Amylopectin structure and crystallinity explains variation in digestion kinetics of starches across botanic sources in an in vitro pig model

Background: Starch is the main source of energy in commonly used pig diets.Besides effects related to the extent of starch digestion,also several effects related to variation in digestion rate have recently been demonstrated in non-ruminants.Different rates of starch digestion in animals and in in vitro models have been reported,depending on the botanic origin of starch.Starches from different botanic sources differ widely in structural and molecular properties.Predicting the effect of starch properties on in vitro digestion kinetics based on existing literature is hampered by incomplete characterization of the starches,or by a selective choice of starches from a limited number of botanic sources.This research aimed to analyse the relationships between starch properties and in vitro digestion kinetics of pure starches isolated from a broad range of botanic origins,which are used in non-ruminant diets or have a potential to be used in the future.Therefore we studied starch digestion kinetics of potato,pea,corn,rice,barley,and wheat starches,and analysed the granule diameter,number of pores,type and amount of crystalline structure,amylose content and amylopectin side-chain length of all starches.Results: Multivariate analysis revealed strong correlations among starch properties,leading us to conclude that effects of most starch characteristics are strongly interrelated.Across all analysed botanic sources,crystalline type and amylopectin chain length showed the strongest correlation with in vitro digestion kinetics.Increased percentages of A–type crystalline structure and amylopectin side chains of DP 6–24 both increased the rate of digestion.In addition,within,but not across,(clusters of) botanic sources,a decrease in amylose content and increase in number of pores correlated positively with digestion kinetics.Conclusion: The type of crystalline structure and amylopectin chain length distribution of starch correlate significantly with digestion kinetics of starches across botanic sources in an in vitro pig model.Variation in digestion kinetics across botanic sources is not additively explained by other starch properties measured,but appears to be confined within botanical sources.

Suppression of starch synthase I(SSI) by RNA interference alters starch biosynthesis and amylopectin chain distribution in rice plants subjected to high temperature

Based on known cDNAs of rice starch synthase isoforms,we constructed dsRNA interference vectors for starch synthase I(SSI)to produce transgenic plants containing starch with a moderately high amylose content.We investigated the effect of SSI suppression on grain quality traits,starch biosynthesis,and amylopectin chain distribution in rice plants exposed to two different temperature regimes.The activities and transcripts of BEs,DBEs,and other SS isoforms were further investigated to clarify the effect of SSI suppression on these key enzymes and their specific isoforms under different temperature treatments.Suppression of SSI by RNAi altered grain starch component and amylopectin chain distribution,but it exerted only a slight effect on total starch content(%)and accumulation amount(mg kernel?1)and on starch granule morphology and particle size distribution.Under normal temperature(NT),insignificant differences in kernel weight,chalky kernel proportion,chalky degree,and starch granule morphology between SSI-RNAi line and its wild type(WT)were observed.However,amylose content(AC)level and granule-bound starch synthase(GBSS)activity in rice endosperms were markedly increased by SSI-RNAi suppression.The chalky kernel proportion and chalky degree of SSIRNAi lines were significantly higher than those of WT under high temperature(HT)exposure at filling stage.Inhibition of SSI by RNAi affected amylopectin chain distribution and raised starch gelatinization temperature(GT)in two ways:directly from the SSI deficiency itself and indirectly by reducing BEIIb amounts in an SSI-deficient background.The deficiency of SSI expression led to an alteration in the susceptibility of grain chalkiness occurrence and starch gelatinization temperature to HT exposure,owing to a pleiotropic effect of SSI deficiency on the expression of other genes associated with starch biosynthesis.

Effects of dietary amylose to amylopectin ratio on growth performance,carcass quality characteristics and meat fatty acids in Chinese Qinchuan cattle

This study evaluated the effects of the dietary starch amylose/amylopectin ratio on growth performance,rumen fermentation and blood parameters,carcass characteristics and marbling score,and meat fatty acid profiles and transcriptional changes in the genes involved in the gluconeogenesis pathway in Chinese Qinchuan cattle.Forty-five cattle were randomly divided into three groups.The bulls were fed a control diet(middle amylose/amylopectin ratio=0.47)or diets with either waxy corn starch(WS)inclusion(low amylose/amylopectin ratio=0.23)or pea starch(PS)inclusion(high amylose/amylopectin ratio=0.60)for 90 days.The bulls were individually allowed to receive the diets at the daily amount of at least 2.1%of their individual BW twice daily.The bulls were weighed at 45-day intervals,and blood and rumen fluid samples were also collected at 45-day intervals.The bulls were slaughtered and longissimus thoracis(LT)samples were collected for meat quality measurements.Compared with PS,the average daily gain(ADG)and feed efficiency in control and WS were increased(P<0.05)during d 0 to 45.However,the feed efficiency in PS was increased(P<0.05)compared with the control and WS during d 46 to 90.During the whole feeding trial,the ADG in control and WS showed trends that were higher(0.05<P<0.1)than that of PS,while other growth performance measures and feed intake did not differ(P>0.05)among the three groups.Compared with PS,the serum insulin concentration in WS was increased(P<0.05)at d 45,but not at d 90.Compared with control and WS,the serum SOD and T-AOC concentrations in PS were increased(P<0.05)at d 45,but not at d 90.Compared with control,the rumen microbial crude protein contents in WS and PS were increased(P<0.05)at d 0,while those of control and WS were higher(P<0.05)than that of PS at d 45,but not at d 90.Compared with PS,the backfat thickness in control and WS were increased(P<0.05),however,the marbling score and expression of genes related to the gluconeogenesis pathway in liver of the three groups did not differ(P>0.05).Chemical and physio-chemical compositions of LT did not differ(P>0.05)among the three groups.In conclusion,diets with low or middle amylose/amylopectin ratios increased the backfat thickness,and tended to improve ADG,but had no effect on other carcass characteristics,fat content in the LT,or the expression of hepatic genes related to the gluconeogenesis pathway in Chinese Qinchuan cattle.

Amylopectin from Glutinous Rice as a Sustainable Binder for High-Performance Silicon Anodes

Silicon(Si)has been investigated as a promising anode material because of its high theoretical capacity(4200 m Ah g^(-1)).However,silicon anode suffers from huge volume changes during repeated charge–discharge cycles.In this work,inspired by a remarkable success of the glutinous rice mortar in the Great Wall with ca.2000-year history,amylopectin(AP),the key ingredient responsible for the strong bonding force,is extracted from glutinous rice and utilized as a flexible,aqueous,and resilient binder to address the most challenging drastic volume-expansion and pulverization issues of silicon anode.Additionally,the removal of toxic N-methyl-2-pyrrolidone(NMP)organic solvent makes the electrode fabrication process environmentally friendly and healthy.The as-prepared Si-AP electrode with 60 wt%of Si can uphold a high discharge capacity of 1517.9 m Ah g^(-1)at a rate of 0.1 C after 100 cycles.The cycling stability of the Si-AP has been remarkably improved in comparison with both traditional polyvinylidene fluoride(PVDF)and aqueous carboxymethylcellulose(CMC)binders.Moreover,when the content of silicon in the Si-AP electrode increases to 70 wt%,a high discharge capacity of 1463.1 m Ah g^(-1)can still be obtained after 50 cycles at 0.1°C.These preliminary results suggest that the sustainably available and environmentally benign amylopectin binders could be a promising choice for the construction of highly stable silicon anodes.

氮肥基追比对弱筋小麦宁麦9号籽粒淀粉合成及相关酶活性的影响

为研究弱筋小麦籽粒淀粉形成的机制,以弱筋小麦宁麦9号为材料,对不同氮肥基追比对其籽粒淀粉合成及相关酶活性的影响及相关生理基础研究表明,氮肥基追比7∶1∶2处理与5∶1∶4处理相比,花后7～28d籽粒直、支链淀粉及总淀粉积累速率低,成熟籽粒直链淀粉含量低,支链淀粉、总淀粉含量高,剑叶蔗糖磷酸合成酶(SPS)活性和蔗糖含量低,籽粒蔗糖合成酶(SS)活性低,籽粒蔗糖含量高,籽粒ADPG焦磷酸化酶(AGPP)、可溶性淀粉合成酶(SSS)、束缚态淀粉合成酶(GBSS)活性低,淀粉积累量低。相关分析表明,籽粒中AGPP、SSS、GBSS与直、支链及总淀粉积累速率、籽粒灌浆速率呈极显著正相关,说明这几种酶对促进籽粒淀粉的积累起重要作用。

Mechanism of Degradation of Rice Starch Amylopectin by Oryzenin Using ONIOM Quantum Calculations [DFT/B3LYP/6-31+G(D, P): AM1]

Understanding the molecular factors of rice degradation during its aging concerns our research team. This article emphasizes oryzenin-amylopectin. It aims to reveal the mechanism of amylopectin deterioration during rice aging. The research exploits the Natural Bond Analysis and ONION method at theory level DFT/B3LYP/6-31+G(d, p) and AM1. This methodological approach allows highlighting amylopectin transformation;oryzenin converts amylopectin into amyloidosis in continuous. This led to monosaccharides and disaccharides.

Hydrogen Bonds Sites of Amylose or Amylopectin from Starch at the ONION Level (B3LYP/6-311++G [d, p]: AM1)

The research aims to help reduce the enormous post-harvest losses of rice or bananas in Cote d’Ivoire. It focuses on the breakdown mechanism of their starch. This process contributes to their decay. This study wants to elucidate it;to do this, it analyzes the tetra or penta saccharide reactions with the water or dioxide carbon. It calculates the geometric, energetic and spectroscopic parameters at the ONIOM level (B3LYP/6-311++G [d, p]: AM1). These quantities allow establishing that oxygen <img src="Edit_7761c26d-f67d-4826-ab0e-8bff55d6faa3.png" alt="" /> represents the privileged hydrogen bond (HB) site for amylose with four or five D-glucose building blocks. They suggest <img src="Edit_ecd55400-653e-43d0-986d-9814259ebda7.png" alt="" /> and <img src="Edit_048b683f-5690-4078-b8ad-88fa25c5875c.png" alt="" /> are respectively those of amylopectin when the latter consists of four and five D-glucose synthons. They prove that amylose deteriorates before it;the degradation of starch begins with its alteration into disaccharides. It continues with that of amylopectin into tri or tetra-saccharides with four or five building blocks.

^(60)Co irradiation effect on cooked rice texture and amylopectin structure

The effect of γ irradiation on the cooked rice texture and amylopectin structure is studied with three varieties (Zhefu 504, Zhefu 802 and Zaogeng T2) subjected to 4kGy, skGy and 12kGy irradiation. The results indicate that irradiation has significant effect on the texture of the cooked rice; the hardness, cohesiveness,gumminess and chewiness reduce significantly with the increase of the irradiation dose.Irradiation also affects the chain length distribution of the amylopectin, the side-chains of amylopectin of Zhefu 504 with the 12DP, Zhefu 802 with 24DP13 and Zaogeng T2 with 24DP13 and 36DP25 are broken.

双波长光度法测定热处理橡胶木中支链淀粉和直链淀粉的含量

橡胶木中直链淀粉和支链淀粉的组成比例直接影响淀粉的化学特性,橡胶木热处理过程中,淀粉发生不同程度热降解影响其比例发生变化,分析其含量变化将为热处理橡胶木热解机制研究提供理论基础.研究采用单因素法优化了提取方法参数,并建立了双波长光度法检测橡胶木中直链淀粉和支链淀粉含量的定量分析方法,检验了该分析方法的精密度、稳定性和加标回收率.结果表明,直链淀粉和支链淀粉的标准曲线方程分别为y=0.02499x-0.1136和y=0.00159x-0.0005,其回归系数R^(2)均为0.998,方法前处理样品简便、快速,分析方法稳定性好、灵敏度高,回收率稳定,适用于橡胶木中直链淀粉和支链淀粉含量的定量分析.分析比较了橡胶木素材及不同温度(125、155、185℃)热处理橡胶木中直链淀粉和支链淀粉的含量,橡胶木素材中支链淀粉含量远高于直链淀粉,质量比约为8∶1.随着热处理温度升高至155℃,直链淀粉含量出现明显下降,而温度升高至185℃,支链淀粉含量出现明显下降趋势.

Effects of the activities of key enzymes involved in starch biosynthesis on the fine structure of amylopectin in developing rice (Oryza sativa L.) endosperms

The dynamic changes of the activities of enzymes involving in starch biosynthesis, including ADP-glucose pyrophosphorylase (AGPase), soluble starch synthases (SSS), starch branching enzyme (SBE) and starch debranching enzymes (DBE) were studied, and changes of fine structure of amy- lopectin were characterized by isoamylase treatment during rice grain development, using trans anti-waxy gene rice plants. The relationships between the activities of those key enzymes were also analyzed. The amylose synthesis was significantly inhibited in transgenic Wanjing 9522, but the total starch content and final grain weight were less affected as compared with those of non-transgenic Wanjing 9522 rice cultivar. Analyses on the changes of activities of enzymes involving in starch bio- synthesis showed that different enzyme activities were expressed differently during rice endosperm development. Soluble starch synthase is relatively highly expressed in earlier stage of endosperm de- velopment, whilst maximal expression of granule-bound starch synthase (GBSS) occurred in mid-stage of endosperm development. No obvious differences in changes of the activities of AGPase and SBE between two rice cultivars investigated, except the DBEs. Distribution patterns of branches of amy- lopectin changed continually during the development of rice grains and varied between two rice culti- vars. It was suggested that amylopectin synthesis be prior to the synthesis of amylose and different enzymes have different roles in controlling syntheses of branches of amylopectin.

应用CRISPR/Cas9基因编辑技术获得高直链淀粉水稻种质

【目的】通过CRISPR/Cas9基因编辑技术遗传改良水稻种质,创制高直链淀粉新材料。【方法】以水稻品种中花11为试验材料,利用CRISPR/Cas9编辑系统对水稻淀粉分支酶(Starch branching enzymes,SBE)基因OsSBE3进行靶向编辑,利用PCR技术鉴定无标记纯合突变体,并测定其淀粉含量。【结果】T_(0)代获得10株突变体株系,T1代获得5个无标记纯合突变株系,其中4个株系(sbe3-22-6、sbe3-25-3、sbe3-25-4、sbe3-25-6)的直链淀粉含量和淀粉直支比显著高于野生型。【结论】本研究创制了高直链淀粉含量的水稻新种质,为水稻品质改良提供了参考。

新创制豫南黑香糯米的营养特性分析

以培育新品系及加工为目的,对4份诱变创制的豫南黑香糯米的淀粉、花青素和矿物质进行测定。结果表明,创制的不同品种的黑香糯米淀粉含量、花青素以及矿物质含量均有明显的差异。通过电子显微镜扫描观察发现,黑香糯米的淀粉颗粒结构与‘NIP’相比并没有显著性差异,但黑香糯米中淀粉含量(36.5%~50.2%)、提取率(52.8%~60.7%)、提取纯度(70.0%~80.0%)均低于‘NIP’(64.7%、71.2%和90.0%以上)。在淀粉的组成上,黑香糯米‘LN48⁃2’和‘LN48⁃3’品系与‘NIP’相近,直链淀粉占14%~16%,支链淀粉占84%~86%,而‘LN54’和‘LN55’品系与‘LN56 CK’品种相似,淀粉中支链淀粉约占100%。通过超高效液相色谱分析,‘LN48⁃3’中花青素含量最高,为109.29 mg/100 g,‘LN48⁃2’中花青素含量最低,为84.39 mg/100 g,但其花青素组成相同,均由矢车菊素⁃3⁃O⁃葡萄糖苷和芍药素⁃3⁃O⁃葡萄糖苷组成。采用等离子质谱仪分析黑香糯米中矿物质含量,‘LN48⁃2’品系的钙元素和锌元素含量最高,分别为1370.8 mg/kg和60.3 mg/kg,其次是‘LN54’,分别为956.3 mg/kg和55.2 mg/kg;新创制5个品系的铁元素含量变幅范围为57.0~82.3 mg/kg,高于对照黑香糯米‘LN56 CK’。

用双波长分光光度法测定鲜食玉米中直链淀粉和支链淀粉含量

以碘为显色剂,应用双波长分光光度法测定了鲜食玉米中直链淀粉和支链淀粉的含量.根据碘-直链淀粉和碘-支链淀粉复合物的吸收光谱,选择直链淀粉的测定波长为629 nm和463 nm,支链淀粉的测定波长为553 nm和738 nm,并依据回归方程求出直链淀粉和支链淀粉的含量.直链淀粉的含量在10～60 mg/L、支链淀粉在60～200 mg/L范围内符合比耳定律.用双波长法测定不同品种鲜食玉米中直链淀粉和支链淀粉含量取得了良好的效果.

叶面喷施钼锌肥对冬小麦籽粒品质的影响

【目的】探明钼锌肥配施对冬小麦籽粒品质的影响,为钼锌肥的应用及冬小麦籽粒品质的改善提供参考。【方法】在河南省冬小麦产区开展叶面喷施钼锌试验,以喷施清水为CK,设置喷施0.10%钼肥、喷施0.10%锌肥、喷施0.10%钼肥+0.10%锌肥处理,考察不同处理小麦籽粒品质的变化。【结果】叶面单独施钼、单独施锌和钼锌配施能有效提高冬小麦籽粒中支链淀粉含量,在一定程度上能增加淀粉总量;钼锌配施能有效提高冬小麦籽粒中醇溶蛋白和麦谷蛋白含量。与CK相比,钼锌配施冬小麦籽粒中钼含量提高0.03~0.09 mg/kg,增幅为10.34%~39.13%;锌含量增加0.88~4.34 mg/kg,增幅为3.65%~18.11%。【结论】冬小麦叶面喷施钼肥与锌肥均能有效提高籽粒中支链淀粉含量,钼锌配施能有效提高籽粒中醇溶蛋白和麦谷蛋白含量,钼锌肥在优质冬小麦生产中具有较高的应用价值。

Comparison of selective flocculation of low grade goethitic iron ore fines using natural and synthetic polymers and a graft copolymer

This study aims to beneficiate low grade goethitic iron ore fines using a selective flocculation process. Selective flocculation studies were conducted using different polymers such as starch amylopectin(AP), poly acrylic acid(PAA), and a graft copolymer(AP-g-PAA). The obtained results were analyzed; they indicate the enhancement of the iron ore grade from 58.49% to 67.52% using AP-g-PAA with a recovery of 95.08%. In addition, 64.45% Fe with a recovery of 88.79% was obtained using AP. Similarly, using PAA, the grade increased to 63.46% Fe with a recovery of 82.10%. The findings are also supported by characterizing concentrates using X-ray diffraction(XRD) and electron probe microanalysis(EPMA) techniques.

南瓜淀粉组分高效检测体系建立及应用

旨在建立高效的南瓜淀粉组分检测技术,以期提高定向育种和合理开发南瓜新品种的效率。根据双波长比色原理,利用提纯的南瓜直链淀粉和支链淀粉,通过等吸收点作图法,确定南瓜淀粉组分测定波长和参比波长。通过单因素比较试验,确定双波长测定南瓜淀粉组分的200μL反应体系。最后比较不同类型的南瓜淀粉组分含量,筛选极端材料。结果表明,直链淀粉的测定波长为616 nm、参比波长为472 nm,南瓜支链淀粉的测定波长为545 nm、参比波长为725 nm。建立南瓜直链淀粉和支链淀粉回归方程,直链淀粉浓度在0.0125~1 mg/mL、支链淀粉浓度在0.025~2 mg/mL范围内,线性关系良好(R2>0.9900)。96孔板200μL反应体系为待测液20μL、醋酸盐缓冲反应液(pH 3.5)90μL、12倍碘液稀释液90μL。供试23份南瓜材料的直链淀粉含量在0.52~26.68 mg/g果肉鲜重,支链淀粉含量在0.03~76.13 mg/g果肉鲜重,淀粉总量在0.55~101.61 mg/g果肉鲜重。以10和15 mg/g果肉鲜重、35和45 mg/g果肉鲜重、55和65 mg/g果肉鲜重,可以分别初步评价供试南瓜直链淀粉、支链淀粉和总淀粉含量低、中、高的情况。中印南瓜自交系比供试中国南瓜和印度南瓜的淀粉组分含量均高,最终筛选到7个中印南瓜自交系,用以淀粉定向育种潜力大。该试验获得的南瓜淀粉组分高效检测体系,对提高南瓜淀粉育种效率,行之有效。

High amylose to amylopectin ratios in nitrogen-free diets decrease the ileal endogenous amino acid losses of broiler chickens

This study explored the variation of ileal endogenous amino acid(IEAA)losses and its influencing factors in chickens offered nitrogen-free diets(NFD)containing different ratios of amylose to amylopectin(AM/AP).A total of 252 broiler chickens at 28 d old were randomly allocated into 7 treatment groups for a 3-d trial.The dietary treatments included a basal diet(control),a NFD containing corn starch(CS),and 5NFD with AM/AP ratios of 0.20,0.40,0.60,0.80,and 1.00,respectively.As the AM/AP ratio increased,the IEAA losses of all AAs,starch digestibility and maltase activity linearly decreased(P<0.05),but the DM digestibility linearly and quadratically decreased(P<0.05).Compared with the control,the NFD increased the number of goblet cells and its regulatory genes mucin-2 and krüppel-like factor 4(KLF-4)while decreasing serum glucagon and thyroxine concentrations,ileal villus height,and crypt depth(P<0.05).Additionally,NFD with lower AM/AP ratios(0.20 and 0.40)decreased the ileal microbiota species richness(P<0.05).In all NFD groups,the number of Proteobacteria increased whereas the abundance of Firmicutes dropped(P<0.05).However,the broilers in the AM/AP 0.60 group were closer to the digestive physiological state of chickens fed the control diet,with no significant change in maltase activity and mucin-2 expression(P<0.05).In conclusion,increasing AM/AP ratio in a NFD decreased the IEAA losses and the apparent ileal digestibility of starch but inevitably resulted in malnutrition and disruption of gut microbiota homeostasis.This study recommends AM/AP in NFD at 0.60 to measure IEAA of broiler chickens.

储藏对稻米淀粉结构及含量的影响

综述了储藏对稻米淀粉的含量及其结构影响。分析了稻米淀粉的结构,讨论了储藏对稻米直链淀粉和支链淀粉含量与结构的影响。在此基础上,提出加强储藏过程中稻米中支链淀粉和直链淀粉中不溶性组分的含量及其结构变化等的研究,将有利于解析储藏过程中稻米淀粉特性的变化及其对稻米品质的影响。

Characterization of eating quality and starch properties of two Wx alleles japonica rice cultivars under different nitrogen treatments

To understand the effect of nitrogen(N) fertilizer on rice(Oryza sativa L.) eating and cooking quality(ECQ). Here, we investigated the ECQ attributes, physicochemical foundation of ECQ, and amylopectin fine structure of two Waxy(Wx) alleles japonica rice cultivars Nanjing 9108(NJ9108) and Huaidao 5(HD5) under four N rates(0, 150, 300, and 450 kg ha^-1). Sensory and pasting properties of the two cultivars varied depending on N rates. Compared with the control(0 kg ha^-1), the overall eating quality and sensory value were significantly decreased under the N rates of 300 and 450 kg ha^-1. Further, conventional descriptive analysis showed that the stickiness and retrogradation of cooked rice were significantly decreased. These results indicated that application of N fertilizer seems to affect the texture of cooked rice, causing it to be less sticky, lowering its retrogradation, and consequently reducing its palatability. Results from rapid visco analyzer(RVA) revealed that the peak and breakdown viscosities significantly decreased, while the setback viscosity and peak time increased under the N rates of 300 and 450 kg ha^-1. However, no significant difference was observed when the N rate was 150 kg ha^-1, indicating that less N fertilization can maintain rice ECQ. As the N rate increasing, protein content increased, whereas apparent amylose content, starch content, and gel consistency almost unchanged. Interestingly, compared with the control, under N treatments, the percentage of short amylopectin branches in NJ9108 was decreased, but increased in HD5, as controlled by amylopectin synthesis-related genes. Notably, SSI and BEIIb were down-regulated in NJ9108, whereas BEIIb was up-regulated in HD5. Thus, the palatability of both rice cultivars was significantly decreased under excessive N fertilization as a consequence of reduced stickiness and retrogradation of the cooked rice, which might have resulted from an elevated protein content and altered amylopectin fine structure. In addition, amylopectin synthesis appeared to be affected by N fertilizer and the genotype of the rice cultivar.

Optical Sum Frequency Generation Image of Rice Grains

We observed optical sum frequency generation (SFG) images of cross-sections of glutinous rice grains, in order to test a possibility of the SFG microscopy as a tool for monitoring polysaccharide species in rice grains. The SFG response in the CH vibration range was the most intense in the crush cell layer at the edge of the endosperm adjacent to the embryo probably due to optical reflection and scattering effectby the rugged dielectric structure of the crush cell layer. The SFG spectra as a function of the infrared wavelength depended on the measurement position in the endosperm. The SFG results were compared with those by Raman and infrared spectroscopies for the same samples.