasing reproducibility 

Quick and uncomplicated 

help with method 

development 

Optimizes separations of 

acids, bases, anions, 

cations, and weakly 

charged analytes from 

matrix components 

As competition increases in the 

pharmaceutical industry, the ability to create 

quick, rugged, accurate, and precise high 

performance liquid chromatography (HPLC) 

and liquid chromatography/mass spectrometer 

(LC/MS) methods becomes a necessity for 

today's pharmaceutical laboratory. An analyst 

should question why a method with only three


reduce future problems are 

possible. Using the right 

columns is the key to easier 

methods development. 

Applications Notes 

components needs a 40-minute runtime. The 

analyst's productivity becomes hampered by 

an inefficient method and laboratory overhead 

increases due to wasted mobile phase, more 

waste disposal, and additional labor to meet 

the deadline. In response to the needs of 

today's pharmaceutical laboratories, Restek 

has introduced HPLC and LC/MS methods 

development and validation kits containing a 

variety of columns to resolve current chromatography problems. 

Commonly Asked Questions 

• What products are included in the kits? 

Each methods development kit package contains four columns (Allure C18, Allure 

Acidix, Allure Basix, and Ultra IBD) to provide a wide range of selectivity. The kit for 

HPLC methods development contains the most popular size for HPLC analysis 

(150x4.6mm). The LC/MS kit contains the columns most frequently used for LC/MS 

applications (50x2.1mm). 

Methods validation kits contain three columns, each packed from different lots of the 

stationary phase. The validation kits are available for each of Restek's four most 

popular phases found in the methods development kit package and in two dimensions - 

- 150x4.6mm for HPLC methods and 50x2.1mm for LC/MS methods. 

• How does Restek ensure reproducibility? 

To ensure that Restek's phases are reproducible, a minimum of three lots of a 

stationary phase are verified for consistency before any new HPLC column is 

commercially released. Each lot of material must meet stringent chromatographic 

reproducibility requirements before release, and each column is individually tested 

before shipment. 

• How do the different phases in the kits affect selectivity? 

The Allure C18 phase, Ultra IBD phase, Allure Basix phase, and the Allure 

Acidix phase provide a wide range of selectivity based upon the pKa or polarity of the 

analyte. The Allure C18 phase is ideal for retention of neutral, strongly lipophilic 

analytes. The high 27% carbon load of the Allure C18 phase results in maximum 

retention of lipophylic analytes. 

The IBD phase has many of the separation characteristics of a C18 phase and will 

behave much like a C18 phase in reverse phase mode, although selectivity often will 

be enhanced. The IBD phase's special functionality also can create additional retention 

http://www.restekcorp.com/hplc/methodkits.htm (2 of 3) [12/7/01 8:59:52 PM]


for analytes containing polar groups, cations, or anions, which can be poorly retained 

by C18 phases. 

For even greater retention of acids and bases, the Allure Acidix and Allure Basix 

phases, respectively, are the columns of choice. The Allure Acidix phase is 

engineered to provide maximum retention of acids and anions. The Allure Acidix 

phase retains amino acids without derivitization and will retain other short to medium 

chain length organic acids. The Allure Basix phase provides maximum retention of 

bases and cations. These stationary phases display their own unique retention profiles 

and do not mimic the Allure C18 phase at any point in the separation. It is important 

to remember that neither the Allure Acidix nor the Allure Basix phase are ion 

exchange packings. They retain analytes by polar interaction. 

Methods Development and Validation Kits 

Methods Development Kits cat.# 

HPLC (four 150x4.6mm columns) 25350 

LC/MS (four 50x2.1mm columns) 25351 

HPLC Methods Validation Kits 

(three 150x4.6mm columns) cat.# 

Allure C18 

25352 

Ultra IBD 

25355 

Allure Basix 

25354 


25353 

LC/MS Methods Validation Kits 

(three 50x2.1mm columns) cat.# 

Allure C18 

Ultra IBD 

Allure Basix 


25356 

25359 

25358 

25357 

• Can I order a kit customized to fulfill my 

company's specific analytical needs? 

Custom kits containing columns in the 

dimensions of your choice are available 

upon request. Kits that contain other 

phases are also available. For more 

information on custom methods 

development or validation kits, contact 

Restek's technical service department at 

814-353-1300 or 800-356-1688, ext. 4. 

http://www.restekcorp.com/hplc/methodkits.htm (3 of 3) [12/7/01 8:59:52 PM]

Restek -- HPLC Applications (PDF) 


HPLC Applications (PDF): 


LC/MS Analysis of Ascorbic Acid on Allure Acidix 

Amino Acids on Allure Acidix 

Adenine Nucleotides on the Allure Acidix 

Amino Acids on Allure Acidix 

Acetaminophen on Allure Acidix 

Aspirin and Salicylic Acid on Allure Acidix 

Ibuprofen and Naproxen on Allure Acidix 

Organic Acids on Allure Acidix 

Allure Basix 

LC-MS Analysis of Tricyclic Antidepressants on Allure Basix 

Ethinyl Estradiol and Norgestrel on Allure Basix 

Ethynyl Estradiol and Norethindrone on Allure Basix 

Ibuprofen and Pseudoephedrine on Allure Basix 

Caffeine, Acetaminophen, and Pseudoephedrine on Allure Basix 

Timolol Maleate on Allure Basix 

Enalapril Maleate on Allure Basix 

Metoprolol on Allure Basix 

Albuterol on Allure Basix 

Cyclobenzaprine HCl on Allure Basix 

Triamterene and Hydrochlorothiazide on Allure Basix 

Norethindrone and Medroxyprogesterone Acetate on Allure Basix 

Analgesics on Allure Basix 

Sertraline HCL on Allure Basix


Alprazolam and Lorazepam on Allure Basix 

Verapamil HCL on Allure Basix 

Triamterene on Allure Basix 

Atenolol on Allure Basix 

Sulfamethoxazole on Allure Basix 

Trycyclic Antidepressants on Allure Basix 

Allure C18 

Explosives (non-8330) on Allure C18 

Norgestrel and Ethynyl Estradiol on Allure C18 



Trimethoprim and Sulfamethoxazole on Allure C18 

EPA Explosives Method 8330 on Allure C18 

Corticosteroids on Allure C18 

Allure PFP-propyl 

Benzaldehyde, Hydrocortisone, and Prednisolone on Allure PFP-propyl-n 

Cocaine (COC) and Ecgonine Methyl Ester (EME) on Allure PFP Propyl 

Ultra IBD 

Carbamtes on Ultra IBD 

Uracil, Uridine, UTP, UDP, and UMP on Ultra IBD 

Thymine, Thymidine, TTP, TDP, and TMP on Ultra IBD 

Guanine, Guanosine, GTP, GDP, and GMP on Ultra IBD 

Cytosine, Cytidine, CTP, CDP, and CMP on Ultra IBD 

Adenine, Adenosine, ATP, ADP, and AMP on Ultra IBD 

11 Component Mixture of Nucleotides, Nucleosides, and Nitrogenous Bases on Ultra IBD 

LC/MS of Penicillin V on Ultra IBD 

LC/MS of Cephradine on Ultra IBD 

LC/MS of Cephaloridine on Ultra IBD 

LC/MS Analysis of Cimetidine on Ultra IBD 

Cephaloridine by Reverse Phase on Ultra IBD 

Cephaloridine Normal Phase Separation on Ultra IBD 

Penicillin V on Ultra IBD


Cephalexin and Cephradine on Ultra IBD 

Amoxicillin on Ultra IBD 

Digoxin on Ultra IBD 

Diflubenzuron on Ultra IBD 

Furosemide on Ultra IBD 

Pyridine and Phenol on Ultra IBD at pH 7.0 

Acids and Bases at pH 3.0 on Ultra IBD 

Acids and Bases at pH 7.0 on Ultra IBD 

Ultra PFP 

Levothyroxine on Ultra PFP 

Ultra Phenyl 

Guaifenesin with Codeine on Ultra Phenyl 

Ultra Cyano 

Epa 8330 Explosives on Ultra Cyano 

Beclomethasone on Ultra Cyano 

Calcium Channel Blockers on Ultra Cyano 

Ultra C4 

Hormones on Ultra C-4 

Ultra Aqueous C18 

Organic Acids on Ultra Aqueous C18 

Ultra C18 

EPA Method 8316 on Ultra C18 

Carbonyls: DNPH derivatives on Ultra C18 

ß-Estradiol, Norethinedrone, and Ethynyl Estradiol on Ultra C18 

Beclomethasone and des-chlorobeclomethasone on Ultra C18 



ß-Estradiol, Norethinedrone, and Ethynyl Estradiol on Ultra C18 

Corticoids on Ultra C18 

EPA Method 8316 on Ultra C18 

Carbonyls: DNPH derivatives on Ultra C18 

Carbonyls on Ultra C18


Pinnacle Specialty Phases 

Carbonyls on Pinnacle TO-11 

Pinnacle EcoSep exhibits excellent selectivity for uron herbicides. 

The Pinnacle Carbamate column offers optimized selectivity for free carbamate 

pesticides. 

http://www.restekcorp.com/hplcapps/ (4 of 4) [12/7/01 9:01:48 PM]

Ascorbic Acid on 

Allure Acidix (LC/MS) 

Peak List: 

1. unknown (2.33 min) 

2. ascorbic acid (8.84 min) 

Sample: 

Inj.: 5µL 

Conc.: 50µg/mL 

Solvent: water:methanol (1:1, v/v) 

2 

Application Note: LC_0114 

Column: Allure Acidix 

Catalog #: 9162565 

Dimensions: 150 x 4.6mm 

Particle Size: 5µm 


Conditions: 

Mobile Phase: 

Flow: 

Temp.: 

Det.: 

Interface: 

Ion Mode: 

5 mM ammonium formate, 

pH 3 :methanol (50:50, v/v) 

1.0mL/min 

ambient 

PE/Sciex API 150 EX 

Heated Nebulizer 

negative 

Needle current: 4µA 

Temp.: 350°C 

Orifice: -30V 

Ring: -300V 

Auxiliary gas: 6000 cc/min 

1 

0 1 2 3 4 5 6 7 8 9 min. 

Restek Corporation 110 Benner Circle Bellefonte, PA 16823 814-353-1300 800-356-1688 Fax: 814-353-1309

Amino Acids on 



Peak List: 

1. phenylalanine 

2. methionine 

3. tyrosine 

Sample: 

Inj.: 5µL 

Conc.: 0.3 mg/mL 

Solvent: mobile phase 

1 

2 


Catalog #: 9162565 




Conditions: 

Mobile Phase: 20mM KH 2 

PO 4 

pH3: 

acetonitrile (25:75, v/v) 

Flow: 1.0mL/min 

Temp.: 30°C 

Det.: UV @ 210nm 

3 

0 2 4 6 8 min. 

Restek Corporation 110 Benner Circle Bellefonte, PA 16823 • 814-353-1300 • 800-356-1688 • Fax: 814-353-1309 • www.restekcorp.com

Adenine Nucleotides 

on Allure Acidix 

Peak List: 

1. adenosine 

2. adenine 

3. adenosine 5'-monophosphate 

4. adenosine 5'-diphosphate 

5. adenosine 5'-triphosphate 

Sample: 

Inj.: 5µL 



1 

2 

3 



Catalog #: 9162565 




Conditions: 

Mobile Phase: 20mM KxHyPO4, 

pH 7:methanol ( 85:15, v/v ) 


Temp.: 35°C 


4 

5 

0 1 2 3 4 5 6 7 8 9 min. 


Amino Acids on 

Application Note:0064 


Peak List: 

1. Phenylalanine 

2. Methionine 

3. Tyrosine 

Inj. Vol.: 5µl 

Conc.: (0.3 mg/mL each) 

Solvent: Mobile Phase 

1 

2 


Catalog #: 9162565 

Dimensions: 150 x 4.6 mm 

Particle Size: 5 µm 


Conditions: 

Mobile Phase: 20mM KH2PO4 pH3: 

Acetonitrile: 

(25:75 v/v) 

Flow: 1.0ml/min 

Temperature: 30C 

Detection: UV @ 210nm 

3 

0 2 4 6 8 min. 


Acetaminophen 


Peak List: 

1. acetaminophen 

Sample: 

Inj.: 10µL 

Conc.: 1mg/mL 

Solvent: water:acetonitrile 

(1:1, v/v) 

1 



Catalog #: 9162565 




Conditions: 

Mobile Phase: 20mM ammonium 

acetate pH 4.5: 

acetonitrile 

(95:5, v/v) 

Flow: 1.0mL/min. 

Temp.: ambient 


0 1 2 3 4 5 6 7 min. 


Aspirin and 


Salicylic Acid 


3 


Catalog #: 9162565 




Peak List: 

1. toluene 

2. unknown 

3. salicylic acid 

4. aspirin 

Sample: 

Inj.: 10µL 


Solvent: buffer : acetonitrile 

(1:1, v/v) 

1 

4 

Conditions: 



acetonitrile 

(20:80, v/v) 




2 

0 2 4 6 8 10 min. 


Ibuprofen and 

Naproxin on 


4 



Catalog #: 9162565 




Peak List: 

1. unknown 

2. unknown 

3. uracil 

4. ibuprofen 

5. naproxen 

Sample: 

Inj.: 10µL 

Conc.: 100µg/mL (ibuprofen 

and naproxen) 

Solvent: ammonium acetate 

buffer : acetonitrile 

(1:1, v/v) 

5 

Conditions: 



acetonitrile 

(30:70, v/v) 




1 2 

3 

0 1 2 3 4 5 6 7 min. 


Organic Acids 


2 



Catalog #: 9162565 




Peak List: 

1. pyridine 

2. benzoic acid 

3. ascorbic acid 

Sample: 

Inj.: 25µL 

Conc.(acids): 250µg/mL 


(99:1, v/v) 

1 

Conditions: 

Mobile Phase: 10mM potassium 

phosphate pH 2.6 : 



Temp.: 27.0°C 


3 

0 2 4 min. 


Tricyclic Antidepressants 

on Allure Basix (LC/MS) 

Peak List: 

1. desipramine 

2. trimipramine 

Sample: 

Inj.: 10µL 


Solvent: water:methanol (1:1, v/v) 

1 

2 


Column: Allure Basix 

Catalog #: 9161565 




Conditions: 

Mobile Phase: 

5mM ammonium formate, 

pH 3:methanol (25:75, v/v) 

Flow: 1.0mL/min, 

Split ratio: 1:1 

Column Temp.: ambient 

Det.: 


Interface: Heated Nebulizer 

Ion Mode: negative 

Ion Spray Voltage: 3000V 

Temp.: 400°C 

Orifice: -40V 

Ring: -100V 

Auxiliary gas: 6000cc/min 

0 1 2 3 4 5 6 7 8 9 min. 


Ethynyl Estradiol 

and Norgestrel 

on Allure Basix 

Peak List: 

1. uracil (marker) 

2. norgestrel 

3. ethynyl estradiol 

Sample: 

Inj.: 5µL 



2 

3 



Catalog #: 9161565 




Conditions: 

Mobile Phase: water: 

methanol 

(40:60, v/v) 

Flow: 1mL/min. 

Temp.: 25°C 


1 

0 1 2 3 4 5 6 7 8 9 min. 


Ibuprofen and 

1 


pseudoephedrine 



Catalog #: 9161565 




Peak List: 


2. ibuprofen 

3. pseudoephedrine 

Sample: 

Inj.: 5µL 



2 

3 

Conditions: 


acetate, pH 6.9: 

methanol 

(75:25, v/v) 


Temp.: 25°C 


0 2 4 6 8 10 12 14 min. 


Acetaminophen, 

Caffeine and 

psuedoephedrine 


Peak List: 

1. sample solvent (marker) 

2. caffeine 

3. acetaminophen 

4. psuedoephedrine 

Sample: 

Inj.: 5µL 



2 

3 



Catalog #: 9161565 




Conditions: 



methanol 

(75:25, v/v) 


Temp.: 25°C 


4 

1 

0 2 4 6 8 10 12 min. 


Timolol Maleate 


Peak List: 

1. maleate salt 

2. timolol 

Sample: 

Inj.: 

Conc.: 

Solvent: 

2.5µL 

1.2mg/mL 

methanol:water 

(30:70) 

2 



Catalog #: 9161565 




Conditions: 


formate, pH 2.5: 

acetonitrile 

(20:80, v/v) 


Temp.: 25°C 


1 

0 2 4 6 8 10 12 14 min. 


Enalapril Maleate 


Peak List: 

1. maleate salt 

2. enalapril 

Sample: 

Inj.: 

Conc.: 

Solvent: 

2.5µL 

1.1mg/mL 


(30:70) 

1 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(20:80, v/v) 


Temp.: 25°C 


2 

0 2 4 6 8 10 12 14 min. 


Metoprolol 


Peak List: 

1. unknown 

2. metoprolol 

Sample: 

Inj.: 

Conc.: 

Solvent: 

2.5µL 

1.5mg/mL 


(30:70) 

1 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(10:90, v/v) 


Temp.: 25°C 


0 2 4 6 8 10 12 14 min. 


Albuterol 


1 



Catalog #: 9161565 




Peak List: 

1. albuterol 

Sample: 

Inj.: 

Conc.: 

Solvent: 

2.5µL 

1.1mg/mL 


(30:70) 

Conditions: 



acetonitrile 

(20:80, v/v) 


Temp.: 25°C 


0 1 2 3 4 5 min. 


Cyclobenzaprine 


Peak List: 

1. toluene (marker) 

2. cyclobenzaprine HCl 

Sample: 

Inj.: 1µL 

Conc.: 1.5mg/mL 

Solvent: 100% water 

2 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(25:75, v/v) 


Temp.: 30°C 


1 

0 1 2 3 4 5 6 min. 


Triamterene and 

Hydrochlorothiazide 


Peak List: 

1. hydrochlorothiazide 

2. triamterene 

Sample: 

Inj.: 1µL 


Solvent: methanol:water: 

tetrahydrofuran 

(30:35:35) 

1 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(65:35, v/v) 


Temp.: 25°C 


2 

2.5 2.75 3 3.25 3.5 3.75 4 4.25 4.5 min. 


Norethindrone and 

1 


Medroxprogesterone 

Acetate on 

Allure Basix 

Peak List: 


2. norethindrone 

3. medroxyprogesterone acetate 

2 

3 


Catalog #: 9161565 




Conditions: 


acetonitrile 

(40:60, v/v) 


Temp.: 30°C 


Sample: 

Inj.: 10µL 



0 0.5 1 1.5 2 2.5 3 3.5 4 4.5 min. 


Analgesics 


Peak List: 

1. acetaminophen (3.7) 

2. codeine HCl(5.4) 

3. oxycodone HCl(6.3) 

4. hydrocodone bitartrate (7.6) 

Sample: 

Inj.: 5µL 



1 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(85:15, v/v) 


Temp.: 30°C 


2 

3 

4 

0 1 2 3 4 5 6 7 8 9 min. 


Sertraline HCI 


Peak List: 


2. sertraline HCI 

Sample: 

Inj.: 1µL 

Conc.: 1mg/mL 


(7:3, v/v) 

2 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(40:60, v/v) 


Temp.: 25.0°C 


1 

0 2 4 6 8 10 12 14 min. 


Alprazolam and 

Lorazepam 


Peak List: 


2. lorazepam 

3. alprazolam 

Sample: 

Inj.: 10µL 



(7:3, v/v) 

2 

3 



Catalog #: 9161565 




Conditions: 


formate pH 3.0: 

acetonitrile 

(30:70, v/v) 


Temp.: 25.0°C 


1 

0 1 2 3 4 5 min. 


Verapamil HCL 


Peak List: 


2. verapamil HCL 

Sample: 

Inj.: 1µL 

Conc.: 1mg/mL 


(7:3, v/v) 

2 



Catalog #: 9161565 




Conditions: 



acetonitrile 

(65:35, v/v) 


Temp.: 25.0°C 


1 

0 1 2 3 4 5 6 7 8 9 min. 


Triamterene on 

Allure Basix 

Peak List: 


2. triamterene 

Sample: 

Inj.: 1µL 

Conc.: 1mg/mL 

Solvent: methanol: 

tetrahydrofuran 

(7:3, v/v) 

2 



Catalog #: 9161565 




Conditions: 





Temp.: 25.0°C 


1 

0 2 4 6 8 10 min. 


Atenolol on 

Allure Basix 

Peak List: 


2. atenolol 

Sample: 

Inj.: 1µL 

Conc.: 1mg/mL 

Solvent: water:methanol 

(7:3, v/v) 

2 



Catalog #: 9161565 




Conditions: 





Temp.: 25.0°C 


1 

0 2 4 6 8 10 12 min. 


Sulfamethoxazole 


Peak List: 

1. sulfamethoxazole 

Sample: 

Inj.: 10µL 



(1:1, v/v) 

1 



Catalog #: 9161565 




Conditions: 

Mobile Phase: water:methanol 

(70:30, v/v) 




0 2.5 5 7.5 10 12.5 15 17.5 20 min. 


Trycyclic Antidepressants 


Peak List: 

1. doxepin 

2. amitriptyline 

3. imipramine 

4. nordoxepin 

5. nortriptyline 

6. desimipramine 

7. protriptyline 

Inj. Vol.: 10µL 

Conc.: (100 µg/mL each) 


1 

2 

3 

4 


Catalog #: 9161565 




Conditions: 

Mobile Phase: 5mM KH2PO4 pH3: 

Methanol:Acetonitrile 

(15:25:60 v/v) 


Temperature: 30°C 


6 

5 

7 

0 1 2 3 4 5 6 7 8 9 min. 


Explosives on 

Allure C18 

Peak List: 

1. HMX 

2. RDX 

3. 1,3,5-trinitrobenzene 

4. 1,3-dinitrobenzene 

5. tetryl 

6. nitrobenzene 

7. 4-amino-2,6-dinitrotoluene 

8. 2-amino-4,6-dinitrotoluene 

9. trinitrotoluene 

10. 2,6-dinitrotoluene 


12. 2-nitrotoluene 



1 

2 

3 

4 

5 

6 

9 

8 

7 

10 

11 


Column: Allure C18 

Catalog #: Custom Number 55582 




Conditions: 

Mobile Phase: water: methanol 

(45:55, v/v) 


Temp.: 45°C 


Sample: 

Inj.: 5µL 


Solvent: water:methanol (1:1) 

12 

13 

14 

0 2 4 6 8 10 min. 


Norgestrel and 


Ethynyl Estradiol 

on the Allure C18 

Peak List: 

1. uracil (void marker) 


3. norgestrel 

Sample: 

Inj.: 5µL 


Solvent: methanol:water (60:40) 

1 

2 


Catalog #: 9164565 




Conditions: 

Mobile Phase: water:methanol (25:75) 

Flow: 

1.0 mL/min 

Temp.: 25°C 

Det.: 

UV @ 220nm 

3 

0 1 2 3 4 5 min. 


Norgestrel and Ethynyl 

Estradiol on the Allure C18 

Peak List: 



3. norgestrel 

Sample: 

Inj.: 5µL 


Solvent: methanol:water 

(60:40) 

1 

2 



Catalog #: 9164565 




Conditions: 


Flow: 

1.0 mL/min 

Temp.: 25°C 

Det.: 

UV @ 220nm 

3 

0 5 10 15 20 25 min. 


Norgestrel and Ethynyl 

Estradiol on the Allure C18 

Peak List: 



3. norgestrel 

Sample: 

Inj.: 5µL 



(60:40) 

1 

2 



Catalog #: 9164565 




Conditions: 


Flow: 

1.0 mL/min 

Temp.: 25°C 

Det.: 

UV @ 220nm 

3 

0 2.5 5 7.5 10 12.5 15 17.5 20 min. 


Trimethoprim and 

Sulfamethoxazole 

on Allure C18 

Peak List: 

1. trimethoprim 

2. sulfamethoxazole 

3. naphthoate 

Sample: 

Inj.: 10µL 



(1:1, v/v) 

1 

2 



Catalog #: 9164565 




Conditions: 

Mobile Phase: 10mm ammonium 

formate, pH to 3.0 

with formic acid: 

methanol (50:50, v/v) 




3 

0 2 4 6 8 10 12 14 min. 


EPA Method 8330 


Peak List: 

1. HMX 

2. RDX 

3. 1,3,5-TNB 

4. 1,3-DNB 

5. NB 

6. Tetryl 

7. 3,4-DNT (Int. std.) 

8. 2,4,6-TNT 

9. 4-Am-DNT 

10. 2-Am-DNT 

11. 2,6-DNT 

12. 2,4-DNT 

13. 2-NT 

14. 4-NT 

15. 3-NT 


Conc.: (100 ug/ml each) 


1 

2 

3 

4 

5 

6 

7 

8 

9 

10 

11 

12 


Catalog #: 9164575 




Conditions: 

Mobile Phase: Water:Methanol 

(50:50 v/v) 

Flow: 1ml/min 

Temperature: ambient 


13 

14 

15 

0 5 10 15 20 25 30 35 40 

minutes 


Corticosteroids 


Peak List: 

1. Uracil 

2. Corticosterone 

3. Deoxycorticosterone 


Conc.: (100 ug/ml each) 

Solvent 

1 


Catalog #: 9164565 




Conditions: 

Mobile Phase: Water:Methanol 

(50:50 v/v) 


Temperture: 


2 

3 

0 1 2 3 4 min. 


Benzaldehyde, Hydrocortisone, 

and Prednisolone on 1 

PFP-propyl-n 

(non-endcapped) 

Peak List: 

1. benzaldehyde 

2. hydrocortisone 

3. prednisolone 

Sample: 

Inj.: 5µL 




Column: Allure PFP Propyl-n 

(non-endcapped) 

Catalog #: custom 




Conditions: 

Mobile Phase: water:methanol (45:55, v/v) 


Temp.: 30°C 


2 3 

0 2 4 6 8 10 12 14 min. 


Cocaine (COC) and 

Ecgonine Methyl Ester (EME) 

on Allure PFP Propyl 

Peak List: 

1. EME (ecgonine methyl ester) 

2. COC (Cocaine) 

Sample: 

Inj.: 10µL 


Solvent: water 

Temp.: 4°C 

XIC of +MRM (3 pairs) for 200.0 / 182.0 amu 

XIC of +MRM (3 pairs) for 304.0 / 182.0 amu 

1 

0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min. 

0 0.5 1.0 1.5 2.0 2.5 3.0 3.5 4.0 4.5 min. 

2 

1.30e3 cps 

2.35e3 cps 


Column: 


Catalog#: 9169532 

Dimensions: 30mm x 2.1mm 

Particle size: 5µm 

Pore size: 60Å 

Conditions: 

Mobile phase: 

Flow: 

Column temp.: 

5mM pH 3.0 ammonium 

formate:acetonitrile (10:90) 

0.6mL/min. 

ambient 

Det.: PE/Sciex API 3000 

Interface: 

Turbo Ion Spray, ESI 

Interface temp.: 150°C 

Ion mode: 

positive 

ESI probe voltage: 5000V 

Orifice: 

+ 71V 

Ring: 

+ 265V 

Collision gas: 

Nitrogen 

Collision gas pressure: 2.2 mTorr 

Collision gas energy(COC): 28 eV 

Collision gas energy(EME): 26 eV 

Electron multiplier: 2100 volts 

Auxillary gas flow: 7000cc/min. 

Nebulizer gas setting: 15lb/in. 2 

Curtain gas setting: 12lb/in. 2 

Data Courtesy of Shane Needham, Pfizer Inc 


Carbamates on Ultra IBD 

Peak List: 

1. Aldicarb Sulfoxide 

2. Aldicarb Sulfone 

3. Oxamyl 

4. Methomyl 

5. 3-hydroxycarbofuran 

6. Aldicarb 

7. Propoxur 

8. Carbofuran 

9. Carbaryl 

10. Methiocarb 

Sample: 

Inj.: 5µL 

Conc.: 1 mg/mL 

each component 

Solvent: Methanol 

9 


Column: Ultra IBD 

Catalog #: 9175565 




Conditions: 

Mobile Phase: 

Flow: 

Temp.: 

Det.: 

Gradient 

A: Water 

B: Methanol 

Initial: 80% A: 20% B 

15 min.: 20% B: 80% B 

1.0mL/min 

ambient 

UV @ 220nm 

10 

1 2 

3 

4 

5 

6 

7 8 

0 2 4 6 8 10 12 14 16 18 min. 


Uracil, Uridine, UTP, UDP, 

and UMP on Ultra IBD 

Peak List: 

1. UTP 

2. UDP 

3. UMP 

4. Uracil 

5. Uridine 

Sample: 

Inj.: 20µL 

Conc.: 90-360 µg/mL 


Solvent: 20mM Ammonium 

Acetate, pH 5.8 

Component: Conc. (µg/mL): 

1. UTP 360 

2. UDP 360 

3. UMP 284 

4. Uracil 90 

5. Uridine 230 

2 

1 

3 

4 

5 



Catalog #: 9175565 




Conditions: 

Mobile Phase: 

97.5:2.5 20mM 

Ammonium Acetate, 

pH 5.8:Methanol 

Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 

0 2 4 6 8 10 12 14 16 18 min. 


Thymine, Thymidine, TTP, 

TDP, and TMP on Ultra IBD 

Peak List: 

1. TTP 

2. TDP 

3. TMP 

4. Thymine 

5. Thymidine 

Sample: 

Inj.: 20µL 

Conc.: 88-346µg/mL 




2 

3 

4 

5 



Catalog #: 9175565 




Conditions: 

Mobile Phase: 

97.5:2.5 20mM 



Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 


1. TTP 224 

2. TDP 152 

3. TMP 346 

4. Thymine 88 

5. Thymidine 318 

1 

0 2 4 6 8 10 12 14 16 18 min. 


Guanine, Guanosine, GTP, 

GDP, and GMP on Ultra IBD 

Peak List: 

1. GTP 

2. GDP 

3. GMP 

4. Guanine 

5. Guanosine 

Sample: 

Inj.: 20µL 

Conc.: 80-362µg/mL 




2 

3 



Catalog #: 9175565 




Conditions: 

Mobile Phase: 

97.5:2.5 20mM 



Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 


1. GTP 212 

2. GDP 260 

3. GMP 362 

4. Guanine 80 

5. Guanosine 226 

1 

4 

5 

0 2.5 5 7.5 10 12.5 15 17.5 min. 


Cytosine, Cytidine, CTP, 

CDP, and CMP on Ultra IBD 

Peak List: 

1. CTP 

2. CDP 

3. CMP 

4. Cytosine 

5. Cytidine 

Sample: 

Inj.: 20µL 

Conc.: 100-340 µg/mL 




2 

4 

5 



Catalog #: 9175565 




Conditions: 

Mobile Phase: 

97.5:2.5 20mM 



Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 


1. CTP 248 

2. CDP 322 

3. CMP 248 

4. Cytosine 100 

5. Cytidine 340 

1 

3 

0 2 4 6 8 10 12 14 16 18 min. 


Adenine, Adenosine, ATP, 

ADP, and AMP on Ultra IBD 

Peak List: 

1. ATP 

2. ADP 

3. AMP 

4. Adenine 

5. Adenosine 

Sample: 

Inj.: 20µL 

Conc.: 84-320 µg/mL 




1 

2 

3 

5 



Catalog #: 9175565 




Conditions: 

Mobile Phase: 

97.5:2.5 20mM 



Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 


1. ATP 258 

2. ADP 320 

3. AMP 274 

4. Adenine 84 

5. Adenosine 254 

4 

0 2 4 6 8 10 12 14 16 18 min. 


11 Component Mixture of 

Nucleotides, Nucleosides, 

and Nitrogenous Bases 

on Ultra IBD 

5 



Catalog #: 9175565 




Peak List: 

1. CDP 

2. CMP 

3. ATP 

4. ADP 

5. Cytidine 

6. TMP 

7. AMP 

8. Guanine 

9. Guanosine 

10. Adenine 

11. Adenosine 

Sample: 

Inj.: 20µL 

Conc.: 38-157µg/mL each component 

Solvent: 20mM Ammonium Acetate, pH 5.8 

2 

1 

3 

4 

6 7 

8 

9 

10 

Conditions: 

Mobile Phase: 

97.5:2.5 20mM 



Flow: 

1.0mL/min 

Temp.: 35°C 

Det.: 

UV @ 260nm 

11 


1. CDP 146 

2. CMP 113 

3. ATP 117 

4. ADP 145 

5. Cytidine 155 

6. TMP 157 


7. AMP 125 

8. Guanine 36 

9. Guanosine 103 

10. Adenine 38 

11. Adenosine 115 

0 2 4 6 8 10 12 14 16 18 min. 


LC/MS of 

Penicillin V 

on Ultra IBD 



Catalog #: 9175552 




Peak List: 

1. Penicillin V 

Sample: 

Inj.: 5µL 



1 

Conditions: 

Mobile Phase: 

5mM Ammonium 

Acetate pH 7.4: 

Acetonitrile(20:80) 

0.2mL/min 

ambient 


Turbo Ion Spray 

Positive 

8 L/hour 

12 L/Hour 

Flow: 

Temp.: 

Det.: 

Interface: 

Ion Mode: 

Nebulizer: 

Curtain Gas: 

Ion Spray Voltage: 4700.0v 

Temp.: 350 °C 

Orifice: 

+ 10.0 V 

Ring: 

+ 25.0 V 

SIM monitoring: 

349 ± 3dal. 

0 1.0 2.0 3.0 min. 


LC/MS of 

Cephradine 

on Ultra IBD 



Catalog #: 9175552 




Peak List: 

1. Cephradine 

Sample: 

Inj.: 5µL 



1 

Conditions: 

Mobile Phase: 

5mM Ammonium 



0.2mL/min 

ambient 



Positive 

8 L/hour 

12 L/Hour 

Flow: 

Temp.: 

Det.: 

Interface: 

Ion Mode: 

Nebulizer: 

Curtain Gas: 


Temp.: 350 °C 

Orifice: 

+ 10.0 V 

Ring: 

+ 25.0 V 


349 ± 3dal. 

0 1.0 2.0 3.0 min. 


LC/MS of 

Cephaloridine 

on Ultra IBD 

Peak List: 

1. Cephaloridine 

Sample: 

Inj.: 5µL 



1 



Catalog #: 9175552 




Conditions: 

Mobile Phase: 

5mM Ammonium 



0.2mL/min 

ambient 



Positive 

8 L/hour 

12 L/Hour 

Flow: 

Temp.: 

Det.: 

Interface: 

Ion Mode: 

Nebulizer: 

Curtain Gas: 


Temp.: 350 °C 

Orifice: 

+ 10.0 V 

Ring: 

+ 25.0 V 


416 ± 3dal. 

0 1.0 2.0 3.0 min. 


Cephaloridine by 

Normal Phase on 

Ultra IBD 

Peak List: 

1. cephaloridine 

Sample: 

Inj.: 5µL 

Conc: 1mg/mL 

Solvent: acetonitrile:water (50:50) 

1 



Catalog #: 9175565 




Conditions: 

Mobile Phase: acetonitrile: pH 4.0 20mM 

ammonium phosphate (80:20,v/v) 


Temp.: 27°C 


0 1 2 3 4 5 6 7 8 9 min. 


Penicillin V 

on Ultra IBD 

Peak List: 

1. unknown 

2. unknown 

3. unknown 

4. penicillin V 

Sample: 

Inj.: 

Conc.: 

Solvent: 

2.5µL 

1.2mg/mL 

acetonitrile:water 

(10:90, v/v) 

4 



Catalog #: 9175565 




Conditions: 



acetonitrile 

(60:40, v/v) 


Temp.: 30°C 


1 

2 3 

0 1 2 3 4 5 6 7 8 min. 


Cephalexin 

and Cephradine 

on Ultra IBD 

Peak List: 

1. cephalexin 

2. cephradine 

Sample: 

Inj.: 10µL 


Solvent: acetonitrile:water 

(10:90, v/v) 

1 

2 



Catalog #: 9175565 




Conditions: 



acetonitrile 

(90:10, v/v) 


Temp.: 30°C 


0 2 4 6 8 10 min. 


Amoxicillin 

on Ultra IBD 

Peak List: 

1. amoxicillin 

Sample: 

Inj.: 5µL 



(10:90, v/v) 

1 



Catalog #: 9175565 




Conditions: 



acetonitrile 

(95:5, v/v) 


Temp.: 30°C 


0 1 2 3 4 5 6 min. 


Digoxin 

on Ultra IBD 

Peak List: 

1. unknown 

2. unknown 

3. digoxin 

Sample: 

Inj.: 10µL 



(1:1, v/v) 

3 



Catalog #: 9175565 




Conditions: 

Mobile Phase: water with 

0.1% (v/v) acetic acid: 



Temp.: 27°C 


1 

2 

0 1 2 3 4 5 6 7 8 9 min. 


Diflubenzuron 

on Ultra IBD 

Peak List: 

1. diflubenzuron 

2. N,N' bis-(chlorobenzyl) urea 

Sample: 

Inj.: 10µL 

Conc.: 1mg/mL 


(1:1, v/v) 

1 



Catalog #: 9175365 




Conditions: 


acetonitrile 

(45:55, v/v) 




2 

0 2 4 6 8 10 12 14 min. 


Furosemide 

on Ultra IBD 

Peak List: 


2. furosemide 

Sample: 

Inj.: 5µL 



(60:40, v/v) 

2 



Catalog #: 9175565 




Conditions: 





Temp.: 27.0°C 


Impurities in Furosemide 

(enlargement of baseline) 

1 2 

( ) 

1. uracil 

2. furosemide 

1 

0 2 4 6 8 10 12 14 min. 

0 2 4 6 8 10 12 14 min. 


Pyridine/Phenol @ pH7 


Peak List: 

1. Uracil 5µg/mL 

2. Pyridine 50µg/mL 

3. Phenol 300µg/mL 



1 

2 


Catalog #: 9175565 


Particle Size: 5µg/mg 5 µm 


Conditions: 

Mobile Phase:20mM KxHyPO4 pH7: 

Acetonitrile (80:20 v/v) 

Flow: 1ml/min 



3 

0 2 4 6 8 10 

minutes 


Acids and Bases @ pH3 


Peak List: 


2. Unknown 

3. Maleate 5µg/mL 

4. Benzoic Acid 50µg/mL 

5. Nortriptyline 50µg/mL 

6. Amitriptyline 50µg/mL 

7. Trimipramine 50µg/mL 



1 

4 

5 

6 

7 


Catalog #: 9175565 




Conditions: 

Mobile Phase:20mM KH2PO4 pH3: 


Flow: 1ml/min 



2 

3 

0 2 4 6 8 10 

minutes 


Acids and Bases @ pH7 


2 

3 

Peak List: 


2. Benzoic Acid 50µg/mL 

3. Maleate 5µg/mL 

4. Trimipramine 50µg/mL 

5. Nortriptyline 50µg/mL 

6. Amitriptyline 50µg/mL 


Solvent: 


Catalog #: 9175565 




Conditions: 

Mobile Phase:20mM KxHyPO4 pH7: 


Flow: 1ml/min 



1 

4 

5 

6 

0 5 10 15 20 25 

minutes 


Levothyroxine 

on Ultra PFP 

Peak List: 

1. unknown 

2. levothyroxine 

Sample: 

Inj.: 20µL 


Solvent: 

water:methanol 

(70:30) 

Add 3 drops of concentrated 

ammonium hydroxide to each 

10mL of the sample diluent. 

Levothyroxine will not dissolve 

unless the pH is basic. 

2 


Column: Ultra PFP 

Catalog #: 9199565 




Conditions: 



methanol 

(60:40, v/v) 


Temp.: 25°C 


1 

0 2 4 6 8 10 12 14 16 18 min. 


Guaifenesin 

1 

2 


and Codeine 

on Ultra Phenyl 

Column: Ultra Phenyl 

Catalog #: 9105565 




Peak List: 

1. codeine 

2. guaifenesin 

Sample: 

Inj.: 5µL 

Conc.: codeine 825µg/mL 

guaifenesin 375µg/mL 


(50:50) 

Conditions: 



acetonitrile 

(80:20, v/v) 


Temp.: 25°C 


0 1 2 3 4 5 6 min. 


EPA 8330 on 

Ultra Cyano 

Peak List: 

1. nitrobenzene 




5. 1,3,5-trinitrotoluene 

6. 1,3-dinitrobenzene 



9. 2,4,6-trinitrotoluene 

10. 4-amino, 2,6-dinitrotoluene 

11. 2-amino, 4,6-dinitrotoluene 

12. RDX 

13. tetryl 

14. HMX 

5 

1 

8 

6 

4 

3 

2 

7 9 

11 

10 

Enlargement 

8.208 

8.379 

8.797 

2, 3 4 5 

6 

9.086 

7 

8 9 10 min. 

9.877 


Column: Ultra Cyano 

Catalog #: 9106575 




Conditions: 


Flow: 

1.0mL/min 

Temp.: 

ambient 

Det.: 

UV @ 254nm 

Sample: 

Inj.: 5µL 



12 

13 

14 

0 10 20 30 40 50 60 min. 


Beclomethasone 

on Ultra Cyano 

Peak List: 

1. des-chlorobeclomethasone 

2. beclomethasone 

Sample: 

Inj.: 10µL 



(9:1, v/v) 

2 



Catalog #: 9106565 




Conditions: 

Mobile Phase: water:acetonitrile 

(70:30, v/v) 




1 

0 1 2 3 4 5 6 7 8 9 min. 


Calcium Channel Blockers 


Peak List: 

1. Diltiazem 

2. Nifedipine Impurity 

3. Verapamil 

4. Nifedipine 

5. Nicardipine 

Inj. Vol.: 5µL 


Solvent: 1:1 ACN:Water 

1 

4 

5 


Catalog #: 9106565 




Conditions: 

Mobile Phase: 20mM Potassium 

Phosphate Monobasic, 

pH 3.0:ACN (70:30v/v) 


Temperature: 30° C (± 1° C) 

Detection: 235 nm 

3 

2 

0 1 2 3 4 5 6 7 8 9 10 min. 


Hormones 

on Ultra C-4 

1 

2 


Column: Ultra C-4 

Catalog #: 9102365 




Peak List: 

1. ß-estradiol 

2. norethindrone 

3. 17α-ethinylestradiol 

Sample: 

Inj.: 15µL 



(70:30) 

3 

Conditions: 



acetonitrile 

(60:40, v/v) 


Temp.: 30°C 


0 2 4 6 8 10 12 14 min. 


Organic Acids on 


Ultra Aqueous C18 

Peak List: 

Sample Conc.: 

(mg/mL) 

1 

3 

4 

5 

Column: Ultra Aqueous C18 

Catalog #: 9178565 




1. Malonic Acid 500 

2. Lactic Acid 500 

3. Acetic Acid 1000 

4. Citric Acid 1000 

5. Succinic Acid 2000 

6. Fumaric Acid 10 

6 

Conditions: 

Mobile Phase: 

99:1 pH 2.5 50 mM 

Potassium Phosphate: 

Acetonitrile 

Flow: 

1.5mL/min 

Temp.: 25°C 

Det.: 

UV @ 210nm 

Sample: 

All analytes dissolved in 

HPLC grade water. 

2 

Inj.: 10µL 

0 1 2 3 4 5 min. 


EPA Method 8316 


1 

4 

Peak List: 

1. Acrylamide (50 ppm) 

2. Unknown 

3. Acrolein (100 ppm) 

4. Acrylonitrile (50 ppm) 

Inj. 5µl 

Conditions: 

Column: Pinnacle Ultra C18 

Catalog Number: 9174565 




3 

Solvent A: Water 60% 

Solvent B: Acetonitrile 40% 

Mode : Isocratic 

Flow Rate: 1.0 ml/min 

Wavelength: 195 nm 

2 

0.5 1.0 1.5 2.0 2.5 3.0 3.5 

minutes 


Carbonyls 


1 

2 

3 

4 

5 

6 

7 

Column: Ultra C18 

Catalog Number: 9174565 


Particle Size: 5 um 

Pore Size: 100A 

8 

9 

Peak List: 

DNPH Derivatives of 

1. Formaldehyde 

2. Acetaldehyde 

3. Acetone 

4. Acrolein 

5. Propionaldehyde 

6. Crotonaldehyde 

7. MEK 

8. Methacrolein 

9. Butyraldehyde 

10. Benzaldehyde 

11. Valeraldehyde 

12. m-tolualdehyde 

13. Hexanaldehyde 

Inj. 10ul (3.0 µg/ml as derivative) 

2 4 6 8 10 12 14 

minutes 

10 

11 

12 

13 

Conditions: 

Solvent A: Water 

Solvent B: Acetonitrile 

Solvent C: THF 

Mode : Gradient 

50 - 30% A: 0 - 15 min. 

30 - 65% B: 0 - 15 min. 

20 - 5% C: 0 - 15 min. 

Flow Rate: 1.5 ml/min 

Wavelength: 365 nm 

Temperature: 40 0 C 


β-Estradiol, Norethindrone 

and Ethynyl Estradiol 

on Ultra C18 

Peak List: 

1. β-estradiol 


3. norethinedrone 

Sample: 

Inj.: 10µL 


Solvent: water:acetonitrile (20:80) 

1 

3 



Catalog #: 9174565 




Conditions: 

Mobile Phase: 

pH 3.0 10mM 

ammonium formate: 

acetonitrile (40:60) 

Flow: 

1.0 mL/min 

Temp.: 30°C 

Det.: 

UV @ 230nm 

2 

0 1 2 3 4 min. 


Beclomethasone and 

des-chlorobeclomethasone 

on Ultra C18 

Peak List: 


2. des-chlorobeclomethasone 

Sample: 

Inj.: 10µL 

Conc.: 79.5 µg/mL (Beclomethasone) 


1 



Catalog #: 9174565 




Conditions: 

Mobile Phase: water:acetonitrile(30:70) 

Flow: 

1.0 mL/min 

Temp.: 25°C 

Det.: 

UV @ 238nm 

2 

0 0.5 1 1.5 2 min. 


Beclomethasone and 

des-chlorobeclomethasone 

on Ultra C18 

Peak List: 


2. deschlorobeclomethasone 

Sample: 

Inj.: 10µL 

Conc.: 79.5 µg/mL (Beclomethasone) 


1 



Catalog #: 9174565 




Conditions: 

Mobile Phase: water:acetonitrile(50:50) 

Flow: 

1.0 mL/min 

Temp.: 25°C 

Det.: 

UV @ 238nm 

2 

0 1 2 3 4 min. 


β-Estradiol, 

Norethindrone and 

Ethynyl Estradiol on 

Ultra C18 

2 



Catalog #: 9174565 




Peak List: 

1. β-estradiol 

2. norethinedrone 


Sample: 

Inj.: 10µL 


Solvent: acetonitrile:water (80:20) 

1 

3 

Conditions: 

Mobile Phase: 

pH 3.0 10mM 

ammonium formate: 

acetonitrile(60:40) 

Flow: 

1.0 mL/min 

Temp.: 30°C 

Det.: 

UV @ 230nm 

0 2 4 6 8 10 12 14 min. 


Corticoids on 

2 


Ultra C18 

Peak List: 

1. uracil 

2. triamcinolone 

3. hydrocortisone 

4. dexamethasone 

5. corticosterone 

6. deoxycorticosterone 

Sample: 

Inj.: 5µL 



3 

4 

5 

6 


Catalog #: 9174565 




Conditions: 

Mobile Phase: water:methanol 

(30:70, v/v) 

Flow: 

1.0mL/min 

Temp.: 30°C 

Det.: 

UV @ 220nm 

1 

0 1 2 3 4 5 6 min. 


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