Sunniest C18 Sunniest RP-AQUA Sunniest C8 Patent pending

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1 Patent pending Website : telno : in AUSTRALIA

2 A Novel Bonding Technique (patent pending) An unique trifunctional silyl-reagent was developed as shown in Fig.. This silyl-reagent can bond with any silanol groups on silica surface as shown in Fig. because it can expand and contract by itself. This technique can make residual silanol groups on C8 stationary phase to be the least amount. Finally an end-capping was done with trimethylsilyl-reagent reagent (TMS). X H C CH n X X n:,, or X: Cl, CH, CHCH etc. H H H TMS 化 Fig. Schematic diagram of bonding of novel silylreagent on silica surface Characteristics of Sunniest Features Fig. Novel C8 reagent Little residual silanol groups by an unique bonding technique Excellent stability, especially under acidic ph conditions Sharp peak shape for acidic, basic and chelating compounds RP-AQUA is available under 00% aqueous conditions, and shows enhanced retention of polar compounds. Particle size (μm) Pore diameter (nm) Specific surface area (m /g) Carbon content (%) Bonded phase ph range and 0 C and 0 Near C8-8 and 0 0 C Website : telno : in AUSTRALIA

3 Evaluation of End-capping Comparison of plates number (N) and USP tailing factor (TF) of amitriptyline CH H, ph7., 0 ºC N() =,000 TF() =.0 N() =0,00 TF() =.07 Column size: 0 x. mm Particle size: μm Mobile phase: CH H/0mM Phosphate buffer ph7. or.0 =80/0 CH CN/0mM Phosphate buffer ph7.0 =0/0 Flow rate:.0 ml/min Temperature: 0 ºC or ºC Sample: = Uracil = Propranolol CH NH CH Column A C8 N() =8,90 TF() =.7 = Nortriptyline H NH CH Column B C8 N() =0,000 TF() =.0 = Amitriptyline CH N CH CH CN, ph7.0, 0 ºC CH H, ph.0, ºC N() =9,000 TF() =.09 N() =,000 TF() =. N() =,800 TF() =.0 N() =8,000 TF() =.0 N() =,000 TF() =.09 Column A C8 N() =,900 TF() =.9 Column A C8 N() =,000 TF() =. Column B C8 N() =,800 TF() =.8 Column B C8 N() =,00 TF() = Amitriptyline is widely used to evaluate residual silanol groups on the C8 stationary phase. Peak shape of amitriptyline was compared under kinds of conditions such as methanol/phosphate buffer/0 ºC, methanol/phosphate buffer/ ºC and acetonitrile/phosphate buffer/0 ºC. Under methanol/phosphate buffer/0 ºC conditions which is the most common among HPLC manufacturers, good peak shape was obtained for all columns. There were little difference of a peak shape. Under acetonitrile/phosphate buffer/0 ºC, however, Sunniest columns showed a symmetrical peak, while column A and B C8 showed a terribly tailing peak., RP-AQUA and C8 columns allow to use a wide range of mobile phase without peak tailing because of extremely low content of residual silanol groups on the stationary phase. Website : telno : in AUSTRALIA

4 Stability under acidic and basic ph conditions Relative retention of ethylbenzene (%) B company C8 D company C8 F company C Time (h) Relative plate number of butylbenzene (%) , ph0 0ºC, ph9. 0ºC Time (h) Durable test condition Column size: 0 x. mm Mobile phase: CH CN/.0% TFA (ph) = 0/90 Flow rate:.0 ml/min Temperature: 80 ºC Measurement condition Column size: 0 x. mm Mobile phase: CH CN/H =0/0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Uracil = Ethylbenzene Durable test condition Column:, C8, μm 0 x. mm Mobile phase: C8: CH H/0mM Sodium borate/0mm NaH=0//9 (ph0) C8: CH H/0mM Sodium borate (ph9.) =0/70 Flow rate:.0 ml/min Temperature: C8-0 ºC, C8-0 ºC Measurement condition Column:, C8, μm 0 x. mm Mobile phase: CH CN/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Butylbenzene Stability under acidic ph conditions was evaluated at 80 ºC using acetonitrile/% trifluoroacetic acid solution (0:90) as mobile phase. 00% aqueous mobile phase expels from the pore of packing materials by capillarity and packing materials doesn t deteriorate. 0% acetonitrile in a mobile phase allows an accurate evaluation. -) has kept 90% retention for 00 hours under such severe conditions. ur bonding technique can make column life be long. is less stable than Sunniest C8. However, has more stable than other company C8 columns. ) N. Nagae, T. Enami and S. Doshi, LC/GC North America ctober 00. ) T. Enami and N. Nagae, American Laboratory ctober 00. ) T. Enami and N. Nagae, BUNSEKI KAGAKU, (00) 09. Stability under basic ph conditions was evaluated at 0 ºC using methanol/sodium borate buffer ph 0 (0:70) as mobile phase. Sodium borate is used as a alkaline standard solution for ph meter, so that its buffer capacity is high. Elevated temperature of 0 ºC makes column life be one third. When column is used at 0 ºC, column life becomes,000 hours. ther company shows stability test at ambient (room temperature). If room temperature is ºC, column life at room temperature ( ºC) is sixteen times longer than that at 0 ºC. is enough stable even if it is used under ph 0 condition. Regarding stability under basic ph condition, there is little C8 column like except for hybrid type C8. It is considered that our end-capping technique leads high stability. can be used at the ph range from. to 0. can be used at the ph range from. to 9. Website : telno : in AUSTRALIA

5 Relationship between ph and retention of acidic, basic and neutral compounds Basic compound (B) B N A ph Retention factor (k) Neutral compound (N) Acidic compound (A) A A N B N B ph7 ph0 ph ph selectivity, ph. ph7 ph0 Column:, μm 0 x. mm Mobile phase: A) 0mM Phosphoric acid ph. A) 0mM Phosphate buffer ph7 A) 0mM Phosphate buffer ph0 B) Acetonitrile Time (min) 0 0 %B (%) Flow rate:.0 ml/min Temperature: 0 ºC Detection: nm Sample: = Thiamine HCl Vitamin B = Nicotinamide = Nicotinic acid = Pyridoxine HCl Vitamin B = Folic acid = Riboflavin Vitamin B ph of mobile phase can make selectivity of ionic compounds change much. Sunniest C8 can be used at the ph range from. to 0, so that a suitable analytical method can be created using. Website : telno : in AUSTRALIA

6 Comparison data: Bleeding from column Comparison using Corona CAD Comparison using MS.E+08 F 社 C8 Area: 90,000.E+08.E+08.E+08 B 社 C8 B 社 C8 Area:,0,000 Intensity (cps).e+08.e+08.e+08.e+08 Area: 9,000.E+08.E+07 Column size: 0 x.0 mm Mobile phase: A) 0.% acetic acid B) CH CN Gradient: Time: 0min min.min 8min 9min %B: % % 00% 00% % Flow rate: 0. ml/min Temperature: 0 ºC Detection: Corona CAD 0.E Time (min) Column size: 0 x.0 mm Mobile phase: A) 0.% acetic acid B) CH CN Gradient: Time: 0min min.min 8min 9min %B: % % 00% 00% % Flow rate: 0. ml/min Temperature: 0 ºC MS: ABI API-000 Ionization: Turboionspray (cation) Measurement mode: Q Scan m/z Separation of antidepressants using acetonitrile and ammonium acetate a for LC/MS Company E C8 Company F C8 Company G C8 Company H C8 N=,00 TF=. N=,00 TF=.0 N=,000 TF=.8 N=8,00 TF=. N=,00 TF=. N=,00 TF=. N=8,00 TF=.0 Column size: 0 x. mm Particle size: μm Mobile phase: CH CN/0mM Ammonium acetate ph.8=0/0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: Amitriptyline CH N CH Website : telno : in AUSTRALIA

7 Stability Stability of under 00% aqueous conditions Relative retetion of ethylbenzene (%) Acidic ph test Time (h) Relative plate number of thtmine (%) Basic ph test Time (h) Durable test conditions Column:, μm 0 x. mm Mobile phase: 0.% TFA Flow rate:.0 ml/min Temperature: 0 ºC Durable test conditions Column:, μm 0 x. mm Mobile phase: 0mM Phosphate buffer ph8.0 Flow rate:.0 ml/min Temperature: 0 ºC Measurement conditions Column:, μm 0 x. mm Mobile phase: CH H/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Uracil = Amylbenzene Measurement conditions Column:, μm 0 x. mm Mobile phase: 0mM Phosphate buffer ph7.0 Flow rate:.0 ml/min Temperature: 0 ºC Sample: = Thymine It is important that stability is evaluated under 00% aqueous conditions for an AUQA type C8 column because column life becomes longer to increase a content of organic solvent in a mobile phase. can be used under 00% aqueous conditions from ph to ph8. can be used under 00% aqueous conditions from ph to ph 8. Sunniest RP- AQUA is one of the most stable aqua type column. Website : telno : in AUSTRALIA

8 Reproducibility of retention under 00% aqueous conditions C8 and C8 reversed phases exhibit decreased and poorly reproducible retention under more than 98% aqueous conditions as shown in Fig.. This problem traditionally has been explained as being the result of ligand collapse or a matting effect. Nagae - ascertained, however, that the mobile phase was being expelled from the pores of the packing material under 00% aqueous mobile phase conditions, as Fig. shows. When the surface of packing materials isn t wet by water, water used as a mobile phase expels from the pore of the packing material by capillarity. This is a reason why reproducibility in retention is low under 00% aqueous conditions. Reversely pressure around the packing material makes water permeate into the pore of the packing material to overcome a force worked by capillarity. In other words, the surface of a reversed phase like C8 isn t wet by water anytime even if water permeates into the pore of the packing material or not. So it is wrong that we say dewetting when water expel from the pore. Saying Depermetating is more suitable. is a reversed stationary phase, so that it is not wet with water. However the contact angle of water on the surface of is less than that of a conventional C8. Expelling force (pressure) acted by capillarity on is less than atmospheric pressure. So, shows reproducible retention under 00% aqueous conditions. After stopping pump for hour N. Nagae, T. Enami and S. Doshi, LC/GC North America ctober 00. Fig. Retention behavior of a C8 column under 00% aqueous mobile phase conditions lica Pressure 0nm Water Water permeates into the pore. Permeating Stop flow Water expels from the pore by capillarity. Depermenating Fig. Schematic diagram of C8 particle ) N. Nagae, T. Enami and S. Doshi, LC/GC North America ctober 00. ) T. Enami and N. Nagae, American Laboratory ctober 00. ) T. Enami and N. Nagae, BUNSEKI KAGAKU, (00) 09. Separation of nucleic acid bases 0 ºC ºC Column:, μm 0 x. mm Mobile phase: 0mM Phosphate buffer ph7.0 Flow rate:.0 ml/min Temperature: 0 ºC and ºC Sample: = Cytosine = Uracil = Thymidine = Uridine = Thymine Relative retention of thymine (%) Change of retention of thymine at 0 ºC (measurement every stop flow for hour) Measurement number ev ery stop flow showed more than 97% of reproducibility in retention using 00% aqueous buffer as a mobile phase. Website : telno : in AUSTRALIA

9 Separation of water-soluble vitamins Separation of nucleotides N=9, Column:, μm 0 x. mm Mobile phase: 0mM Phosphate buffer ph.8 Flow rate:.0 ml/min Temperature: 0 ºC Detection: Sample: = Nicotinic acid, = Pyridoxal = Pyridoxine, = Nicotinamide Column:, μm 0 x. mm Mobile phase: 0mM Phosphate buffer ph.0 Flow rate:.0 ml/min Temperature: 0 ºC Detection: Sample: = -GDP, = -GMP, = -ATP, = -ADP, = -AMP separation of olong tea Separation of water-soluble vitamins using mobile phase for LC/MS Caffeine xalic acid Theanine Guanylic acid Column:, μm 0 x. mm Mobile phase: A) H /H P (99.9:0.) B) CH CN Time (min) %B (%) Flow rate:.0 ml/min Temperature: 0 ºC Detection: nm Injection volume: 0 μl Sample: olong tea = (-)-Epigallocatechin = (+)-Catechin = (-)-Epigallocatechin gallate = (-)-Epicatechin = (-)-Epicatechin gallate Column:, μm 0 x. mm Mobile phase: A) 0mM Ammonium acetate B) Acetonitrile/ A solution (0:80) Time (min) 0 0 %B (%) Flow rate:.0 ml/min Temperature: 0 ºC Detection: nm Sample: = Nicotinic acid = Pyridoxine HCl Vitamin B = Nicotinamide = Thiamine HCl Vitamin B = Folic acid = Riboflavin Vitamin B Website : telno : in AUSTRALIA

10 Separation of standard samples Separation of pyridine and phenol α(pyridine/phenol) = 0.8 α(pyridine/phenol) = 0.0 Column:, μm 0 x. mm, μ m 0 x. mm, μm 0 x. mm Mobile phase: CH H/H =7/ Flow rate:.0 ml/min Temperature: 0 ºC Pressure:. MPa Sample: = Uracil, = Caffeine, = Phenol, = Butylbenzene, (N=,700) = o-terphenyl, (N=,000) = Amylbenzene, (N=,00) 7 = Triphenylene, (N=,00) N is plate number of the above. α(pyridine/phenol) = Column:, μm 0 x. mm, μ m 0 x. mm, μm 0 x. mm Mobile phase: CH H/H =0/70 Flow rate:.0 ml/min Temperature: 0 ºC Detection: Sample: = Uracil = Pyridine = Phenol C8 RP-AQUA C8 Hydrophobicity: α(amylbenzene/butylbenzene)... Hydrogen bonding capacity: α(caffeine/phenol) Steric selectivity: α(triphenylene/o-terphenyl).7.. Separation factor of pyridine and phenol is said to show the amount of residual silanol groups. The lower a value of separation factor, the less an effect of residual silanol groups. All Sunniest columns show one of the lowest value., RP-AQUA and C8 show not only high efficiency but also low column pressure. Website : telno : in AUSTRALIA

11 Separation of a chelating compound Separation of acidic compounds Column:, μm 0 x. mm, μ m 0 x. mm, μm 0 x. mm Mobile phase: CH CN/0mM H P =0/90 Flow rate:.0 ml/min Temperature: 0 ºC Detection: Sample: = 8-Quinolinol = Caffeine 0 8 Column:, μm 0 x. mm, μ m 0 x. mm, μm 0 x. mm Mobile phase: CH CN/0.% H P =/98 Flow rate:.0 ml/min Temperature: 0 ºC Detection: Sample: = Formic acid = Acetic acid = Propionic Acid, RP-AQUA, C8 are inert for a metal chelating compound and acidic and basic compounds, so that they show symmetrical peaks of each compound. Website : telno : in AUSTRALIA

12 Sunniest rdering information Inner diameter Length, μm, μm Sunniest RP- AQUA, μm Sunniest RP- AQUA, μm SunniestC8, μm, μm [mm] [mm] Catalog No. Catalog No. Catalog No. Catalog No. Catalog No. Catalog No..0 0 EB EB ER ER EC EC 7 EB EB - ER - EC - 00 EB ER ER EC EC 0 EB7 EB7 ER7 ER7 EC7 EC7.0 0 EB7 EB7 ER7 ER7 EC7 EC7 0 - EB8 - ER8 - EC8. 0 EB EB ER ER EC EC 0 EB EB ER ER EC EC 7 EB - ER - EC - 00 EB EB ER ER EC EC 0 EB7 EB7 ER7 ER7 EC7 EC7 0 - EB8 - ER8 - EC EB78 - ER78 - EC EB88 - ER88 - EC88 Manufacturer ChromaNik Technologies Inc. Website : telno : in AUSTRALIA / 0908

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